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Improved Adenovirus Type 5 Vector-Mediated Transduction of Resistant Cells by Piggybacking on Coxsackie B-Adenovirus Receptor-Pseudotyped Baculovirus?  

PubMed Central

Taking advantage of the wide tropism of baculoviruses (BVs), we constructed a recombinant BV (BVCAR) pseudotyped with human coxsackie B-adenovirus receptor (CAR), the high-affinity attachment receptor for adenovirus type 5 (Ad5), and used the strategy of piggybacking Ad5-green fluorescent protein (Ad5GFP) vector on BVCAR to transduce various cells refractory to Ad5 infection. We found that transduction of all cells tested, including human primary cells and cancer cell lines, was significantly improved using the BVCAR-Ad5GFP biviral complex compared to that obtained with Ad5GFP or BVCARGFP alone. We determined the optimal conditions for the formation of the complex and found that a high level of BVCAR-Ad5GFP-mediated transduction occurred at relatively low adenovirus vector doses, compared with transduction by Ad5GFP alone. The increase in transduction was dependent on the direct coupling of BVCAR to Ad5GFP via CAR-fiber knob interaction, and the cell attachment of the BVCAR-Ad5GFP complex was mediated by the baculoviral envelope glycoprotein gp64. Analysis of the virus-cell binding reaction indicated that the presence of BVCAR in the complex provided kinetic benefits to Ad5GFP compared to the effects with Ad5GFP alone. The endocytic pathway of BVCAR-Ad5GFP did not require Ad5 penton base RGD-integrin interaction. Biodistribution of BVCAR-Ad5Luc complex in vivo was studied by intravenous administration to nude BALB/c mice and compared to Ad5Luc injected alone. No significant difference in viscerotropism was found between the two inocula, and the liver remained the preferred localization. In vitro, coagulation factor X drastically increased the Ad5GFP-mediated transduction of CAR-negative cells but had no effect on the efficiency of transduction by the BVCAR-Ad5GFP complex. Various situations in vitro or ex vivo in which our BVCAR-Ad5 duo could be advantageously used as gene transfer biviral vector are discussed.

Granio, Ophelia; Porcherot, Marine; Corjon, Stephanie; Kitidee, Kuntida; Henning, Petra; Eljaafari, Assia; Cimarelli, Andrea; Lindholm, Leif; Miossec, Pierre; Boulanger, Pierre; Hong, Saw-See



Coxsackie viral myocarditis  

PubMed Central

Coxsackie viral myocarditis is a common disease, yet idiopathic dilated cardiomyopathy is a less common consequence. Insights gained from studying the Coxsackie virus B-3 murine model of myocarditis has led to the hypothesis that an acute Coxsackie viral myocarditis can result in persistent, non-viral mediated cellular responses that result in a chronic inflammatory state leading to progressive myocyte loss and ultimate development of dilated cardiomyopathy. Although the evidence linking myocarditis to dilated cardiomyopathy is circumstantial in man, the identification of defects in immunoregulation may provide the impetus to further research into the pathogenesis and ultimately the development of more rational therapies directed at modulating immune responses to alter the natural history of clinical dilated cardiomyopathy.

O'Connell, John B.; Robinson, John A.



Myocardial imaging. Coxsackie myocarditis  

SciTech Connect

A 3-week-old male neonate with heart failure associated with Coxsackie virus infection was imaged with Tc-99m PYP and TI-201. The abnormal imaging pattern suggested myocardial infarction. Autopsy findings indicated that the cause was myocardial necrosis secondary to an acute inflammatory process. Causes of abnormal myocardial uptake of Tc-99m PYP in pediatrics include infarction, myocarditis, cardiomyopathy, bacterial endocarditis, and trauma. Myocardial imaging cannot provide a specific cause diagnosis. Causes of myocardial infarction in pediatrics are listed in Table 1.

Wells, R.G.; Ruskin, J.A.; Sty, J.R.



Coxsackie A10 Virus Infection Among Infectious Hepatitis Contacts  

PubMed Central

During an outbreak of infectious hepatitis at a housing development, Coxsackie A10 virus was recovered from the stools of 45 different contacts and from the blood of four others. Caution should be exercised in attributing an etiological role to any given isolate of a Group A Coxsackie virus in view of the widespread distribution of these organisms. Nevertheless, the recovery of Coxsackie A10 viruses from the blood and stools of contacts with hepatitis cases appears to warrant record.

Embil, J. A.; Van Rooyen, C. E.; Nagler, F. P.



Coxsackie B viruses and heart muscle disease in Nigerian adults.  


In 44 Nigerians with heart muscle disease, the percentage of patients who had a fourfold rise or fall to at least one of the Coxsackie B viruses was greater than controls. Patients with heart muscle disease also had significantly higher levels of antibody titres. Antibodies to Coxsackie viruses B2, B3 and B6 were more frequent in their sera than in controls. While these results do not prove a Coxsackie B viral aetiology it is concluded that these viruses could be the cause of cardiac damage and heart failure in some Nigerians. It could also be one of the many adverse factors which produce repeated myocardial damage and thus weaken the heart to the point of failure. PMID:228451

Falase, A O; Fabiyi, A; Odegbo-Olukoya, O O



Neonate with coxsackie B1 infection, cardiomyopathy and arrhythmias.  

PubMed Central

A neonate with Coxsackie B1 infection who experienced significant congestive heart failure, cardiomyopathy and arrhythmia is reported. Viral myocarditis, an important cause of acquired heart disease in neonates, should be considered in the differential diagnosis of neonatal congestive heart failure and cardiomyopathy. A review of the literature is presented.

Lu, Jimmy C.; Koay, Kelly W.; Ramers, Christian B.; Milazzo, Angelo S.



Reduced coxsackie antibody titres in Type 1 (insulin-dependent) diabetic patients presenting during an outbreak of coxsackie B 3 and B 4 infection  

Microsoft Academic Search

Summary  The potential role of antecedent viral infection in the pathogenesis of Type 1 (insulin-dependent) diabetes was investigated by measuring antibody titres to several viruses in serum obtained at the time of diagnosis of diabetes. An outbreak of Coxsackie B 4 infection folio wed by a wave of Coxsackie B 3 and B 5 infections occurred in Seattle during the time

J. P. Palmer; M. K. Cooney; R. H. Ward; J. A. Hansen; J. B. Brodsky; C. G. Ray; J. R. Crossley; C. M. Asplin; R. H. Williams



Epidemic Coxsackie B virus infection in Johannesburg, South Africa.  


A particularly extensive epidemic of Coxsackie B3 virus infection occurred in Johannesburg in the spring and summer of 1984. A total of 142 positive cases were diagnosed by isolation of the virus from stools and other specimens (60) or by serology (82). Coxsackie B3 accounted for 87% of the isolations and was also the dominant serotype on serology. The outbreak involved predominantly children and young adults, with no apparent sex differences being noted. The majority of specimens came from the white population and no significant difference in age or sex distribution could be observed between the two race groups. The major clinical presentation in the white group was Bornholm disease followed by cardiac involvement and then meningoencephalitis. In the black group, however, myocarditis was the major clinical presentation, which is of particular interest taking into account the extremely high incidence of acute rheumatic carditis in this population and the prevalence of chronic cardiomyopathy. PMID:2999227

Schoub, B D; Johnson, S; McAnerney, J M; Dos Santos, I L; Klaassen, K I



Hepatitis and Encephalitis due to Coxsackie Virus A9 in an Adult  

PubMed Central

Coxsackie virus infection most commonly manifests itself in the neonatal period as a multisystem disease. This life-threatening neonatal infection has been recently treated with a new anti-picornaviral drug, pleconaril. In contrast, in adults Coxsackie virus is an uncommon source of hepatitis, but Coxsackie virus type B has been described in case reports to cause hepatitis. This is the first case report of hepatitis and encephalitis secondary to Coxsackie virus type A9 in an adult. This virus was found in a culture of the cerebrospinal fluid and was confirmed by PCR. The patient recovered completely without specific treatment.

Moreau, Brigitte; Bastedo, Clare; Michel, Rene P.; Ghali, Peter



Structural variations in species B adenovirus fibers impact CD46 association.  


A majority of species B adenoviruses (Ads) use CD46 as their primary receptor; however, the precise mechanisms involved in the binding of different Ad types to CD46 have not been resolved. Although previous studies indicate close similarities between two members of species B2 Ads in their usage of CD46, our current investigations revealed a surprisingly low CD46 binding affinity of the species B1 Ad16 fiber knob (equilibrium dissociation constant of 437 nM). We determined the crystal structure of the Ad16 fiber knob and constructed a model of this protein in complex with CD46. A comparison of this model to that of the CD46-Ad11 complex revealed structural differences in the FG and IJ loops that are part of the CD46 binding site. An analysis of a panel of recombinant fiber knobs with mutations targeting these regions in Ad16 and Ad11 uncovered a major contribution of the FG loop on CD46 binding. Two extra residues in the FG loop of the Ad16 fiber significantly reduce receptor interaction. Although avidity effects permit the use of CD46 on host cells by Ad16, virus binding occurs with lower efficiency than with B2 Ad types. The longer FG loop of the Ad16 fiber knob also is shared by other species B1 Ad fibers and, thus, may contribute to the low CD46 binding efficiencies observed for these Ad types. Our findings provide a better understanding of how different Ad types associate with CD46 and could aid in the selection of specific Ad fibers for more efficient Ad gene delivery vectors. PMID:18524830

Pache, Lars; Venkataraman, Sangita; Reddy, Vijay S; Nemerow, Glen R



Structural Variations in Species B Adenovirus Fibers Impact CD46 Association  

SciTech Connect

A majority of species B adenoviruses (Ads) use CD46 as their primary receptor; however, the precise mechanisms involved in the binding of different Ad types to CD46 have not been resolved. Although previous studies indicate close similarities between two members of species B2 Ads in their usage of CD46, our current investigations revealed a surprisingly low CD46 binding affinity of the species B1 Ad16 fiber knob (equilibrium dissociation constant of 437 nM). We determined the crystal structure of the Ad16 fiber knob and constructed a model of this protein in complex with CD46. A comparison of this model to that of the CD46-Ad11 complex revealed structural differences in the FG and IJ loops that are part of the CD46 binding site. An analysis of a panel of recombinant fiber knobs with mutations targeting these regions in Ad16 and Ad11 uncovered a major contribution of the FG loop on CD46 binding. Two extra residues in the FG loop of the Ad16 fiber significantly reduce receptor interaction. Although avidity effects permit the use of CD46 on host cells by Ad16, virus binding occurs with lower efficiency than with B2 Ad types. The longer FG loop of the Ad16 fiber knob also is shared by other species B1 Ad fibers and, thus, may contribute to the low CD46 binding efficiencies observed for these Ad types. Our findings provide a better understanding of how different Ad types associate with CD46 and could aid in the selection of specific Ad fibers for more efficient Ad gene delivery vectors.

Pache, Lars; Venkataraman, Sangita; Reddy, Vijay S.; Nemerow, Glen R. (Scripps)



COXSACKIE VIRUSES--A Review of Pathologic, Epidemiologic, Diagnostic and Etiologic Observations  

PubMed Central

Coxsackie disease comprises three clinical entities—herpangina, so-called non-paralytic poliomyelitis, and epidemic pleurodynia. Several strains of antigenically-related viruses, Groups A and B, designated as Coxsackie virus have been isolated from stool specimens and from material from the throat of many patients with the diseases mentioned. Inasmuch as the virus has also been recovered from normal persons, there is as yet uncertainty as to causal relationship between the presence of the virus and the disease. Reports of the isolation of Coxsackie virus and poliomyelitis virus from the same patient make difficult the interpretation of the findings. The diagnosis of Coxsackie disease entails animal inoculation and serologic procedures. Emphasis is placed on the necessity of obtaining stool specimens, throat washings, and “paired” blood specimens from patients suspected of the disease.

Carpenter, Charles M.; Boak, Ruth A.



COXSACKIE VIRUS IN SOUTHERN CALIFORNIA--Isolation of a Strain from Stools of a Patient  

PubMed Central

Thirty-three stool specimens from 29 patients were examined for Coxsackie virus by the inoculation of suckling mice. Such a virus, designated “California I,” was obtained from two stool specimens collected on successive days from a patient with so-called nonparalytic poliomyelitis. Neutralizing antibodies for the California I strain of Coxsackie virus could not be demonstrated in serum obtained from the patient early in the illness, but were present in convalescent serum. Serum from the patient's daughter, who previously had had a similar illness, neutralized the strain of virus isolated from the father. In pathologic examination of the skeletal muscles of mice infected with the California I virus, lesions typical of those produced by Coxsackie virus, group A, were noted. California I strain of the virus was not neutralized by immune serum prepared from several other strains of Coxsackie virus.

Boak, Ruth A.; Klaumann, Benjamin F.; Ward, C. Bradley



The role of T lymphocytes in the pathogenesis of Coxsackie virus B3 heart disease.  

PubMed Central

Myocarditis in athymic BALB/c-nu/nu (nu/nu) mice infected with Coxsackie virus B3 was studied to determine whether inflammatory mononuclear cell infiltration was produced by transfer of spleen cells ob BALB/c-nu/+ (nu/+) mice infected with the same virus. In addition, spleen cells of uninfected nu/+mice were transferred into athymic nu/nu mice infected with Coxsackie virus B3. Athymic nu/nu mice infected with Coxsackie virus B3 after transfer of spleen cells of nu/+ mice infected with the same virus developed marked to moderate myocarditis with inflammatory mononuclear cell infiltration. However, athymic nu/nu mice infected with Coxsackie virus B3 after transfer of spleen cells of uninfected nu/+ mice developed moderate to mild degeneration or necrosis of the muscle fibres, although mild mononuclear cell infiltration was found in only one mouse. The incidence of myocardial lesions of athymic nu/nu mice infected with Coxsackie virus B3 after transfer of infected-spleen cells of nu/+ mice was about the same is that in infected nu/+ mice after transfer of spleen cells of uninfected nu/+ mice. However, degrees in the intensity of the muscular changes and inflammatory mononuclear cell infiltration in the former was significantly greater than that in the latter. The results show, therefore, that Coxsackie virus B3 infection stimulated production of cytotoxic activity in the spleen which was evident on Day 6. From the present results, it is confirmed that myocarditis in mice infected with Coxsackie virus B3 is thymus-dependent, supporting previous investigations. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5

Hashimoto, I.; Tatsumi, M.; Nakagawa, M.



Coxsackie-B-virus-specific IgM responses in patients with cardiac and other diseases.  


An enzyme-linked immunosorbent assay (ELISA) test using polyvalent antigens and antisera was developed to detect Coxsackie-B-virus-specific IgM responses. The sera of 24 of 64 (37.5%) patients with acute pericarditis and 14 of 38 (36%) with acute myocarditis were positive for Coxsackie-B-virus-specific IgM. 4 of 30 (13.3%) patients with acute ischaemic heart disease and 2 of 28 (7.1%) patients with congestive cardiomyopathy were also positive. Coxsackie-B-virus-specific IgM was detected in the sera of 21 of 57 (36.8%) patients with Bornholm disease and 2 of 4 patients with acute-onset juvenile diabetes. Coxsackie-B-virus-specific IgM responses persisted for 6-8 weeks. Sera from patients with chronic valvular heart disease, Mycoplasma pneumoniae infections, and virus infections caused by viruses other than Coxsackie-B viruses were all negative. False-positive results did not occur when sera containing high titres of rheumatoid factor were tested. PMID:6107769

El-Hagrassy, M M; Banatvala, J E; Coltart, D J



Susceptibility of Skeletal Muscle to Coxsackie A2 Virus Infection: Effects of Botulinum Toxin and Denervation  

NASA Astrophysics Data System (ADS)

Coxsackie A viruses can infect denervated but not innervated mature skeletal muscles. The role of synaptic transmission in preventing susceptibility to Coxsackievirus infection was studied by surgically denervating leg muscles of mice or injecting the muscles with botulinum toxin to block quantal release of acetylcholine. Control muscles were injected with heat-inactivated toxin. Subsequent injection of Coxsackie A2 virus resulted in extensive virus replication and tissue destruction in the denervated and botulinum toxin-treated muscles, while the control muscles showed only minimal changes. This suggests that the susceptibility of skeletal muscle to Coxsackievirus infection is regulated by synaptic transmission.

Andrew, Clifford G.; Drachman, Daniel B.; Pestronk, Alan; Narayan, Opendra



Acanthamoeba castellanii Promotion of In Vitro Survival and Transmission of Coxsackie B3 Viruses  

Microsoft Academic Search

This work was undertaken to determine whether Acanthamoeba could play a role in the survival and transmission of coxsackieviruses and focused on in vitro interactions between Acanthamoeba castellanii and coxsackie B3 viruses (CVB-3). Residual virus titer evaluations and immunofluorescence experiments revealed a remarkable CVB-3 adsorption on amoeba surfaces and accumulation inside cells. The survival of viruses was independent of the

A. Mattana; C. Serra; E. Mariotti; G. Delogu; P. L. Fiori; P. Cappuccinelli



A recent epidemic of Coxsackie virus type A24 acute haemorrhagic conjunctivitis in Singapore  

Microsoft Academic Search

A recent epidemic of acute conjunctivitis in Singapore showed again the importance of Coxsackie virus type A24 variant as a causative agent of acute haemorrhagic conjunctivitis (AHC). Although the ocular manifestations appeared similar to those described for the 1970 and 1975 outbreaks, a markedly higher rate of respiratory involvements was noted. Not observed in previous epidemics were herpes-like vesicles in

M Yin-Murphy; Baharuddin-Ishak; M C Phoon; V T Chow



Isoform-Specific Regulation and Localization of the Coxsackie and Adenovirus Receptor in Human Airway Epithelia  

Microsoft Academic Search

Adenovirus is an important respiratory pathogen. Adenovirus fiber from most serotypes co-opts the Coxsackie-Adenovirus Receptor (CAR) to bind and enter cells. However, CAR is a cell adhesion molecule localized on the basolateral membrane of polarized epithelia. Separation from the lumen of the airways by tight junctions renders airway epithelia resistant to inhaled adenovirus infection. Although a role for CAR in

Katherine J. D. A. Excoffon; Nicholas D. Gansemer; Matthew E. Mobily; Philip H. Karp; Kalpaj R. Parekh; Joseph Zabner



[Acute severe coxsackie virus B myocarditis of pseudonecrotic form. Apropos of 2 cases].  


This study reports two cases of acute severe Coxsackie virus B4 myocarditis in which the immediate clinical signs suggested the acute phase of myocardial infarction, apparently antero-lateral in the first case in a context of cardiogenic shock and infero-lateral in the second case, in the context of acute pulmonary edema. Both cases were characterized by the severity of the initial signs. Numerous other cases of acute Coxsackie virus B myocarditis, simulating myocardial infarction, have been reported in the literature and these contexts deserve to be recognized earlier as they call for specific treatment. The immediate outcome was favorable in both cases but required massive cardiological intensive care in the first patient. Long term follow-up was excellent. PMID:8396381

Beard, T; Boudjemaa, B; Carrié, D; Chakra, G; Ferrières, J; Delay, M; Bernadet, P



Phyllaemblicin B inhibits Coxsackie virus B3 induced apoptosis and myocarditis  

Microsoft Academic Search

Coxsackie virus B3 (CVB3) is believed to be a major contributor to viral myocarditis since virus-associated apoptosis plays a role in the pathogenesis of experimental myocarditis. In this study, we investigated the in vitro and in vivo antiviral activities of Phyllaemblicin B, the main ellagitannin compound isolated from Phyllanthus emblica, a Chinese herb medicine, against CVB3. Herein we report that

Ya-Feng Wang; Xiao-Yan Wang; Zhe Ren; Chui-Wen Qian; Yi-Cheng Li; Kitazato Kaio; Qing-Duan Wang; Yan Zhang; Li-Yun Zheng; Jin-Hua Jiang; Chong-Ren Yang; Qing Liu; Ying-Jun Zhang; Yi-Fei Wang



A recent epidemic of Coxsackie virus type A24 acute haemorrhagic conjunctivitis in Singapore.  


A recent epidemic of acute conjunctivitis in Singapore showed again the importance of Coxsackie virus type A24 variant as a causative agent of acute haemorrhagic conjunctivitis (AHC). Although the ocular manifestations appeared similar to those described for the 1970 and 1975 outbreaks, a markedly higher rate of respiratory involvements was noted. Not observed in previous epidemics were herpes-like vesicles in the conjunctiva and eyelids of one patient and vesicles in the buccal mucosa and lips of another from whom Coxsackie virus A24 was isolated. The most interesting finding in this study was the isolation of five wild (non-Sabin) poliovirus type 1 strains. Three strains were obtained from conjunctival and two from throat swabs of patients with mild to severe conjunctivitis. It is conceivable that the rare reports of polio-like paralysis or radiculomyelitis accompanying or following AHC in a few Asian countries could be attributed to concurrent infections with a poliovirus and either enterovirus type 70 or Coxsackie virus type A24. PMID:3024697

Yin-Murphy, M; Baharuddin-Ishak; Phoon, M C; Chow, V T



A case of fulminant type 1 diabetes with coxsackie B4 virus infection diagnosed by elevated serum levels of neutralizing antibody.  


A 39-year-old woman with hyperglycemia and ketonuria but with normal HbA1c level was diagnosed as having fulminant type 1 diabetes. The patient had 8-fold increase in the plasma titer of coxsackie B4 virus neutralizing antibody. Infection with coxsackie B4 virus was associated with fulminant type 1 diabetes. PMID:19362384

Akatsuka, Hajime; Yano, Yutaka; Gabazza, Esteban C; Morser, John; Sasaki, Ryoma; Suzuki, Toshinari; Fujiwara, Ryoko; Katsuki, Akira; Takei, Yoshiyuki; Sumida, Yasuhiro



Acute sensory and autonomic neuropathy: possible association with coxsackie B virus infection.  

PubMed Central

This report describes a 26 year old woman with a Coxsackie B virus infection complicated by an acute pandysautonomic and sensory neuropathy. Electrophysiological studies suggested an axonal neuropathy. A sural nerve biopsy performed early in the disease showed axonal degeneration with a virtual absence of unmyelinated fibres and moderate loss of myelinated fibres, mainly affecting the small fibres; this differs from previous reports. An immune-mediated or direct virus action might explain the pathogenesis of this unusual evolution of a viral infection. Images

Pavesi, G; Gemignani, F; Macaluso, G M; Ventrua, P; Magnani, G; Fiocchi, A; Medici, D; Marbini, A; Mancia, D



Acanthamoeba castellanii Promotion of In Vitro Survival and Transmission of Coxsackie B3 Viruses  

PubMed Central

This work was undertaken to determine whether Acanthamoeba could play a role in the survival and transmission of coxsackieviruses and focused on in vitro interactions between Acanthamoeba castellanii and coxsackie B3 viruses (CVB-3). Residual virus titer evaluations and immunofluorescence experiments revealed a remarkable CVB-3 adsorption on amoeba surfaces and accumulation inside cells. The survival of viruses was independent of the dynamics of amoeba replication and encystment. In addition, our results indicated that virus-infected amoebas can release infectious viruses during interaction with human macrophages. On the basis of these data, Acanthamoeba appears to be a potential promoter of the survival of coxsackieviruses and their transmission to human hosts.

Mattana, A.; Serra, C.; Mariotti, E.; Delogu, G.; Fiori, P. L.; Cappuccinelli, P.



Loss of the coxsackie and adenovirus receptor contributes to gastric cancer progression  

PubMed Central

Loss of the coxsackie and adenovirus receptor (CAR) has previously been observed in gastric cancer. The role of CAR in gastric cancer pathobiology, however, is unclear. We therefore analysed CAR in 196 R0-resected gastric adenocarcinomas and non-cancerous gastric mucosa samples using immunohistochemistry and immunofluorescence. Coxsackie and adenovirus receptor was found at the surface and foveolar epithelium of all non-neoplastic gastric mucosa samples (n=175), whereas only 56% of gastric cancer specimens showed CAR positivity (P<0.0001). Loss of CAR correlated significantly with decreased differentiation, increased infiltrative depths, presence of distant metastases, and was also associated with reduced carcinoma-specific survival. To clarify whether CAR impacts the tumorbiologic properties of gastric cancer, we subsequently determined the role of CAR in proliferation, migration, and invasion of gastric cancer cell lines by application of specific CAR siRNA or ectopic expression of a human full-length CAR cDNA. These experiments showed that RNAi-mediated CAR knock down resulted in increased proliferation, migration, and invasion of gastric cancer cell lines, whereas enforced ectopic CAR expression led to opposite effects. We conclude that the association of reduced presence of CAR in more severe disease states, together with our findings in gastric cancer cell lines, suggests that CAR functionally contributes to gastric cancer pathogenesis, showing features of a tumour suppressor.

Anders, M; Vieth, M; Rocken, C; Ebert, M; Pross, M; Gretschel, S; Schlag, P M; Wiedenmann, B; Kemmner, W; Hocker, M



Mycophenolate mofetil inhibits the development of Coxsackie B3-virus-induced myocarditis in mice  

PubMed Central

Background Viral replication as well as an immunopathological component are assumed to be involved in the development of coxsackie B virus (CBV)-induced myocarditis. We observed that mycophenolic acid (MPA), the active metabolite of the immunosuppressive agent mycophenolate mofetil (MMF), inhibits coxsackie B3 virus (CBV3) replication in primary Human myocardial fibroblasts. We therefore studied whether MMF, which is thus endowed with a direct antiviral as well as immunosuppressive effect, may prevent CBV-induced myocarditis in a murine model. Results Four week old C3H-mice were infected with CBV3 and received twice daily, for 7 consecutive days (from one day before to 5 days post-virus inoculation) treatment with MMF via oral gavage. Treatment with MMF resulted in a significant reduction in the development of CBV-induced myocarditis as assessed by morphometric analysis, i.e. 78% reduction when MMF was administered at 300 mg/kg/day (p < 0.001), 65% reduction at 200 mg/kg/day (p < 0.001), and 52% reduction at 100 mg/kg/day (p = 0.001). The beneficial effect could not be ascribed to inhibition of viral replication since titers of infectious virus and viral RNA in heart tissue were increased in MMF-treated animals as compared to untreated animals. Conclusion The immunosuppressive agent MMF results in an important reduction of CBV3-induced myocarditis in a murine model.

Padalko, Elizaveta; Verbeken, Erik; Matthys, Patrick; Aerts, Joeri L; De Clercq, Erik; Neyts, Johan




PubMed Central

Virus particles derived from single cells infected with two enteroviruses have been studied. Evidence was obtained to indicate that phenotypic, but not genotypic, mixing occurs between Coxsackie A9 (CAP) and ECHO 7 (E7) viruses. Monkey kidney cultures in monolayer were doubly infected with high multiplicities of CA9 and E7 viruses. During the latent period, the infected cells were suspended, diluted, and distributed under oil into droplets each containing a single cell, as checked by microscopic observation. The virus particles released by individual cells into the microdrop were characterized in differential plaque neutralization tests. Fifteen per cent of the microdrops contained doubly neutralizable particles, 53 per cent yielded either CA9 or E7 particles, and 34 yielded particles of an intermediate character (deficits between 37 and 75 per cent). On passage, the doubly neutralizable particles yielded progeny of both parental types. All passage strains behaved like the corresponding parent strain as regards pathogenicity for newborn mice, which is to say that this property was limited to virus particles with CA9 antigenicity. Coxsackie A9 has a more rapid growth cycle than ECHO 7 in rhesus monkey kidney cell cultures, and a slower one in patas cultures. In rhesus, when E7 virus was added first, CA9 could be added up to 2 hours later, and still a significant number of cells yielded either CA9 or doubly neutralizable virus. The converse was observed in patas cells.

Itoh, Heihachi; Melnick, Joseph L.



Acquisition of Antibodies to Various Coxsackie and Echo Viruses and Hepatitis A Virus in Agricultural Communal Settlements in Israel.  

National Technical Information Service (NTIS)

A seroepidemiological study was conducted to measure the antibody prevalence for eight different enteric viruses. These include seven 'classical' enteroviruses, ie, Coxsackie virus types A9, B1, B3, B4 and three ECHO virus types 4,7, and 9, as well as hep...

A. Morag B. Fattal H. I. Shuval M. Margalith



Essential Role of the Coxsackie - and Adenovirus Receptor (CAR) in Development of the Lymphatic System in Mice  

Microsoft Academic Search

The coxsackie- and adenovirus receptor (CAR) is a cell adhesion molecule predominantly associated with epithelial tight junctions in adult tissues. CAR is also expressed in cardiomyocytes and essential for heart development up to embryonic day 11.5, but not thereafter. CAR is not expressed in vascular endothelial cells but was recently detected in neonatal lymphatic vessels, suggesting that CAR could play

Momina Mirza; Mei-Fong Pang; Mohamad Amr Zaini; Paula Haiko; Tuomas Tammela; Kari Alitalo; Lennart Philipson; Jonas Fuxe; Kerstin Sollerbrant




EPA Science Inventory

The objectives of this research are to improve on the current analytical methods for quantitative detection of infective coxsackie and echo viruses in drinking water. The specific objectives of this research are to: (1) Improve the sensitivity and specificity of IMS-PCR for in...



EPA Science Inventory

A seroepidemiological study was conducted to measure the antibody prevalence for eight different enteric viruses. These include seven 'classical' enteroviruses, ie, Coxsackie virus types A9, B1, B3, B4 and three ECHO virus types 4,7, and 9, as well as hepatitis A virus (HAV), rec...


Molecular Classification of Coxsackie A Viruses on the Basis of the 5'-UTR: Structural and Evolutionary Aspects  

Microsoft Academic Search

  The sequences from a large part of the 5'-UTR of 21 coxsackie A virus (CAV) reference strains for which such data did not\\u000a exist in the past were obtained. Those sequences, along with the respective available sequences from the rest of the CAV reference\\u000a strains and many other enteroviruses, were compared. According to the results of this comparison, enteroviruses are

Nikolaos Siafakas; Panayotis Markoulatos; Glyn Stanway



The role of Coxsackie B viruses in the pathogenesis of myocarditis, dilated cardiomyopathy and inflammatory muscle disease.  


Coxsackie B viruses are members of the family Picornaviridae which have been associated by retrospective serology with a range of muscle diseases, particularly myocarditis, dilated cardiomyopathy and epidemic pleurodynia (epidemic myalgia or Bornholm disease). It has been proposed that virus-induced myocarditis disposes to the development of idiopathic dilated cardiomyopathy. However, despite many attempts, isolation of infectious virus or immunofluorescent detection of virus-specific antigens in the affected tissue is rare, although virus may be found in faeces early in infection. This discrepancy awaited the development of nucleic acid probes to resolve the problem of whether virus was present consistently in myocardium or other muscle tissues. We report here the synthesis of Coxsackie B virus-specific complementary DNA (cDNA) probes and their use in molecular hybridizations to quantitative slot-blots of RNA prepared from either endomyocardial or skeletal muscle biopsy specimens. Of 50 patients with histologically proven myocarditis or dilated cardiomyopathy, 28 (56%) had an endomyocardial biopsy specimen positive for the presence of Coxsackie B virus-specific RNA. Twenty-two patients with other cardiac diseases of known aetiology, unrelated to virus infection, were all negative. Multiple biopsies were obtained from 20 patients with myocarditis or dilated cardiomyopathy and 15 of these (75%) had at least one biopsy specimen positive, indicating the focal nature of the disease. In analogous investigations, Coxsackie B virus-specific RNA was detected in four out of seven single skeletal muscle biopsy specimens from patients suffering from juvenile dermatomyositis, and one out of two patients with adult polymyositis. Ten muscle controls, either normal or Duchenne muscular dystrophy, were negative for virus RNA. PMID:2847741

Archard, L C; Richardson, P J; Olsen, E G; Dubowitz, V; Sewry, C; Bowles, N E



Coxsackie and adenovirus receptor is a target and a mediator of estrogen action in breast cancer.  


The involvement of the coxsackie and adenovirus receptor (CAR), an adhesion molecule known to be the main determinant of adenovirus transduction of the cells, in cancer is currently under investigation. Recent reports suggest that CAR levels are elevated in breast cancer, and this may have an impact on its use as means of delivery for gene therapy. In this study, we show that estradiol (E(2)) treatment of the estrogen receptor (ER)-positive breast cancer cell MCF-7 increases CAR levels and, in turn, enhances adenoviral transduction. Employing the transfection of CAR promoters in breast cancer cells, we show that this regulation of CAR expression occurs at the transcriptional level. In addition, and by chromatin immunoprecipitation, we have identified a crucial region of CAR promoter that controls E(2) responsiveness of CAR gene through the recruitment of ER. Moreover, utilizing CAR antibodies or CAR silencing by RNA interference repressed the estrogen-dependent growth of breast cancer cells, whereas the stable expression of CAR in MCF-7 or MDA-MB-231 cells led to an increased proliferation. Altogether, our data suggest that CAR is a novel estrogen-responsive gene, which is involved in the E(2)-dependent proliferation of breast cancer cells. PMID:21389059

Vindrieux, David; Le Corre, Ludovic; Hsieh, Jer-Tsong; Métivier, Raphaël; Escobar, Pauline; Caicedo, Andrès; Brigitte, Madly; Lazennec, Gwendal



Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles  

PubMed Central

Background Coxsackie virus A16 (CVA16) infections have become a serious public health problem in the Asia-Pacific region. It manifests most often in childhood exanthema, commonly known as hand-foot-and-mouth disease (HFMD). There are currently no vaccine or effective medical treatments available. Principal Finding In this study, we describe the production, purification and characterization of CVA16 virus produced from Vero cells grown on 5 g/L Cytodex 1 microcarrier beads in a five-liter serum-free bioreactor system. The viral titer was found to be >106 the tissue culture's infectious dose (TCID50) per mL within 7 days post-infection when a multiplicity of infection (MOI) of 10?5 was used for initial infection. Two CVA16 virus fractions were separated and detected when the harvested CVA16 viral concentrate was purified by a sucrose gradient zonal ultracentrifugation. The viral particles detected in the 24–28% sucrose fractions had low viral infectivity and RNA content. The viral particles obtained from 35–38% sucrose fractions were found to have high viral infectivity and RNA content, and composed of four viral proteins (VP1, VP2, VP3 and VP4), as shown by SDS-PAGE analyses. These two virus fractions were formalin-inactivated and only the infectious particle fraction was found to be capable of inducing CVA16-specific neutralizing antibody responses in both mouse and rabbit immunogenicity studies. But these antisera failed to neutralize enterovirus 71. In addition, rabbit antisera did not react with any peptides derived from CVA16 capsid proteins. Mouse antisera recognized a single linear immunodominant epitope of VP3 corresponding to residues 176–190. Conclusion These results provide important information for cell-based CVA16 vaccine development. To eliminate HFMD, a bivalent EV71/CVA16 vaccine formulation is necessary.

Chong, Pele; Guo, Meng-Shin; Lin, Fion Hsiao-Yu; Hsiao, Kuang-Nan; Weng, Shu-Yang; Chou, Ai-Hsiang; Wang, Jen-Ren; Hsieh, Shih-Yang; Su, Ih-Jen; Liu, Chia-Chyi



Efficient adenovirus-mediated gene transfer into primary T cells and thymocytes in a new coxsackie\\/adenovirus receptor transgenic model  

Microsoft Academic Search

BACKGROUND: Gene transfer studies in primary T cells have suffered from the limitations of conventional viral transduction or transfection techniques. Replication-defective adenoviral vectors are an attractive alternative for gene delivery. However, naive lymphocytes are not readily susceptible to infection with adenoviruses due to insufficient expression of the coxsackie\\/adenovirus receptor. RESULTS: To render T cells susceptible to adenoviral gene transfer, we

Vincent Hurez; Robin Dzialo-Hatton; James Oliver; R James Matthews; Casey T Weaver



Absence of hepatic uptake of Tc-99m sulfur colloid in an infant with Coxsackie B2 viral infection  

SciTech Connect

After the intravenous administration of Tc-99m sulfur colloid, there was found homogeneous lung, renal, and splenic uptake with absence of uptake by the liver and bone marrow of a nine-day-old female infant. More than 20 other doses were dispensed from the same Tc-99m sulfur colloid preparation with the expected biodistribution. A necropsy done two days later showed diffuse hepatic hemorrhagic necrosis without evidence of intravascular fibrin deposition in the lungs or kidneys. The underlying cause of the infant's disease was a Coxsackie B2 viral infection, based upon positive postmortem viral cultures of kidney and liver tissues and characteristic histopathologic lesions of the central nervous system and viscera. The altered biodistribution presumably reflected marked impairment of Kupffer cell function and an apparent increase in pulmonary macrophages.

Hinkle, G.H.; Leonard, J.C.; Krous, H.F.; Alexander, J.B.



Cold Pressure Test Producing Coronary Spasm, Coronary Thrombosis and Myocardial Infarction in a Patient with IgM Antibodies against Coxsackie B Virus  

Microsoft Academic Search

Several lines of evidence have shown that viral infections are capable of causing coronary spasm and precipitating or mimicking clinical myocardial infarction. Here we report the case of a 41-year-old woman with recurrent angina who was admitted to our hospital because of ventricular tachycardia. Laboratory examination revealed positive IgM titers against Coxsackie B virus. Coronary angiography showed normal coronary arteries,

Werner Haberbosch; Norbert Roerich; Joerg Neuzner



Trace element distribution in heart tissue sections studied by nuclear microscopy is changed in coxsackie virus B3 myocarditis in methyl mercury-exposed mice  

Microsoft Academic Search

Methyl mercury (MeHg) has been shown to change Coxsackie virus type B3 (CB3) myocarditis in a direction compatible with the\\u000a development of chronic disease. Murine models of CB3 myocarditis closely mimic the pathogenesis in humans. There are also\\u000a indications that metals, such as mercury, and trace elements may interact and adversely affect viral replication and development\\u000a of inflammatory lesions. The

Nils-Gunnar Ilbäck; Ulf Lindh; Lars Wesslén; Jan Fohlman; Göran Friman



Identification of a putative shared epitope between Coxsackie virus B4 and alpha cardiac myosin heavy chain.  

PubMed Central

Molecular mimicry is an important postulated mechanism for autoimmunity in viral myocarditis. The 356-1 monoclonal antibody neutralizes Coxsackie virus B4 by binding to the VP1 protein and cross-reacts with mouse alpha cardiac myosin heavy chain. We used this monoclonal antibody to screen a lambda gt11 expression library made from CD-1 mouse hearts. Of the 48 positive plaques/10(6) recombinant phages examined, 14 of the strongest-reacting clones were purified for additional studies. The inserts were amplified by polymerase chain reaction and the amplified products ranged from about 150 to 1400 bp in size. Northern hybridization using these inserts demonstrated that 11 out of 14 reacted with a message equivalent to that of cardiac myosin in size. Additional Southern hybridization studies suggested that these 11 inserts contained overlapping sequences in the light meromyosin fragment of cardiac myosin. Sequence analysis confirmed that these 11 independent, recombinant clones contained a common sequence representing amino acid residues 1299-1647. Within this fragment only one isoform-specific site matched the observed reactivity pattern of 356-1 among hearts from various species. Thus, we were able to identify a putative shared epitope represented by residues 1632-1647. Images Fig. 1

Beisel, K W; Srinivasappa, J; Prabhakar, B S



Routine use of mu-antibody-capture ELISA for the serological diagnosis of Coxsackie B virus infections.  


The role of coxsackie B viruses (CBV) in myo/pericarditis has been well documented; however, interpretation of static high neutralising antibody titres in individual patients has always been difficult. In introducing the mu-antibody capture ELISA test for the detection of CBV-specific IgM, we hoped to overcome this problem. A regimen for the routine serological diagnosis of CBV infections was introduced, using the CBV IgM ELISA as a screening test, followed by neutralisation tests (NT) to confirm the positive results. Seven hundred and sixty patients and 304 healthy adult controls were tested. The percentage CBV IgM positive in each of the clinical categories myo/pericarditis (33%) chest pain (22%), myalgic encephalomyelitis (31%), myalgia/Bornholm (19%) and controls (9%) was similar to those found in previous studies using NT alone. Cross-reactions with other enteroviruses, including hepatitis A (Enterovirus 72), were observed but did not prove to be a problem in the illness studied, since most involved adults. Both homotypic and heterotypic CBV IgM responses were found. Matching IgM and NT indicated a recent CBV infection. Positive IgM with negative NT titres suggested a recent infection with an enterovirus other than a CBV. PMID:3016163

McCartney, R A; Banatvala, J E; Bell, E J



Transcription Factor Sp1 Is Involved in Expressional Regulation of Coxsackie and Adenovirus Receptor in Cancer Cells  

PubMed Central

Coxsackie and adenovirus receptor (CAR) was first known as a virus receptor. Recently, it is also known to have tumor suppressive activity such as inhibition of cell proliferation, migration, and invasion. It is important to understand how CAR expression can be regulated in cancers. Based on an existence of putative Sp1 binding site within CAR promoter, we investigated whether indeed Sp1 is involved in the regulation of CAR expression. We observed that deletion or mutation of Sp1 binding motif (?503/?498) prominently impaired the Sp1 binding affinity and activity of CAR promoter. Histone deacetylase inhibitor (TSA) treatment enhanced recruitment of Sp1 to the CAR promoter in ChIP assay. Meanwhile, Sp1 binding inhibitor suppressed the recruitment. Exogenous expression of wild-type Sp1 increased CAR expression in CAR-negative cells; meanwhile, dominant negative Sp1 decreased the CAR expression in CAR-positive cells. These results indicate that Sp1 is involved in regulation of CAR expression.

Chung, Sun-Ku; Kim, Joo-Young; Lim, Joong-Yeon; Park, Young Mi; Hwang, Ha-Young; Nam, Jae-Hwan; Park, Sang Ick



Genetic characterization of a Coxsackie A9 virus associated with aseptic meningitis in Alberta, Canada in 2010  

PubMed Central

Background An unusually high incidence of aseptic meningitis caused by enteroviruses was noted in Alberta, Canada between March and October 2010. Sequence based typing was performed on the enterovirus positive samples to gain a better understanding of the molecular characteristics of the Coxsackie A9 (CVA-9) strain responsible for most cases in this outbreak. Methods Molecular typing was performed by amplification and sequencing of the VP2 region. The genomic sequence of one of the 2010 outbreak isolates was compared to a CVA-9 isolate from 2003 and the prototype sequence to study genetic drift and recombination. Results Of the 4323 samples tested, 213 were positive for enteroviruses (4.93%). The majority of the positives were detected in CSF samples (n?=?157, 73.71%) and 81.94% of the sequenced isolates were typed as CVA-9. The sequenced CVA-9 positives were predominantly (94.16%) detected in patients ranging in age from 15 to 29 years and the peak months for detection were between March and October. Full genome sequence comparisons revealed that the CVA-9 viruses isolated in Alberta in 2003 and 2010 were highly homologous to the prototype CVA-9 in the structural VP1, VP2 and VP3 regions but divergent in the VP4, non-structural and non-coding regions. Conclusion The increase in cases of aseptic meningitis was associated with enterovirus CVA-9. Sequence divergence between the prototype strain of CVA-9 and the Alberta isolates suggests genetic drifting and/or recombination events, however the sequence was conserved in the antigenic regions determined by the VP1, VP2 and VP3 genes. These results suggest that the increase in CVA-9 cases likely did not result from the emergence of a radically different immune escape mutant.



No Evidence for an Association of Coxsackie Virus Infections during Pregnancy and Early Childhood with Development of Islet Autoantibodies in Offspring of Mothers or Fathers with Type 1 Diabetes  

Microsoft Academic Search

Recent case-control studies reported an increased frequency of antibodies against Coxsackie virus (CV) antigens in patients with newly diagnosed type 1 diabetes and during pregnancy in mothers of diabetic offspring, suggesting a role for CV infections in the pathogenesis of type 1 diabetes (T1D). However, it is not known whether CV infections are causally related to the development of islet

M Füchtenbusch; A Irnstetter; G Jäger; A.-G Ziegler



The transduction of Coxsackie and Adenovirus Receptor-negative cells and protection against neutralizing antibodies by HPMA-co-oligolysine copolymer-coated adenovirus  

PubMed Central

Adenoviral (AdV) gene vectors offer efficient nucleic acid transfer into both dividing and non-dividing cells. However issues such as vector immunogenicity, toxicity and restricted transduction to receptor-expressing cells have prevented broad clinical translation of these constructs. To address this issue, engineered AdV have been prepared by both genetic and chemical manipulation. In this work, a polymer-coated Ad5 formulation is optimized by evaluating a series of N-(2-hydroxypropyl) methacrylamide (HPMA)-co-oligolysine copolymers synthesized by living polymerization techniques. This synthesis approach was used to generate highly controlled and well-defined polymers with varying peptide length (K5, K10 and K15), polymer molecular weight, and degradability to coat the viral capsid. The optimal formulation was not affected by the presence of serum during transduction and significantly increased Ad5 transduction of several cell types that lack the Coxsackie and Adenovirus Receptor (CAR) by up to 6-fold compared to unmodified AdV. Polymer-coated Ad5 also retained high transduction capability in the presence of Ad5 neutralizing antibodies. The critical role of heparan sulfate proteoglycans (HSPGs) in mediating cell binding and internalization of polymer-coated AdV was also demonstrated by evaluating transduction in HSPG-defective recombinant CHO cells. The formulations developed here are attractive vectors for ex vivo gene transfer in applications such as cell therapy. In addition, this platform for adenoviral modification allows for facile introduction of alternative targeting ligands.

Wang, Chung-Huei K.; Chan, Leslie W.; Johnson, Russell N.; Chu, David S.H.; Shi, Julie; Schellinger, Joan G.; Lieber, Andre; Pun, Suzie H.



Coxsackie- and adenovirus receptor (CAR) is expressed in lymphatic vessels in human skin and affects lymphatic endothelial cell function in vitro  

SciTech Connect

Lymphatic vessels play an important role in tissue fluid homeostasis, intestinal fat absorption and immunosurveillance. Furthermore, they are involved in pathologic conditions, such as tumor cell metastasis and chronic inflammation. In comparison to blood vessels, the molecular phenotype of lymphatic vessels is less well characterized. Performing comparative gene expression analysis we have recently found that coxsackie- and adenovirus receptor (CAR) is significantly more highly expressed in cultured human, skin-derived lymphatic endothelial cells (LECs), as compared to blood vascular endothelial cells. Here, we have confirmed these results at the protein level, using Western blot and FACS analysis. Immunofluorescence performed on human skin confirmed that CAR is expressed at detectable levels in lymphatic vessels, but not in blood vessels. To address the functional significance of CAR expression, we modulated CAR expression levels in cultured LECs in vitro by siRNA- and vector-based transfection approaches. Functional assays performed with the transfected cells revealed that CAR is involved in distinct cellular processes in LECs, such as cell adhesion, migration, tube formation and the control of vascular permeability. In contrast, no effect of CAR on LEC proliferation was observed. Overall, our data suggest that CAR stabilizes LEC-LEC interactions in the skin and may contribute to lymphatic vessel integrity.

Vigl, Benjamin; Zgraggen, Claudia; Rehman, Nadia; Banziger-Tobler, Nadia E.; Detmar, Michael [Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology, ETH Zurich, Wolfgang-Pauli Str. 10, CH-8093 Zurich (Switzerland); Halin, Cornelia [Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology, ETH Zurich, Wolfgang-Pauli Str. 10, CH-8093 Zurich (Switzerland)], E-mail:



Distribution of antibodies against Coxsackie B viruses, arboviruses and Toxoplasma gondii among patients with endomyocardial fibrosis (EMF) compared with normal subjects from EMF endemic and non-endemic zones of Nigeria.  


The sera of eight endomyocardial fibrosis (EMF) subjects, 11 siblings of one of them and 16 normal children matched with the EMF patients for age, sex and socio-economic status from Ogunmakin and Shao/Oloru communities (eight each), situated in EMF-endemic and non-endemic areas of Nigeria respectively, were examined for the presence of antibodies against Coxsackie viruses B1-6, 16 arboviruses and Toxoplasma gondii. Sera from 36 other randomly selected normal children from Ogunmakin and 26 other randomly selected children from Shao/Oloru were also tested for the presence of antibodies against Toxoplasma gondii and the 16 arboviruses. None of the eight EMF subjects nor the 11 siblings of one of them had antibodies against any of the Coxsackie viruses B1-6 in their sera. Two of the 16 matched control subjects, one from each community, had positive antibodies, at equivocal titres against Coxsackie B1 (Ogunmakin) and B4 (Shao/Oloru). There was no significant difference in the distribution of antibody titres to the arboviruses between the EMF patients and matched controls. Normal children from the Shao/Oloru community had higher percentage antibody reactions and higher titres to the arboviruses compared with the children from Ogunmakin. All the eight EMF patients had high antibody titres against Toxoplasma gondii. Seven (87.5%) of the matched controls from Ogunmakin were sero-positive for Toxoplasma gondii compared with three (37.5%) of the matched controls from Shao/Oloru. Of the 36 normal children from Ogunmakin, 32 (88.9%) were sero-positive compared with 11 (42.3%) of the 26 normal children from Shao/Oloru. Four (36.4%) of the 11 siblings of one of the EMF patients had weak sero-positivity. It is therefore concluded that further studies are needed to clarify the role, if any, of Toxoplasma gondii in EMF. PMID:2165348

Ijaola, O; Falase, A O



[Anti-Coxsackie antibodies and congestive cardiomyopathies].  


During a 4-years period 122 patients admitted to the cardiologic hospital in Lille presented a significative positivity of serological reaction against Coxsackievirus B. Among these patients 19 suffered from acute pericarditis, 4 from cardiac arrhythmias, 3 from Bornholm syndrome and 14 from apparently primary congestive cardiomyopathy. In most of these 14 patients representing 18.4% of congestive cardiomyopathies investigated at the same time, hemodynamic studies and heart scanning with radioactive gallium were performed. 7 of these patients had a chronic (group A) and 7 an acute evolution (group B). In group A the scans were negative 4 times out of 5, whereas in group B they were positive in 5 out of 5 examinations. The control group including 3247 patients coming from the Institut Pasteur exhibited serological positivity in 4.6% of cases. Comparison of this group with that of cardiomyopathies shows a significant difference (p less than 0.001). The authors draw the attention to the frequency of positive serodiagnosis in congestive cardiomyopathy and stress the importance of gallium scintiscanning. PMID:3099680

Leroy, O; Asseman, P; Traisnel, G; Durand, P; Beuscart, R; Dewilde, A; Wattré, P; Théry, C



Comparative genomics of the coxsackie B viruses and related enteroviruses.  


Genomic analysis of the group B coxsackieviruses (CVB) has improved our understanding of CVB evolution, epidemiology, and pathogenesis. Comparison of capsid sequence alignments and virion structures allows correlation of capsid diversity with surface features, such as loops, the receptor canyon, and antigenic sites. Pairwise sequence comparisons and phylogenetic analyses can be used to rapidly identify and classify enteroviruses. Enteroviruses are monophyletic by type only within the capsid region. The CVBs as a group are monophyletic in the capsid region, probably due to their shared use of the coxsackievirus-adenovirus receptor (other members of HEV-B use different receptors). Outside the capsid region, enteroviruses are monophyletic only by species (not by type), reflecting a high frequency of intertypic recombination within a species. Further genomic studies, accompanied by well-characterized clinical outcome/disease data, will facilitate fine-scale mapping of genetic determinants that contribute to virulence. PMID:18357764

Oberste, M S



Coxsackie virus infection of the placenta associated with neurodevelopmental delays in the newborn  

Microsoft Academic Search

OBJECTIVE:To determine if viral infection of the placenta was associated with long-term neurodevelopmental delays in the newborn.METHODS:Placental tissue from seven newborn infants with severe respiratory failure and subsequent neurodevelopmental abnormalities as well as ten normal controls and five cases of known placental infection (cytomegalovirus, herpes simplex virus, and parvovirus) were tested by in situ hybridization or reverse transcriptase in situ

Elizabeth Euscher; Jonathan Davis; Ian Holzman; Gerard J Nuovo



Synergistic drug combinations against the in vitro replication of Coxsackie B1 virus  

Microsoft Academic Search

The existence of synergistic drug combinations against the in vitro replication of poliovirus type 1 (Mahoney) (PV-1) had been established in our previous work. The objective of the present study was to test the combined effects of the different drugs against another representative of the enterovirus genus, i.e. Coxsackievirus B1 (CBV-1). Dual combinations of enviroxime, disoxaril, arildone, PTU-23, HBB and

Lubomira Nikolaeva-Glomb; Angel S Galabov



The Murine CAR Homolog Is a Receptor for Coxsackie B Viruses and Adenoviruses  

Microsoft Academic Search

Complementary DNA clones encoding the murine homolog (mCAR) of the human coxsackievirus and adenovirus receptor (CAR) were isolated. Nonpermissive CHO cells transfected with mCAR cDNA became susceptible to infection by coxsackieviruses B3 and B4 and showed increased susceptibility to adenovirus- mediated gene transfer. These results indicate that the same receptor is responsible for virus interactions with both murine and human




Distinct different expression of Th17 and Th9 cells in coxsackie virus B3-induced mice viral myocarditis  

Microsoft Academic Search

Background  Recently, a new subset of CD4+T helper(Th) cell that predominantly secret cytokine interleukin-9(IL-9) is identified, termed Th9 cell. It has been reported\\u000a to participate in tissue inflammation and autoimmune responses, and induce disease which differed from Th17 cells. Th17 cells\\u000a have been shown to play a critical role in viral myocarditis (VMC), but whether Th9 cells are involved in the

Kong Qing; Wu Weifeng; Yang Fan; Yan Yuluan; Pang Yu; Huang Yanlan



Efficient Infection Mediated by Viral Receptors Incorporated into Retroviral Particles  

PubMed Central

Many host cell surface proteins, including viral receptors, are incorporated into enveloped viruses. To address the functional significance of these host proteins, murine leukemia viruses containing the cellular receptors for Rous sarcoma virus (Tva) or ecotropic murine leukemia virus (MCAT-1) were produced. These receptor-pseudotyped viruses efficiently infect cells expressing the cognate viral envelope glycoproteins, with titers of up to 105 infectious units per milliliter for the Tva pseudotypes. Receptor and viral glycoprotein specificity and functional requirements are maintained, suggesting that receptor pseudotype infection recapitulates events of normal viral entry. The ability of the Tva and MCAT-1 pseudotypes to infect cells efficiently suggests that, in contrast to human immunodeficiency virus type 1 entry, neither of these retroviral receptors requires a coreceptor for membrane fusion. In addition, the ability of receptor pseudotypes to target infected cells suggests that they may be useful therapeutic reagents for directing infection of viral vectors. Receptor-pseudotyped viruses may be useful for identifying new viral receptors or for defining functional requirements of known receptors. Moreover, this work suggests that the production of receptor pseudotypes in vivo could provide a mechanism for expanded viral tropism with potential effects on the pathogenesis and evolution of the virus.

Balliet, John W.; Bates, Paul



Histone deacetylase inhibitors restore cell surface expression of the coxsackie adenovirus receptor and enhance CMV promoter activity in castration-resistant prostate cancer cells.  


Adenoviral gene therapy using the death receptor ligand TRAIL as the therapeutic transgene can be safely administered via intraprostatic injection but has not been evaluated for efficacy in patients. Here we investigated the efficacy of adenoviral TRAIL gene therapy in a model of castration resistant prostate cancer and found that intratumoral injections can significantly delay tumor growth but cannot eliminate established lesions. We hypothesized that an underlying cause is inefficient adenoviral delivery. Using the LNCaP progression model of prostate cancer we show that surface CAR expression decreases with increasing tumorigenicity and that castration resistant C4-2b cells were more difficult to transduce with adenovirus than castration sensitive LNCaP cells. Many genes, including CAR, are epigenetically silenced during transformation but a new class of chemotherapeutic agents, known as histone deacetylase inhibitors (HDACi), can reverse this process. We demonstrate that HDACi restore CAR expression and infectivity in C4-2b cells and enhance caspase activation in response to infection with a TRAIL adenovirus. We also show that in cells with high surface CAR expression, HDACi further enhance transgene expression from the CMV promoter. Thus HDACi have multiple beneficial effects, which may enhance not only viral but also non-viral gene therapy of castration resistant prostate cancer. PMID:22288017

Kasman, Laura; Onicescu, Georgiana; Voelkel-Johnson, Christina



Histone Deacetylase Inhibitors Restore Cell Surface Expression of the Coxsackie Adenovirus Receptor and Enhance CMV Promoter Activity in Castration-Resistant Prostate Cancer Cells  

PubMed Central

Adenoviral gene therapy using the death receptor ligand TRAIL as the therapeutic transgene can be safely administered via intraprostatic injection but has not been evaluated for efficacy in patients. Here we investigated the efficacy of adenoviral TRAIL gene therapy in a model of castration resistant prostate cancer and found that intratumoral injections can significantly delay tumor growth but cannot eliminate established lesions. We hypothesized that an underlying cause is inefficient adenoviral delivery. Using the LNCaP progression model of prostate cancer we show that surface CAR expression decreases with increasing tumorigenicity and that castration resistant C4-2b cells were more difficult to transduce with adenovirus than castration sensitive LNCaP cells. Many genes, including CAR, are epigenetically silenced during transformation but a new class of chemotherapeutic agents, known as histone deacetylase inhibitors (HDACi), can reverse this process. We demonstrate that HDACi restore CAR expression and infectivity in C4-2b cells and enhance caspase activation in response to infection with a TRAIL adenovirus. We also show that in cells with high surface CAR expression, HDACi further enhance transgene expression from the CMV promoter. Thus HDACi have multiple beneficial effects, which may enhance not only viral but also non-viral gene therapy of castration resistant prostate cancer.

Kasman, Laura; Onicescu, Georgiana; Voelkel-Johnson, Christina



Trans-complementation of vector replication versus Coxsackie-adenovirus-receptor overexpression to improve transgene expression in poorly permissive cancer cells.  


Gene therapy of cancer requires high-level expression of therapeutic transgenes in the target cells. Poor gene transfer is an important limitation to adenovector-mediated cancer gene therapy. We investigated two fundamentally different approaches to improve transgene expression in poorly permissive cancer cells. First, overexpression of the adenovirus attachment receptor CAR to facilitate receptor-mediated adenovector (AdV) uptake into the target cells; second, co-infection of this vector together with traces of replication competent adenovirus (RCA) accidentally arising by back-recombination during large-scale vector preparation. Among eight gastrointestinal cancer cell lines, the colorectal cancer lines showed particularly poor vector-mediated transgene expression (down to 67-fold lower than in HeLa cells). Expression of the adenovirus receptors CAR, alpha(v)beta5- and alpha(v)beta3-integrin were highly variable between cell lines. AdV uptake was significantly associated with CAR levels on the cell surface, but not with those of the integrins. AdV-mediated CAR overexpression increased CAR density on the surface of all investigated tumor cells and led to enhancement of transgene expression by 1.8- to 6.7-fold. The other principle to enhance transgene expression was 'trans-complementation' of the therapeutic vector, ie induction of its replication within the target cells. Traces of RCA in a vector preparation, as well as purified RCA were found to provide sufficient E1-region transcripts to induce replication of the therapeutic vector genome. The number of adenovector-based transgene expression cassettes was greatly amplified by this principle, notably without any influence on the rate of vector entry. Co-infection of four colorectal cancer cell lines with marker vector plus RCA (at around 240:1 particle ratio) resulted in far stronger enhancement of transgene expression (up to 46-fold) as compared with CAR overexpression, even in cancers almost refractory to standard adenovector-mediated gene transfer. Whereas RCAs need to be strictly avoided in gene therapy of non-malignant diseases for safety reasons, the magnitude of helper virus-induced therapeutic transgene expression could possibly warrant application of this principle to overcome the resistance of highly malignant cancers against gene therapy. PMID:11127584

Fechner, H; Wang, X; Wang, H; Jansen, A; Pauschinger, M; Scherübl, H; Bergelson, J M; Schultheiss, H P; Poller, W



[Oxidative stress in patients with type I diabetes mellitus and persistent coxsackie virus B infection as the reason of dysfunction of the immune system].  


A chronic inflammatory process takes place in patients with diabetes mellitus type 1. Numerous disorders of the immune status and complications testify the present of this process. The presence of chronic inflammation at diabetes mellitus enhances free radical reactions which are accompanied by oxidative stress. PMID:20455442

Hyrin, V V


The Coxsackie-Adenovirus Receptor (CAR) Is Used by Reference Strains and Clinical Isolates Representing All Six Serotypes of Coxsackievirus Group B and by Swine Vesicular Disease Virus  

Microsoft Academic Search

Group B coxsackieviruses are etiologically linked to many human diseases, and cell surface receptors are postulated to play an important role in mediating their pathogenesis. The coxsackievirus adenovirus receptor (CAR) has been shown to function as a receptor for selected strains of coxsackievirus group B (CVB) serotypes 3, 4, and 5 and is postulated to serve as a receptor for

Tami A. Martino; Martin Petric; Hana Weingartl; Jeffrey M. Bergelson; Mary Anne Opavsky; Christopher D. Richardson; John F. Modlin; Robert W. Finberg; Kevin C. Kain; Norman Willis; Charles J. Gauntt; Peter P. Liu



Coxsackie B virus infection and ? cell autoantibodies in newly diagnosed IDDM adult patients 1 The molecular results presented in this article have been partially published in J Med Virol 1997;52:121–127. 1  

Microsoft Academic Search

Background: Environmental agents such as viruses have been identified as potentially important determinants of insulin-dependent diabetes mellitus (IDDM). Enterovirus infections, Coxsackievirus B especially, could be linked to the ? cell damaging process and to the onset of clinical IDDM.Objectives: Enteroviral (EV) infection and ? cell autoimmunity were studied in adult patients at the onset of IDDM.Study design: A total of

Laurent Andréoletti; Didier Hober; Christine Hober-Vandenberghe; Isabelle Fajardy; Sandrine Belaich; Valérie Lambert; Marie-Christine Vantyghem; Jean Lefebvre; Pierre Wattre



Genetic diversity of enterovirus subgroups  

Microsoft Academic Search

Summary Enterovirus serotypes were studied using nucleic acid hybridization and nucleotide sequence analysis. A great majority of enteroviruses could be roughly divided into two larger subgroups the first consisting of poliovirus and certain coxsackievirus A serotypes. The second subgroup included coxsackie B viruses, most ECHO viruses, enterovirus 71 and representatives of coxsackie A viruses. Enterovirus 70 showed low homology to

P. Auvinen; G. Stanway; T. Hyypiä



Targeting of Adenovirus Vectors to Breast Cancer Mediated by Soluble Receptor-Ligand Fusion Proteins.  

National Technical Information Service (NTIS)

The use of adenovirus (Ad) vectors for cancer gene therapy is currently limited by several factors, including broad Ad tropism associated with expression of coxsackie virus and adenovirus receptor (CAR) in normal human tissues, and limited CAR levels in t...

I. P. Dmitriev E. A. Kashentseva



[Outbreak of acute enterovirus intestinal infection in Sakhalin region in August 2010].  


The investigation of cases of acute intestinal infections in the Sakhalin region of Russia in August, 2010 is described. Epidemiological and molecular biological studies were conducted. After initial PCR screening and determining the nucleotide sequences of the positive samples the following enteroviruses were found: Coxsackie A2 - 42 samples (45%), Coxsackie A4--31 sample (34%), Enterovirus 71--6 samples (6,5%), Coxsackievirus B5--6 samples (6,5%), Coxsackie B3--4 samples (4%) and Coxsackie B1--4 samples (4%). The phylogenetic analysis of sequences showed that the closest analogues for the nucleotide sequences of these genotypes were previously identified in Japan, Korea and China in 2000-2010. PMID:22642180

Demina, A V; Ternovo?, V A; Darizhapov, B B; Iakubich, T V; Sementsova, A O; Demina, O K; Protopopova, E V; Loktev, V B; Agafonov, A P; Netesov, S V



Identification and Detection of Water-Borne Viruses by Immunoenzymatic Methods.  

National Technical Information Service (NTIS)

A quantitative enzyme-linked immunosorbent assay (ELISA) was used for identification of viruses selected as representative water-borne viruses: poliovirus 1, echovirus 6, coxsackievirus A9, and coxsackie B viruses. Partially purified viral antigens or vir...

J. E. Herrmann



Production of Enterovirus Antisera.  

National Technical Information Service (NTIS)

The production of enterovirus antisera in horses and of rhinovirus antisera in goats is reported. Eight other coxsackie virus, one echo virus, and eight rhinovirus antigens are in progress. Data on inoculations, bleedings, and stored and transferred sera ...

R. W. Brown




Microsoft Academic Search

Enteroviruses (EVs) comprise a genus in the Picornaviridae family, so named because they are small, single-stranded ribonucleic\\u000a acid (RNA)-containing viruses. The EVs are divided into the subgroups of polioviruses, coxsackie A viruses, coxsackie B viruses,\\u000a and echoviruses based on replication properties in tissue culture and animal models. Newer EVs are simply designated by number\\u000a (e.g., EV 71), rather than by

Mark J. Abzug


Altering the Ad5 Packaging Domain Affects the Maturation of the Ad Particles  

PubMed Central

We have previously described a new family of mutant adenoviruses carrying different combinations of attB/attP sequences from bacteriophage PhiC31 flanking the Ad5 packaging domain. These novel helper viruses have a significantly delayed viral life cycle and a severe packaging impairment, regardless of the presence of PhiC31 recombinase. Their infectious viral titers are significantly lower (100–1000 fold) than those of control adenovirus at 36 hours post-infection, but allow for efficient packaging of helper-dependent adenovirus. In the present work, we have analyzed which steps of the adenovirus life cycle are altered in attB-helper adenoviruses and investigated whether these viruses can provide the necessary viral proteins in trans. The entry of attB-adenoviral genomes into the cell nucleus early at early timepoints post-infection was not impaired and viral protein expression levels were found to be similar to those of control adenovirus. However, electron microscopy and capsid protein composition analyses revealed that attB-adenoviruses remain at an intermediate state of maturation 36 hours post-infection in comparison to control adenovirus which were fully mature and infective at this time point. Therefore, an additional 20–24 hours were found to be required for the appearance of mature attB-adenovirus. Interestingly, attB-adenovirus assembly and infectivity was restored by inserting a second packaging signal close to the right-end ITR, thus discarding the possibility that the attB-adenovirus genome was retained in a nuclear compartment deleterious for virus assembly. The present study may have substantive implications for helper-dependent adenovirus technology since helper attB-adenovirus allows for preferential packaging of helper-dependent adenovirus genomes.

Alba, Raul; Cots, Dan; Ostapchuk, Philomena; Bosch, Assumpcio; Hearing, Patrick; Chillon, Miguel



144. Magnetically Responsive Nanoparticles Enhance Adenoviral Gene Delivery in Cultured Smooth Muscle Cells  

Microsoft Academic Search

Replication deficient adenoviruses (Ad) are extensively investigated for gene delivery applications due to a number of advantages as gene vectors, including high transgene expression and their ability to transduce both quiescent and dividing cells. Their clinical use is however limited by therapeutically suboptimal transduction levels in cell types expressing low levels of Coxsackie-Ad receptor (CAR), as well as systemic toxicity

Michael Chorny; Ilia Fishbein; Ivan Alferiev; Origene Nyanguile; Robert J. Levy



Enhanced delivery of mda-7\\/IL24 using a serotype chimeric adenovirus (Ad.5\\/3) improves therapeutic efficacy in low CAR prostate cancer cells  

Microsoft Academic Search

Gene therapy is being examined as a potential strategy for treating prostate cancer. Serotype 5 adenovirus (Ad.5) is routinely used as a vector for transgene delivery. However, the infectivity of Ad.5 is dependent on Coxsackie-adenovirus receptors (CARs); many tumor types show a reduction in this receptor in vivo, thereby limiting therapeutic gene transduction. Serotype chimerism is one approach to circumvent

R Dash; I Dmitriev; Z-z Su; S K Bhutia; B Azab; N Vozhilla; A Yacoub; P Dent; D T Curiel; D Sarkar; P B Fisher



Replication or Inactivation of Different Viruses by Human Lymphocyte Preparations  

PubMed Central

Cultures of phytohemagglutinin-stimulated, unstimulated, and frozen and thawed mononuclear (as controls) cells from normal human donors were infected with different viruses. A variable pattern of virus infectivity was noted, and influenza and Coxsackie virus were rapidly inactivated even by stimulated lymphocytes. Direct inactivation by lymphocytes may be one form of host defense in infection by some viruses.

Denman, A. M.; Rager-Zisman, B.; Merigan, T. C.; Tyrrell, D. A. J.



Investigation to Increase the Viricidal Capacity of Disinfectant, Germicidal and Fungicidal, Phenolic, Dry-Type.  

National Technical Information Service (NTIS)

The straight O-D-1435 Type II disinfectant had little capacity to inactivate the poliovirus type I when used in concentrations up to 15,000 mg/l with contact times up to 24 hours. The Coxsackie A9 and Echo 12 viruses showed no inactivation with 10,000 mg/...

O. J. Sproul



Realms of the Viruses Online  

ERIC Educational Resources Information Center

|Viruses have evolved strategies for infecting all taxa, but most viruses are highly specific about their cellular host. In humans, viruses cause diverse diseases, from chronic but benign warts, to acute and deadly hemorrhagic fever. Viruses have entertaining names like Zucchini Yellow Mosaic, Semliki Forest, Coxsackie, and the original…

Liu, Dennis



Some Criteria for Judging the Relationship Between Bacteria and Viruses. I. E. Coli Bacteria as Adsorbents of Enteroviruses.  

National Technical Information Service (NTIS)

Different E. coli strains are capable of adsorbing Coxsackie B3, B5, and poliomyelitis viruses. Virus adsorbed on bacteria can be detected from the cytopathic effect in tissue culture and by the plaque assay procedure (the amount of virus will be expresse...

A. E. Essel K. G. Medvinskaya I. I. Neznanskaya



Intercellular Trafficking of Adenovirus-Delivered HSV VP22 from the Retinal Pigment Epithelium to the Photoreceptors—Implications for Gene Therapy  

Microsoft Academic Search

Adenovirus (Ad)-mediated gene transfer is a promising technology for therapy of a wide variety of genetic disorders of the retina. The tropism of Ad vectors limits their utility to cells that express the coxsackie–adenovirus receptor. Upon ocular delivery, Ad vectors primarily infect the retinal pigment epithelium (RPE) and the Müller cells of the retina. However, the most frequent blinding diseases

Siobhan M. Cashman; Sonia L. Sadowski; David J. Morris; Jeanne Frederick; Rajendra Kumar-Singh




EPA Science Inventory

The inactivation kinetics of ClO2 on two enteroviruses, poliovirus 1 (Mahoney) and coxsackie virus A9, and an enteric indicator of fecal pollution, Escherichia coli, were examined in laboratory studies. In addition, the disinfecting ability of ClO2 as affected by particulates (bo...


879. Transduction Enhancement by the Adenovirus\\/Polymer Complexes Derivatized with Protein Transduction Domain Peptides  

Microsoft Academic Search

Background: Cellular uptake of group C adenoviruses critically depends on the availability of the cognate Coxsackie Adenovirus Receptors (CARs) on the cell surface. In cardiovascular tissues paucity of CARs is a limiting factor for the successful application of adenoviral vectors. Additionally, immune responses elicited by Ad infection diminish transgene expression due to the elimination of free Ad and transduced cells.

Ilia Fishbein; Ivan S. Alferiev; Richard Gaster; Michael Chorny; Origene Nyanguile; Robert J. Levy



415. Evaluating Polyethylene Glycol (PEG) Modified Adenoviral Vectors for Cell Targeting and Detargeting and Reducing Vector Toxicity  

Microsoft Academic Search

Adenovirus type 5 (Ad5) are robust gene delivery vectors in vivo, but are limited by the number of clinically relevant tissues they can transduce effectively, mostly due to the low expression of coxsackie and adenoviral receptor (CAR) in most tissues. In order to improve gene transfer to tissues that express low level of CAR, vectors are detargeted to remove the

Hoyin Mok; Michael A. Barry



Infections and autoimmune diseases  

Microsoft Academic Search

The high percentage of disease-discordant pairs of monozygotic twins demonstrates the central role of environmental factors in the etiology of autoimmune diseases. Efforts were first focussed on the search for triggering factors. The study of animal models has clearly shown that infections may trigger autoimmune diseases, as in the case of Coxsackie B4 virus in type I diabetes and the

Jean-François Bach



CAR regulates epithelial cell junction stability through control of E-cadherin trafficking.  


CAR (Coxsackie and Adenovirus Receptor) is the primary docking receptor for typeB coxsackie viruses and subgroup C adenoviruses. CAR is a member of the JAM family of adhesion receptors and is located to both tight and adherens junctions between epithelial cells where it can assemble adhesive contacts through homodimerisation in trans. However, the role of CAR in controlling epithelial junction dynamics remains poorly understood. Here we demonstrate that levels of CAR in human epithelial cells play a key role in determining epithelial cell adhesion through control of E-cadherin stability at cell-cell junctions. Mechanistically, we show that CAR is phosphorylated within the C-terminus by PKC? and that this in turn controls Src-dependent endocytosis of E-cadherin at cell junctions. This data demonstrates a novel role for CAR in regulating epithelial homeostasis. PMID:24096322

Morton, Penny E; Hicks, Alexander; Nastos, Theodoros; Santis, George; Parsons, Maddy



Serum immunoreactive trypsin concentrations in infectious and non-infectious illnesses and in juvenile diabetes.  


Serum immunoreactive trypsin (SIT) concentrations were measured in 244 patients with infectious illnesses and in 281 children with diabetes of recent onset. Results were compared with reference ranges established in 107 patients with non-infectious, non-diabetic illnesses, in whom SIT concentrations were found to increase with advancing age. Reduced or undetectable concentrations of SIT were associated with diabetes in children and with a few cases of severe childhood infection. Increased SIT concentrations were associated with virologically confirmed cases of infection with mumps and Coxsackie B virus infection, and with clinical diagnoses of mumps, PUO, and meningitis in children, and with Bornholm disease, cardiac infection, and respiratory infection in adults. It is suggested that silent invasion of the exocrine pancreas with elevation of the SIT concentration may accompany infection by Coxsackie B, mumps, and, possibly, other viruses. PMID:512051

Gamble, D R; Moffatt, A; Marks, V



CAR regulates epithelial cell junction stability through control of E-cadherin trafficking  

PubMed Central

CAR (Coxsackie and Adenovirus Receptor) is the primary docking receptor for typeB coxsackie viruses and subgroup C adenoviruses. CAR is a member of the JAM family of adhesion receptors and is located to both tight and adherens junctions between epithelial cells where it can assemble adhesive contacts through homodimerisation in trans. However, the role of CAR in controlling epithelial junction dynamics remains poorly understood. Here we demonstrate that levels of CAR in human epithelial cells play a key role in determining epithelial cell adhesion through control of E-cadherin stability at cell-cell junctions. Mechanistically, we show that CAR is phosphorylated within the C-terminus by PKC? and that this in turn controls Src-dependent endocytosis of E-cadherin at cell junctions. This data demonstrates a novel role for CAR in regulating epithelial homeostasis.

Morton, Penny E.; Hicks, Alexander; Nastos, Theodoros; Santis, George; Parsons, Maddy



Preclinical Characterization of the Antiglioma Activity of a Tropism-Enhanced Adenovirus Targeted to the Retinoblastoma Pathway  

Microsoft Academic Search

Background: Oncolytic adenoviruses are promising thera- pies for the treatment of gliomas. However, untargeted viral replication and the paucity of coxsackie-adenovirus recep- tors (CARs) on tumor cells are major stumbling blocks for adenovirus-based treatment. We studied the antiglioma ac- tivity of the tumor-selective Delta-24 adenovirus, which en- compasses an early 1 A adenoviral (E1A) deletion in the retinoblastoma (Rb) protein-binding

Juan Fueyo; Ramon Alemany; Candelaria Gomez-Manzano; Gregory N. Fuller; Asadullah Khan; Charles A. Conrad; Ta-Jen Liu; Hong Jiang; Michael G. Lemoine; Kaori Suzuki; Raymond Sawaya; David T. Curiel; W. K. Alfred Yung; Frederick F. Lang



Treatment of Ovarian Cancer with a Tropism Modified Oncolytic Adenovirus1  

Microsoft Academic Search

Ad5-24RGD is an adenovirus that is selectively replication competent in cells defective in the Rb\\/p16 pathway, such as ovarian cancer cells. The fiber of Ad5-24RGD contains an integrin binding RGD-4C motif, allow- ing Coxsackie adenovirus receptor-independent infection of cancer cells. Oncolysis of cell lines was similar to that of a wild-type control, and replication in primary tumor material was shown

Gerd J. Bauerschmitz; John T. Lam; Anna Kanerva; Kaori Suzuki; Dirk M. Nettelbeck; Igor Dmitriev; Victor Krasnykh; Galina V. Mikheeva; Mack N. Barnes; Ronald D. Alvarez; Peter Dall; Ramon Alemany; David T. Curiel; Akseli Hemminki



Infectivity-Enhanced Adenoviruses Deliver Efficacy in Clinical Samples and Orthotopic Models of Disseminated Gastric Cancer  

Microsoft Academic Search

Purpose: Metastatic gastric cancer remains a common and devastating disease without curative treatment. Recent proof-of-concept clinical trials have validated gene therapy with adenoviruses as an effective and safe modality for the treatment of cancer. However, expression of the primary coxsackie-adenovirus receptor is variable in advanced cancers, and therefore, the use of hetero- logous receptors could be advantageous. Experimental Design: Here,

Lotta Kangasniemi; Tuula Kiviluoto; Anna Kanerva; Mari Raki; Tuuli Ranki; Merja Sarkioja; Hongju Wu; Frank Marini; Krister Ho; Helena Isoniemi; Henrik Alfthan; David T. Curiel; Akseli Hemminki



Post-Streptococcal Autoimmune Sequelae: A Link Between Infection and Autoimmunity  

Microsoft Academic Search

\\u000a For many years microbial pathogens have been implicated in the development of autoimmunity [1–8]. Several studies have attempted to link viral infections with rheumatoid arthritis (RA), insulin-dependent diabetes (I DDM),\\u000a myasthenia gravis, systemic lupus erythematosus (SLE), myocarditis and AIDS [2,8–15]. Candidate viruses included Coxsackie virus, Epstein-Barr virus, Herpes simplex virus, parvovirus, Type C retroviruses,\\u000a vesicular stomatitis virus and rubella virus

Malak Kotb


Viruses as a triggering factor of type 1 diabetes and genetic markers related to the susceptibility to the virus-associated diabetes.  


Type 1 diabetes results from the destruction of insulin-producing pancreatic beta cells. Genetic and environmental factors are implicated in the beta cell destruction. As environmental factors affecting the induction of type 1 diabetes, diabetogenic viruses, chemicals, toxins, and diet are likely candidates as either primary injurious agents of beta cells or triggering agents for the induction of autoimmunity. Regarding viruses as a triggering factor of type 1 diabetes, there are at least two different pathogenic mechanisms in virus-induced diabetes: cytolytic infection of beta cells, leading to their destruction, and triggering of autoimmunity, leading to the autoimmune-mediated destruction of beta cells. Since there is no correlation between the induction of antibodies to Coxsackie B viruses and the presence of islet cell autoantibodies in patients with type 1 diabetes, the induction of diabetes by Coxsackie B viruses may be due to cytolytic infection of beta cells rather than an autoimmune response. In contrast, rubella virus and cytomegalovirus (CMV) do appear to be somehow associated with autoimmune type 1 diabetes since there is a strong correlation between the presence of islet cell autoantibodies and persistent infections. Regarding genetic factors, there are distinct markers related to the susceptibility to Coxsackie B4 virus-associated type 1 diabetes and CMV-associated type 1 diabetes. Four specific DNA restriction endonuclease fragments (BamHI-DQ-beta 6.6, TaqI-DR-beta 4.3, TaqI-DR-beta 2.5 and TaqI-DR-beta 1.5 kb) are related to the susceptibility to Coxsackie B4 virus-associated type 1 diabetes while six specific DNA restriction endonuclease fragments (BamHI-DQ-alpha 12.5, -beta 3.7 and -beta 3.2 kb, TaqI-DQ-alpha 7.2, -beta 7.2 and -beta 5.4 kb) are related to the susceptibility to CMV-associated type 1 diabetes. PMID:2680367

Yoon, J W; Ihm, S H; Kim, K W



[The use of the natural mineral saponite for water decontamination].  


Sorption properties were studied of natural and activated specimens of saponite with respect to poliomyelitis virus, Coxsackie B 1 and B 6 viruses as well as to Enterobacteriaceae group bacteria. With the purpose of comparing the processes of sorption of microorganisms, other minerals were also used, such as bentonite, alunite, glauconite, ceolite. The natural saponite adsorptive properties were found out to undergo changes during the process of thermoactivation. Mechanisms are discussed of a decontaminating effect of thermoactivated saponite in water. PMID:10423991

Hyrin, V M; Bo?ko, I I; Rudychenko, V F


[A new sorbent for concentrating viruses from an aqueous medium].  


Sorption activity was studied of a naturally occurring dispersive mineral saponite towards certain representatives of the families of Picornaviridae and Reoviridae: poliomyelitis type II viruses, Coxsackie B 1 and B 6, monkey disease rotavirus. In increasing rank order in respect of specific sorption activity is naturally occurring bentonite--sodium form of bentonite--naturally occurring saponite. Possible mechanisms of sorption of viruses on the sorbent's surface, related to structural and chemical properties of the mineral are discussed. PMID:8630798

Hyrin, V M; Bo?ko, I I; Pluhatyr, V M; Rudychenko, V F; Derev'ianko, V M; Berezovs'ky?, S O; Trypil's'ka, L S; Bo?ko, O I


Synthesis of 5-isoxazol-5-yl-2?-deoxyuridines exhibiting antiviral activity against HSV and several RNA viruses  

Microsoft Academic Search

This paper describes a simple method for synthesizing a small library of 5-isoxazol-5-yl-2?-deoxyuridines from 5-iodo-2?-deoxyuridine. Nitrile oxides were generated in situ from oximes using a commercial bleaching agent; their cycloaddition with 5-ethynyl-2?-deoxyuridine yielded isoxazoles possessing activity against herpes simplex viruses 1 and 2, Encephalomyocarditis virus, Coxsackie B3, and vesicular stomatitis virus; these isoxazoles were, however, inactive against corona virus, influenza

Yoon-Suk Lee; Sun Min Park; Byeang Hyean Kim



Synthesis, antiviral activity and cytotoxicity evaluation of Schiff bases of some 2-phenyl quinazoline-4(3)H-ones  

Microsoft Academic Search

A new series of 3-(benzylideneamino)-2-phenylquinazoline-4(3H)-ones were prepared through Schiff base formation of 3-amino-2-phenyl quinazoline-4(3)H-one with various substituted carbonyl compounds. Their chemical structures were elucidated by spectral studies. Cytotoxicity and antiviral activity were evaluated against herpes simplex virus-1 (KOS), herpes simplex virus-2 (G), vaccinia virus, vesicular stomatitis virus, herpes simplex virus-1 TK- KOS ACVr, para influenza-3 virus, reovirus-1, Sindbis virus, Coxsackie

Krishnan Suresh Kumar; Swastika Ganguly; Ravichandran Veerasamy; Erik De Clercq



Construction of Urothelium-Specific Recombinant Adenovirus and Its Inhibition in Bladder Cancer Cell  

Microsoft Academic Search

Aim: To construct urothelium-specific recombinant adenovirus and investigate its inhibition in bladder cancer cell. Methods: RT-PCR analysis was used to determine expression patterns of hUPII and coxsackie adenovirus receptor on multiple cell lines. Transient transfection and luciferase detecting assay were used to detect tissue specificity of the hUPII promoter. Recombinant adenovirus Ad-UPII-E1A and Ad-UPII-Null were constructed. Restrictive enzyme digestion assay

Xiang-Dong He; Zhi-Ping Wang; Hai-Yan Wei; Qin Zhou; De-Gui Wang; Jun-Qiang Tian; Sheng-Jun Fu; Ronald Rodriguez



Persistence of enteroviral RNA in chronic fatigue syndrome is associated with the abnormal production of equal amounts of positive and negative strands of enteroviral RNA  

Microsoft Academic Search

A subgenomic restriction fragment from cDNA pre- pared from Coxsackie B2 virus (CVB2) RNA was snbcloned into a riboprobe vector allowing the produc- tion of enteroviral group-specific RNA probes comple- mentary to either the positive (genomic) or negative (template) strand of enteroviral RNA. These ribo- probes were used to follow productive infection of cultured cells by CVB2; as expected, positive

Louise Cunningham; N. E. Bowles; V. Dubowitz; L. C. Archard



Interaction between Mouse Adenovirus Type 1 and Cell Surface Heparan Sulfate Proteoglycans  

Microsoft Academic Search

Application of human adenovirus type 5 (Ad5) derived vectors for cancer gene therapy has been limited by the poor cell surface expression, on some tumor cell types, of the primary Ad5 receptor, the coxsackie-adenovirus-receptor (CAR), as well as the accumulation of Ad5 in the liver following interaction with blood coagulation factor X (FX) and subsequent tethering of the FX-Ad5 complex

Liesbeth Lenaerts; Wim van Dam; Leentje Persoons; Lieve Naesens



Development of robust antiviral assays for profiling compounds against a panel of positive-strand RNA viruses using ATP\\/luminescence readout  

Microsoft Academic Search

The development of antiviral assays using an ATP\\/luminescence-based readout to profile antiviral compounds against the positive-strand RNA viruses: yellow fever virus (YFV), West Nile virus (WNV), Sindbis virus, and Coxsackie B virus, representing three virus families, is described. This assay readout is based upon the bioluminescent measurement of ATP in metabolically active cells. Antiviral efficacy was determined by measuring the

Edwin Gong; Tania Ivens; Christel Van den Eynde; Sabine Hallenberger; Kurt Hertogs



Psoriasis Herpeticum due to Varicella Zoster Virus: A Kaposi's Varicelliform Eruption in Erythrodermic Psoriasis  

PubMed Central

Kaposi's varicelliform eruption (KVE) or eczema herpeticum is characterized by disseminated papulovesicular eruption caused by a number of viruses like Herpes simplex virus I and II, Coxsackie virus, and Vaccinia and Small pox viruses in patients with pre-existing skin disease. The occurrence of KVE with psoriasis has been reported recently as a new entity psoriasis herpeticum. The rare causation of psoriasis herpeticum due to Varicella zoster virus in a patient with underlying psoriasis is being reported for the first time.

Garg, Geeta; Thami, Gurvinder P



Coxsackievirus protein 2B modifies endoplasmic reticulum membrane and plasma membrane permeability and facilitates virus release  

Microsoft Academic Search

Digital-imaging microscopy was performed to study the effect of Coxsackie B3 virus infection on the cytosolic free Ca2+ concentration and the Ca2+ content of the endoplasmic reticulum (ER). During the course of infection a gradual increase in the cytosolic free Ca2+ concentration was observed, due to the influx of extracellular Ca2+. The Ca2+ content of the ER decreased in time

Frank J. M. van Kuppeveld; Joost G. J. Hoenderop; Rolf L. L. Smeets; Peter H. G. M. Willems; Henri B. P. M. Dijkman; Jochem M. D. Galama; Willem J. G. Melchers



Complete Genomic Sequencing Shows that Polioviruses and Members of Human Enterovirus Species C Are Closely Related in the Noncapsid Coding Region  

Microsoft Academic Search

The 65 human enterovirus serotypes are currently classified into five species: Poliovirus (3 serotypes), Human enterovirus A (HEV-A) (12 serotypes), HEV-B (37 serotypes), HEV-C (11 serotypes), and HEV-D (2 serotypes). Coxsackie A virus (CAV) serotypes 1, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 24 constitute HEV-C. We have determined the complete genome sequences for the remaining nine

Betty Brown; M. Steven Oberste; Kaija Maher; Mark A. Pallansch



Non-progressive viral myelitis in X-linked agammaglobulinemia  

Microsoft Academic Search

We report a 14-year-old boy with X-linked agammaglobulinemia (XLA) complicated by isolated non-progressive myelitis caused by Coxsackie virus B1. Despite the absence of immunoglobulin supplement and persistence of the virus for the initial 2 years, motor impairment did not show any progression for 3 years. This report shows that the prognosis of central nervous system infection in XLA is not

Kenji Katamura; Haruo Hattori; Tomoko Kunishima; Hirokazu Kanegane; Toshio Miyawaki; Tatsutoshi Nakahata



A six-year study of coxsackievirus B infections in heart disease.  


Virological examination of 385 patients with suspected heart disease and 26 with Bornholm disease over a period of 6 years suggested that Coxsackie group B virus infections were associated with at least half the cases of acute myocarditis and one third of the cases of acute non-bacterial pericarditis. Complement-fixation tests revealed only a few cardiac illnesses associated with other infections (influenza and Mycoplasma pneumoniae). No evidence of infection was found in chronic cardiac disease. PMID:4529604

Grist, N R; Bell, E J



Evaluation of Antiviral and Cytotoxic Activities of Methanolic Extract of Thespesia Populnea (Malvaceae) Flowers  

Microsoft Academic Search

Methanolic flower extracts of Thespesia populnea were evaluated for antiviral and cytotoxic activities using different virus strains cultured in four cell lines. Methanolic extracts showed the strongest antiviral activity against vesicular stomatitis virus, coxsackie B4, and respiratory syncytical viruses that is (EC50 = 20 ?g.mL) and moderate activity for others (EC50 = 100 ?g.mL). These activities may be due to

Saravanakumar Arthanari; P. Renukadevi; J. Vanitha; K. Venkateshwaran; M. Ganesh; E. De Clercq



Genetic analysis of mengovirus protein 2A: its function in polyprotein processing and virus reproduction  

Microsoft Academic Search

To examine the functional requirements of mengo- virus 2A for virus reproduction, a series of mutants with overlapping deletions within the 2A region of mengovirus, and two chimeric constructs in which 2A is replaced either by Theiler's murine encephalo- myelitis virus (TMEV) 2A or by coxsackie B3 virus (CBV3) 2Apro were generated. In vitro polyprotein synthesis showed that in both

Jan Zoll; Frank J. M. van Kuppeveld; Jochem M. D. Galama; Willem J. G. Melchers



de Quervain thyroiditis in a young boy following hand-foot-mouth disease.  


de Quervain thyroiditis, also known as subacute thyroiditis, is a self-limited inflammatory disease of the thyroid gland. It is extremely rare in children. The hallmarks for diagnosis are painful thyroid enlargement, elevated inflammatory markers, and decreased uptake of the thyroid gland on thyroid scintigraphy. Viral infection has been proposed to be associated with de Quervain thyroiditis. Coxsackie virus has been reported to be one of the viruses associated with the disease. To our knowledge, childhood de Quervain thyroiditis associated with hand-foot-mouth disease caused by coxsackie infection has never been reported. We report a 2.7-year-old boy who presented with typical features of de Quervain thyroiditis following hand-foot-mouth disease caused by coxsackie B4 infection. He had a brief thyrotoxic phase initially, followed by transient hypothyroid phase and euthyroidism thereafter. His thyroid scintigraphy showed a typical faint uptake at the diagnosis, and an improvement of the thyroid scan and uptake was shown 8 weeks later. He was treated with prednisolone and nearly complete resolution was documented within 2 months. Careful evaluation of the patient led to the correct diagnosis and appropriate management. PMID:20886354

Engkakul, Pontipa; Mahachoklertwattana, Pat; Poomthavorn, Preamrudee



Psoriasis Herpeticum due to Varicella Zoster Virus: A Kaposi's Varicelliform Eruption in Erythrodermic Psoriasis.  


Kaposi's varicelliform eruption (KVE) or eczema herpeticum is characterized by disseminated papulovesicular eruption caused by a number of viruses like Herpes simplex virus I and II, Coxsackie virus, and Vaccinia and Small pox viruses in patients with pre-existing skin disease. The occurrence of KVE with psoriasis has been reported recently as a new entity psoriasis herpeticum. The rare causation of psoriasis herpeticum due to Varicella zoster virus in a patient with underlying psoriasis is being reported for the first time. PMID:22707775

Garg, Geeta; Thami, Gurvinder P



Infections and dilated cardiomyopathy in Nigeria.  


The relationship of infection to dilated cardiomyopathy is reviewed on the basis of 200 patients seen at University College Hospital, Ibadan. Evidence of infection with Toxoplasma and Coxsackie B viruses is presented. Clinically detectable myocarditis is rare in children, and the preponderance of dilated cardiomyopathy is in patients above the age of 30 years, possibly because there is a long latent period between the initial infection and the development of frank cardiomyopathy. This paper concluded that infections are probably the most important cause of dilated cardiomyopathy in Nigeria. PMID:3038833

Falase, A O



Use of betapropiolactone to disinfect fresh tissue without impairing antigenicity: method applicable to human immunodeficiency virus (HIV) positive tissue.  


A method for inactivating viruses in tissues is reported that does not impair the antigenicity of the Coxsackie virus or of some common tissue antigens, a common problem with standard tissue fixation methods. Tissues can be placed briefly in Betapropiolactone before cryostat sectioning without any adverse effect on preservation or antigen expression. It is suggested that use of Betapropiolactone is applicable to tissues harbouring or exposed to the human immunodeficiency virus (HIV). As betapropiolactone has been reported to be carcinogenic in rodents any potential danger can be avoided by basic simple precautions. PMID:2539401

Chaplin, A J; Heryet, A; Holdsworth, L N; Eglin, R P; Millard, P R



Use of betapropiolactone to disinfect fresh tissue without impairing antigenicity: method applicable to human immunodeficiency virus (HIV) positive tissue.  

PubMed Central

A method for inactivating viruses in tissues is reported that does not impair the antigenicity of the Coxsackie virus or of some common tissue antigens, a common problem with standard tissue fixation methods. Tissues can be placed briefly in Betapropiolactone before cryostat sectioning without any adverse effect on preservation or antigen expression. It is suggested that use of Betapropiolactone is applicable to tissues harbouring or exposed to the human immunodeficiency virus (HIV). As betapropiolactone has been reported to be carcinogenic in rodents any potential danger can be avoided by basic simple precautions. Images Figure

Chaplin, A J; Heryet, A; Holdsworth, L N; Eglin, R P; Millard, P R



Acute right ventricular myocarditis presenting with chest pain and syncope.  


Myocarditis is assumed to involve both ventricles equally. Right ventricular predominant involvement is rarely described. A case of acute viral right ventricular myocarditis presenting with chest pain and syncope, grade 3 atrioventricular block, right ventricular dilatation and free wall hypokinesia is reported. Cardiac MRI showed late enhancement of the right ventricular free wall without involvement of the left ventricle. Anti-Coxsackie A9 virus neutralising IgM-type antibodies titre was elevated. This case emphasises that manifestations of myocarditis can be limited to the right ventricle and should be considered in the differential diagnosis of right ventricular enlargement. PMID:24096068

Mancio, Jennifer; Bettencourt, Nuno; Oliveira, Marco; Pires-Morais, Gustavo; Ribeiro, Vasco Gama



Microorganisms in the aetiology of atherosclerosis  

PubMed Central

Recent publications have suggested that infective pathogens might play an important role in the pathogenesis of atherosclerosis. This review focuses on these microorganisms in the process of atherosclerosis. The results of in vitro studies, animal studies, tissue studies, and serological studies will be summarised, followed by an overall conclusion concerning the strength of the association of the microorganism with the pathogenesis of atherosclerosis. The role of the bacteria Chlamydia pneumoniae and Helicobacter pylori, and the viruses human immunodeficiency virus, coxsackie B virus, cytomegalovirus, Epstein-Barr virus, herpes simplex virus, and measles virus will be discussed. Key Words: atherosclerosis • Chlamydia pneumoniae • Helicobacter pylori

Morre, S; Stooker, W; Lagrand, W; van den Brule, A J C; Niessen, H



Antiviral Spirooliganones A and B with Unprecedented Skeletons from the Roots of Illicium oligandrum.  


Two novel spirooliganones A (1) and (2), a pair of spiro carbon epimers, with a rare dioxaspiro skeleton were isolated from the roots of Illicium oligandrum. The structures were fully determined by spectroscopic analysis and chemical methods, especially modified Mosher's method, and X-ray diffraction analysis. Spirooliganone B was found to exhibit more potent activities against coxsackie virus B3 and influenza virus A (H3N2) (IC50 3.70-5.05 ?M) than spirooliganone A. The biosynthetic pathway involving a hetero-Diels-Alder reaction of the epimers was proposed. PMID:23937631

Ma, Shuang-Gang; Gao, Rong-Mei; Li, Yu-Huan; Jiang, Jian-Dong; Gong, Ning-Bo; Li, Li; Lü, Yang; Tang, Wen-Zhao; Liu, Yun-Bao; Qu, Jing; Lü, Hai-Ning; Li, Yong; Yu, Shi-Shan



Dilated cardiomyopathy in young adult Africans: a sequel to infections?  


Over a period of 6 years, only twelve cases of dilated cardiomyopathy were clinically diagnosed in Nigerians between the ages of 11-30 years at the University College Hospital, Ibadan, Nigeria. Eleven presented with heart failure, while the twelfth patient presented with a cerebrovascular accident. Two other patients also had a cerebrovascular accident. A history of febrile illness was obtained in seven, but in only three was fever unresponsive to antimalarials, documented on admission. Antistreptolysin-O titre was normal and erythrocyte sedimentation rates elevated in each of the patients. Leucocytosis was present in six, three had a four-fold rise or fall in antibody titres against Coxsackie-B viruses and one, a four-fold rise or fall against Toxoplasma gondii. Histological evidences of myopericarditis were found in three of the six patients who died. It is concluded that dilated cardiomyopathy is rare in young adult Nigerians, and that constitutional upset is common, as in children, but prognosis is poorer. Infections by Coxsackie-B viruses, T. gondii and possibly other viruses appear to be of major aetiological factors PMID:6291352

Falase, A O; Sekoni, G A; Adenle, A D



Correlation between virus persistent infection and cardic function in patients with dilated cardiomyopathy.  


In our study, 50 patients with dilated cardiomyopathy (DCM) were selected to investigate the correlation between virus persistent infection and cardic function. We found that 44% of patients with DCM were coxsackie virus B-RNA (CVB-RNA) positive, significantly different from that (20%) of the normal control group (P<0.05). The expression levels of coxsackie adenovirus receptor (CAR) in patients with DCM were significantly higher than those in the normal control group (P<0.01). In CVB-RNA-positive patients, expression levels of CAR were significantly higher than those in CVB-RNA-negative patients (P<0.01). There was a positive correlation between CAR expression and brain natriuretic peptide (BNP) level in patients with DCM, but no significant correlations between the CAR expression level and left ventricular ejection fraction (LVEF) or left ventricular end diastolic diameter (LVEDd). These results showed that expression levels of CAR on the surface of white cells can be used as an indicator for detecting persistent virus infection. We found that expression levels of CAR and heart function in patients with DCM were highly correlated. PMID:23897795

Liu, Qiang; Su, Xiao-jia; Yu, Yan; Liu, Yong-lin



Borna disease virus nucleoprotein inhibits type I interferon induction through the interferon regulatory factor 7 pathway.  


The expression of type I interferon (IFN) is one of the most potent innate defences against viral infection in higher vertebrates. Borna disease virus (BDV) establishes persistent, noncytolytic infections in animals and in cultured cells. Early studies have shown that the BDV phosphoprotein can inhibit the activation of type I IFN through the TBK1-IRF3 pathway. The function of the BDV nucleoprotein in the inhibition of IFN activity is not yet clear. In this study, we demonstrated IRF7 activation and increased IFN-?/? expression in a BDV-persistently infected human oligodendroglia cell line following RNA interference-mediated BDV nucleoprotein silencing. Furthermore, we showed that BDV nucleoprotein prevented the nuclear localisation of IRF7 and inhibited endogenous IFN induction by poly(I:C), coxsackie virus B3 and IFN-?. Our findings provide evidence for a previously undescribed mechanism by which the BDV nucleoprotein inhibits type I IFN expression by interfering with the IRF7 pathway. PMID:23939047

Song, Wuqi; Kao, Wenping; Zhai, Aixia; Qian, Jun; Li, Yujun; Zhang, Qingmeng; Zhao, Hong; Hu, Yunlong; Li, Hui; Zhang, Fengmin



Antiviral activity of Folium isatidis derived extracts in vitro and in vivo.  


Folium isatidis is a native Chinese herbaceous plant widely used for medicinal purposes for thousands of years. However, few studies have focused on the leaves of Isatis indigotica. In this report, we isolated a series of four fractions (I-IV) from Folium isatidis and explored the antiviral activity of each tested extract. The extracts were active against a panel of RNA and DNA viruses in vitro, namely influenza A virus (IAV), coxsackie virus B3 (CVB3), respiratory syncytial virus (RSV), and adenovirus type 7 (Ad-7). Oral administration of 200 mg/kg/d of fraction III in mice exerted strong antiviral effects in viral replication, accompanied by prolonged survival rate, attenuated lung tissue damage as well as significant reductions in pulmonary virus titers and lung index. Our results provide the first biochemical evidence that Folium isatidis and its extracts could be used as potential antiviral agent in the postexposure prophylaxis for multiple viral infections. PMID:23895163

Deng, You-Ping; Liu, Yuan-Yuan; Liu, Zhao; Li, Jin; Zhao, Ling-Min; Xiao, Hong; Ding, Xiao-Hua; Yang, Zhan-Qiu



Cellular immune mechanisms in Coxsackievirus group B, type 3 induced myocarditis in Balb/C mice  

SciTech Connect

Coxsackie B viruses are a common cause of viral myocarditis in humans. A murine model of the human disease has been developed using Coxsackievirus group B, type 3 and inbred Balb/c mice. Infection of T lymphocyte deficient mice does not result in significant myocarditis indicating the importance of T cells in this disease. The virus can be isolated from the hearts of T cell deficient and normal mice in equal concentrations. Virus elimination presumably is mediated by virus specific neutralizing antibody induced in both groups. T lymphocytes, natural killer cells and macrophage obtained from normal virus infected mice are all capable of lysing myofibers in vitro. Maximum lysis is obtained with the cytolytic T cells. When these cell populations or Coxsackievirus immune antibody were adoptively transferred into T lymphocyte deficient animals infected with the virus, only animals given T cells developed significant myocarditis.

Huber, S.A.; Job, L.P.



Epidemic pleurodynia (Bornholm disease) outbreak in Singapore. A clinical and virological study.  


In 1974, an outbreak of Bornholm disease occurred in Singapore. The period 1st May to 31st July was delineated for study. From the clinical presentation 53 patients were placed into two categories "typical Bornholm disease" and "atypical Bornholm disease". The clinical features of only those in the "typical Bornholm disease" group including those with positive Coxsackie B3 virus isolation were described. The virological studies, both faecal isolation for virus and serology were correlated with clinical diagnosis. Fever and characteristic abdominal or chest pain appear to be constant features of Bornholm disease. Positive faecal virus isolation are significantly high in the "typical Bornholm disease" group. Bornholm disease could be diagnosed clinically with fair accuracy. The importance of diagnosing Bornholm disease is emphasized. PMID:1179480

Chong, A Y; Lee, L H; Wong, H B



The Molecular Interaction of CAR and JAML Recruits the Central Cell Signal Transducer PI3K  

SciTech Connect

Coxsackie and adenovirus receptor (CAR) is the primary cellular receptor for group B coxsackieviruses and most adenovirus serotypes and plays a crucial role in adenoviral gene therapy. Recent discovery of the interaction between junctional adhesion molecule-like protein (JAML) and CAR uncovered important functional roles in immunity, inflammation, and tissue homeostasis. Crystal structures of JAML ectodomain (2.2 angstroms) and its complex with CAR (2.8 angstroms) reveal an unusual immunoglobulin-domain assembly for JAML and a charged interface that confers high specificity. Biochemical and mutagenesis studies illustrate how CAR-mediated clustering of JAML recruits phosphoinositide 3-kinase (P13K) to a JAML intracellular sequence motif as delineated for the {alpha}{beta} T cell costimulatory receptor CD28. Thus, CAR and JAML are cell signaling receptors of the immune system with implications for asthma, cancer, and chronic nonhealing wounds.

Verdino, Petra; Witherden, Deborah A.; Havran, Wendy L.; Wilson, Ian A. (Scripps)



Impact of the coxsackievirus and adenovirus receptor on the adenoma-carcinoma sequence of colon cancer  

PubMed Central

Background: Coxsackie and adenovirus receptor (CAR) has been suggested to function as a tumour suppressor. Its impact on the adenoma–carcinoma sequence of the colon, however, is unclear. Methods: Coxsackie and adenovirus receptor was analysed in non-cancerous and neoplastic colon samples using immunohistochemistry and quantitative RT–PCR. The function of CAR in colon cancer cell lines was determined following application of CAR siRNA or ectopic expression of a human full-length CAR cDNA. Results: Compared with healthy mucosa, increased CAR-mRNA expression was found in adenomas, whereas primary cancers and metastases displayed a marked decline. At the plasma membrane, CAR was present in normal mucosa samples (93%), adenomas, and metastases (100% ea.), whereas in colon cancers, it was found less frequently (49%, P<0.0001). Cytoplasmic CAR immunopositivity increased from normal mucosa (22%), to adenomas (73%, P=0.0006), primary cancers (83%, P<0.0001), and metastases (67%, P=0.0019). In cancer cell lines, CAR inhibition resulted in increased proliferation, whereas enforced ectopic CAR expression led to opposite results. Blocking the extracellular portion of CAR increased cell invasion in vitro. In mice, xenotransplants of colon cancer cells with enforced CAR expression formed significantly smaller tumours, whereas CAR inhibition increased the formation of liver metastases. Conclusion: We conclude that CAR facilitates complex effects during colon carcinogenesis, potentially mediated by its stage-dependent subcellular distribution; high CAR expression potentially prevents apoptosis in adenomas, loss of CAR at the plasma membrane promotes growth, and dissemination of primary cancers, and high membranous CAR presence may support the establishment of distant metastases.

Stecker, K; Vieth, M; Koschel, A; Wiedenmann, B; Rocken, C; Anders, M



Co-circulation and evolution of polioviruses and species C enteroviruses in a district of Madagascar.  


Between October 2001 and April 2002, five cases of acute flaccid paralysis (AFP) associated with type 2 vaccine-derived polioviruses (VDPVs) were reported in the southern province of the Republic of Madagascar. To determine viral factors that favor the emergence of these pathogenic VDPVs, we analyzed in detail their genomic and phenotypic characteristics and compared them with co-circulating enteroviruses. These VDPVs appeared to belong to two independent recombinant lineages with sequences from the type 2 strain of the oral poliovaccine (OPV) in the 5'-half of the genome and sequences derived from unidentified species C enteroviruses (HEV-C) in the 3'-half. VDPV strains showed characteristics similar to those of wild neurovirulent viruses including neurovirulence in poliovirus-receptor transgenic mice. We looked for other VDPVs and for circulating enteroviruses in 316 stools collected from healthy children living in the small area where most of the AFP cases occurred. We found vaccine PVs, two VDPVs similar to those found in AFP cases, some echoviruses, and above all, many serotypes of coxsackie A viruses belonging to HEV-C, with substantial genetic diversity. Several coxsackie viruses A17 and A13 carried nucleotide sequences closely related to the 2C and the 3D(pol) coding regions of the VDPVs, respectively. There was also evidence of multiple genetic recombination events among the HEV-C resulting in numerous recombinant genotypes. This indicates that co-circulation of HEV-C and OPV strains is associated with evolution by recombination, resulting in unexpectedly extensive viral diversity in small human populations in some tropical regions. This probably contributed to the emergence of recombinant VDPVs. These findings give further insight into viral ecosystems and the evolutionary processes that shape viral biodiversity. PMID:18085822

Rakoto-Andrianarivelo, Mala; Guillot, Sophie; Iber, Jane; Balanant, Jean; Blondel, Bruno; Riquet, Franck; Martin, Javier; Kew, Olen; Randriamanalina, Bakolalao; Razafinimpiasa, Lalatiana; Rousset, Dominique; Delpeyroux, Francis



Structural basis for antiviral inhibition of the main protease, 3C, from human enterovirus 93.  


Members of the Enterovirus genus of the Picornaviridae family are abundant, with common human pathogens that belong to the rhinovirus (HRV) and enterovirus (EV) species, including diverse echo-, coxsackie- and polioviruses. They cause a wide spectrum of clinical manifestations ranging from asymptomatic to severe diseases with neurological and/or cardiac manifestations. Pandemic outbreaks of EVs may be accompanied by meningitis and/or paralysis and can be fatal. However, no effective prophylaxis or antiviral treatment against most EVs is available. The EV RNA genome directs the synthesis of a single polyprotein that is autocatalytically processed into mature proteins at Gln?Gly cleavage sites by the 3C protease (3C(pro)), which has narrow, conserved substrate specificity. These cleavages are essential for virus replication, making 3C(pro) an excellent target for antivirus drug development. In this study, we report the first determination of the crystal structure of 3C(pro) from an enterovirus B, EV-93, a recently identified pathogen, alone and in complex with the anti-HRV molecules compound 1 (AG7404) and rupintrivir (AG7088) at resolutions of 1.9, 1.3, and 1.5 Å, respectively. The EV-93 3C(pro) adopts a chymotrypsin-like fold with a canonically configured oxyanion hole and a substrate binding pocket similar to that of rhino-, coxsackie- and poliovirus 3C proteases. We show that compound 1 and rupintrivir are both active against EV-93 in infected cells and inhibit the proteolytic activity of EV-93 3C(pro) in vitro. These results provide a framework for further structure-guided optimization of the tested compounds to produce antiviral drugs against a broad range of EV species. PMID:21835784

Costenaro, Lionel; Kaczmarska, Zuzanna; Arnan, Carme; Janowski, Robert; Coutard, Bruno; Solà, Maria; Gorbalenya, Alexander E; Norder, Heléne; Canard, Bruno; Coll, Miquel



Diagnostic relevance of humoral and cell-mediated immune reactions in patients with acute viral myocarditis.  

PubMed Central

Sera of 177 patients with acute myocarditis (10 coxsackie B 3/4, four influenza, four mumps, 15 cytomegalovirus, 144 undefined) were tested by indirect immunofluorescence for autoantibodies against heart and skeletal muscle and vital or air-dried adult cardiocytes. Antibody-dependent cytolysis, lymphocytotoxicity and antibody-dependent cellular lymphocytotoxicity were assessed using viral adult rat cardiocytes as target cells. Muscle-specific anti-sarcolemmal antibodies of the anti-myolemmal type--often associated with non-organ-specific anti-endothelial antibodies--were demonstrated in nine out of 10 patients with coxsackie B, in all patients with influenza and mumps and in 65 out of 144 patients with undefined myocarditis. In contrast, 13 out of 15 patients with cytomegalovirus myocarditis lacked anti-sarcolemmal antibodies but had low titre anti-inter fibrillary antibodies instead. In the presence of complement, anti-myolemmal antibodies induced cytolysis of vital cardiocytes, whereas hepatocytes remained unaffected. Titres of anti-myolemmal antibodies correlated with the degree of cardiocytolysis. The anti-myolemmal immunofluorescent pattern and the cytolytic serum activity could be absorbed with the respective viral antigens suggesting that these antibodies cross-react with moieties of the virus itself and may be both diagnostic and aetiological markers in acute viral myocarditis. Lymphocyte-mediated cytotoxicity against heterologous cardiac target cells could not be observed in our patients with myocarditis of proven viral aetiology. However, lymphocyte-mediated cytotoxicity was demonstrated in 10 ASA-positive and one ASA-negative patient with myocarditis of unknown origin. ASA-positive sera blocked lymphocytotoxicity in three of these patients.

Maisch, B; Trostel-Soeder, R; Stechemesser, E; Berg, P A; Kochsiek, K



Detection of Viral Pathogens by Reverse Transcriptase PCR and of Microbial Indicators by Standard Methods in the Canals of the Florida Keys  

PubMed Central

In order to assess the microbial water quality in canal waters throughout the Florida Keys, a survey was conducted to determine the concentration of microbial fecal indicators and the presence of human pathogenic microorganisms. A total of 19 sites, including 17 canal sites and 2 nearshore water sites, were assayed for total coliforms, fecal coliforms, Escherichia coli, Clostridium perfringens, enterococci, coliphages, F-specific (F+) RNA coliphages, Giardia lamblia, Cryptosporidium parvum, and human enteric viruses (polioviruses, coxsackie A and B viruses, echoviruses, hepatitis A viruses, Norwalk viruses, and small round-structured viruses). Numbers of coliforms ranged from <1 to 1,410, E. coli organisms from <1 to 130, Clostridium spp. from <1 to 520, and enterococci from <1 to 800 CFU/100 ml of sample. Two sites were positive for coliphages, but no F+ phages were identified. The sites were ranked according to microbial water quality and compared to various water quality standards and guidelines. Seventy-nine percent of the sites were positive for the presence of enteroviruses by reverse transcriptase PCR (polioviruses, coxsackie A and B viruses, and echoviruses). Sixty-three percent of the sites were positive for the presence of hepatitis A viruses. Ten percent of the sites were positive for the presence of Norwalk viruses. Ninety-five percent of the sites were positive for at least one of the virus groups. These results indicate that the canals and nearshore waters throughout the Florida Keys are being impacted by human fecal material carrying human enteric viruses through current wastewater treatment strategies such as septic tanks. Exposure to canal waters through recreation and work may be contributing to human health risks.

Griffin, Dale W.; Gibson, Charles J.; Lipp, Erin K.; Riley, Kelley; Paul, John H.; Rose, Joan B.



Structure of the human adenovirus serotype 2 fiber head domain at 1.5 A resolution.  


Adenovirus binds to its receptor via the head domain of its fiber protein. We have crystallized the adenovirus serotype 2 (subgroup C) receptor binding domain and solved the structure at 1.5 A resolution by the molecular replacement technique using the known adenovirus type 5 head structure. Included in the high-resolution model are 306 water molecules, five alternative side chain conformations, and individual anisotropic temperature factors for each atom. The overall structure of the serotype 2 head is very similar to its serotype 5 homologue, apart from differences in some of the flexible loops. All but subgroup B adenoviruses are believed to use the recently identified protein CAR (Coxsackievirus and adenovirus receptor) as receptor. By comparison of the two structures and sequence alignment of CAR binding and non-CAR binding serotype fiber heads, we discuss possible receptor binding sites and propose a receptor binding site in a crevice between two monomers on the side of the trimer. The structural basis of the extraordinary stability of the fiber head trimer is also discussed. PMID:10502512

van Raaij, M J; Louis, N; Chroboczek, J; Cusack, S



A non-oncogenic HPV 16 E6/E7 vaccine enhances treatment of HPV expressing tumors.  


Human papillomaviruses (HPVs) are the causative factor for >90% of cervical cancers and 25% of head and neck cancers. The incidence of HPV positive (+) head and neck squamous cell carcinomas has greatly increased in the last 30 years. E6 and E7 are the two key viral oncoproteins that induce and propagate cellular transformation. An immune response generated during cisplatin/radiation therapy improves tumor clearance of HPV(+) cancers. Augmenting this induced response during therapy with an adenoviral HPV16 E6/E7 vaccine improves long-term survival in pre-clinical models. Here, we describe the generation of an HPV16 E6/E7 construct, which contains mutations that render E6/E7 non-oncogenic, while preserving antigenicity. These mutations do not allow E6/E7 to degrade p53, pRb, PTPN13, or activate telomerase. Non-oncogenic E6/E7 (E6(?)/E7(?)) expressed as a stable integrant, or in the [E1-, E2b-] adenovirus, lacks the ability to transform human cells while retaining the ability to induce an HPV-specific immune response. Moreover, E6(?)/E7(?) plus chemotherapy/radiation statistically enhances clearance of established HPV(+) cancer in vivo. PMID:22918471

Wieking, B G; Vermeer, D W; Spanos, W C; Lee, K M; Vermeer, P; Lee, W T; Xu, Y; Gabitzsch, E S; Balcaitis, S; Balint, J P; Jones, F R; Lee, J H



Identification of a novel HLA-A*24:02-restricted adenovirus serotype 11-specific CD8(+) T-cell epitope for adoptive immunotherapy.  


Subgroup B adenovirus serotype 11 (Ad11) occasionally causes fatal infections in immunocompromised patients. The present study describes a novel Ad11 epitope presented by HLA-A*24:02 that could be used for adoptive immunotherapy. Ten synthetic Ad11 hexon protein-derived nonamer peptides that bound to HLA-A*24:02 were selected by a computer algorithm and MHC stabilization assay. Stimulation of peripheral blood mononuclear cells from HLA-A*24:02+ donors with each of these synthetic peptides induced peptide-specific CD8(+) T-cells for three peptides. Testing the reactivity of these peptide-specific CD8(+) T-cells against various target cells confirmed that peptide TYFNLGNKF is naturally processed in Ad11-infected cells and is presented by HLA-A*24:02. Emergence of TYFNLGNKF-specific CD8(+) T-cells coincided with the clearance of adenoviruses in a patient with Ad11 disease. Importantly, TYFNLGNKF-specific CD8(+) T-cells were suggested to be not serotype cross-reactive. The novel HLA-A*24:02-restricted Ad11 epitope could be used for anti-Ad11 adoptive immunotherapy and to monitor immunity to Ad11 using MHC tetramers. PMID:23911395

Imahashi, Nobuhiko; Nishida, Tetsuya; Ito, Yoshinori; Kawada, Jun-Ichi; Nakazawa, Yozo; Toji, Shingo; Suzuki, Susumu; Terakura, Seitaro; Kato, Tomonori; Murata, Makoto; Naoe, Tomoki




PubMed Central

Chromatographic behavior of whole type 1 poliovirus and phenol-extracted viral RNA on diethylaminoethyl cellulose columns, as revealed by assay of plaque-forming capacity, indicated that infectious RNA had surface properties markedly different from those of the intact virus. Infectious RNA of type 1 poliovirus and Coxsackie B1 virus was relatively resistant to heat inactivation as compared to intact virus. Kinetics of inactivation at elevated temperatures were multi-hit in character. The structure of infectious enterovirus RNA was investigated by treatment with chemical inactivating agents. Urea and guanidine as hydrogen bond-disrupting agents, and mercaptoethanol and thioglycolate as disulfide bond-disrupting agents, and combinations of these did not destroy RNA infectivity whereas hydrogen bond-disrupting treatment inactivated intact virus rapidly. RNA infectivity was not reduced by chloroform extraction alone, or by octanol extraction alone, but was reduced by chloroform-octanol extraction which failed to depolymerize RNA to an extent detectable by ultracentrifugal analysis. Infectivity of type 1 poliovirus and Coxsackie B1 virus RNA was destroyed in accordance with first order kinetics by very dilute solutions of pancreatic ribonuclease, and by purified snake venom phosphodiesterase, but not at all by bacterial alkaline phosphatase. Inactivation by venom diesterase was not accelerated by prior treatment of RNA with bacterial alkaline phosphatase. These results indicated that infectivity of enteroviral RNA resided in a single stranded structure, that a single break of a phosphodiester bond anywhere along the structure was sufficient to destroy infectivity, and that infectivity did not require a terminal phosphate group. Hydroxylamine, but not other carbonyl reagents, rapidly destroyed infectivity of intact type 1 poliovirus viral RNA without depolymerization of RNA-detectable by behavior in the analytical ultracentrifuge. With S35-methionine-labeled poliovirus a very small fraction of radioactivity remained in RNA preparations following phenol extraction. No evidence could be obtained to indicate that infectious enteroviral RNA was composed of subunits. RNA extracted with phenol during the course of infection of HeLa cells with type 1 poliovirus resembled RNA obtained from purified whole virus with respect to heat inactivation, hydroxylamine inactivation, chromatographic separation, susceptibility to protein denaturing agents, and ability to infect productively both naturally susceptible HeLa cells and naturally insusceptible L strain mouse cells. Intracellular production of infectious RNA paralleled intracellular maturation of whole virus and preceded it by a very short interval.

Holland, John J.; McLaren, Leroy C.; Hoyer, Bill H.; Syverton, Jerome T.



Nuclear factor kappa B is involved in lipopolysaccharide-stimulated induction of interferon regulatory factor-1 and GAS/GAF DNA-binding in human umbilical vein endothelial cells  

PubMed Central

In this study we examined the signalling events that regulate lipopolysaccharide (LPS)-stimulated induction of interferon regulatory factor (IRF)-1 in human umbilical vein endothelial cells (HUVECs). LPS stimulated a time- and concentration-dependent increase in IRF-1 protein expression, an effect that was mimicked by the cytokine, tumour necrosis factor (TNF)-?. LPS stimulated a rapid increase in nuclear factor kappa B (NF?B) DNA-binding activity. Pre-incubation with the NF?B pathway inhibitors, N-?-tosyl-L-lysine chloromethyl ketone (TLCK) or pyrrolidine dithiocarbamate (PDTC), or infection with adenovirus encoding I?B?, blocked both IRF-1 induction and NF?B DNA-binding activity. LPS and TNF? also stimulated a rapid activation of gamma interferon activation site/gamma interferon activation factor (GAS/GAF) DNA-binding in HUVECs. Preincubation with the Janus kinase (JAK)-2 inhibitor, AG490 blocked LPS-stimulated IRF-1 induction but did not affect GAS/GAF DNA-binding. Preincubation with TLCK, PDTC or infection with I?B? adenovirus abolished LPS-stimulated GAS/GAF DNA-binding. Incubation of nuclear extracts with antibodies to RelA/p50 supershifted GAS/GAF DNA-binding demonstrating the involvement of NF?B isoforms in the formation of the GAS/GAF complex. These studies show that NF?B plays an important role in the regulation of IRF-1 induction in HUVECs. This is in part due to the interaction of NF?B isoforms with the GAS/GAF complex either directly or via an intermediate protein.

Liu, Li; Paul, Andrew; MacKenzie, Christopher J; Bryant, Clare; Graham, Anne; Plevin, Robin



Functional diversity of E1A gene autoregulation among human adenoviruses.  

PubMed Central

Autoregulation of the adenovirus E1A gene involves its constitutive expression and positively and negatively regulated transcription. Dissection of this process will identify basal-level cis elements and autoregulatory targets of the E1A promoter and functional domains within the trans-acting E1A gene products. In this report, the DNA sequence of the human subgroup B adenovirus type 3 (Ad3) E1A gene is presented and compared with that of the E1A genes of similar and distantly related human adenoviruses. The cDNA forms of the Ad3 E1A gene, corresponding to two major early mRNA species, are cloned, sequenced, and subcloned into plasmid expression vectors. Cotransfections of cell cultures are performed with Ad5 or Ad3 E1A gene expression plasmids and a reporter gene under control of the Ad5 or Ad3 E1A promoter. The Ad5 and Ad3 E1A promoters are similarly repressed by either serotype's 12S cDNA gene products. The Ad3 E1A promoter responds much more strongly than the Ad5 E1A promoter to transactivation by 13S cDNA gene products. In contrast, the 13S cDNA gene of Ad5 has greater transactivation activity than that of Ad3. Experiments with missense mutations of the Ad5 E1A gene indicate that transactivation of the Ad5 E1A promoter is weak, just reversing or balancing negative autorepression. Single amino acid substitutions in the conserved, repressive functional domain 2 of the E1A gene modulate transactivating activity that is usually associated with the separate and distal conserved functional domain 3. These results suggest a strong structure-function relationship influenced by the variable sequences separating these conserved domains. Images

Cogan, J D; Jones, S N; Hall, R K; Tibbetts, C



Vascular endothelial growth factor signalling in endothelial cell survival: A role for NF{kappa}B  

SciTech Connect

Angiogenesis is the development of blood capillaries from pre-existing vessels. Vascular endothelial growth factor (VEGF) is a key regulator of vessel growth and regression, and acts as an endothelial survival factor by protecting endothelial cells from apoptosis. Many genes involved in cell proliferation and apoptosis are regulated by the nuclear factor kappa B (NF{kappa}B) transcription factor family. This study aimed to address the hypothesis that VEGF-mediated survival effects on endothelium involve NF{kappa}B. Using an NF{kappa}B-luciferase reporter adenovirus, we observed activation of NF{kappa}B following VEGF treatment of human umbilical vein endothelial cells. This was confirmed using electrophoretic mobility shift assay and found to involve nuclear translocation of NF{kappa}B sub-unit p65. However, NF{kappa}B activation occurred without degradation of inhibitory I{kappa}B proteins (I{kappa}B{alpha}, I{kappa}B{beta}, and I{kappa}B{epsilon}). Instead, tyrosine phosphorylation of I{kappa}B{alpha} was observed following VEGF treatment, suggesting NF{kappa}B activation was mediated by degradation-independent dissociation of I{kappa}B{alpha} from NF{kappa}B. Adenovirus-mediated over-expression of either native I{kappa}B{alpha}, or of I{kappa}B{alpha} in which tyrosine residue 42 was mutated to phenylalanine, inhibited induction of NF{kappa}B-dependent luciferase activity in response to VEGF. Furthermore, VEGF-induced upregulation of mRNA for the anti-apoptotic protein Bcl-2 and cell survival following serum withdrawal was reduced following I{kappa}B{alpha} over-expression. This study highlights that different molecular mechanisms of NF{kappa}B activation may be involved downstream of stimuli which activate the endothelial lining of blood vessels.

Grosjean, Jennifer [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom)]. E-mail:; Kiriakidis, Serafim [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom); Reilly, Kerri [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom); Feldmann, Marc [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom); Paleolog, Ewa [Kennedy Institute of Rheumatology, Faculty of Medicine, Imperial College, London (United Kingdom)



Oxidative stress in coronary artery disease: epigenetic perspective.  


The association between oxidative stress and coronary artery disease (CAD) is well documented. However, the role of epigenetic factors contributing to oxidative stress is relatively unexplored. In this study, we aimed to explore the impact of DNA methylation profile in BCL2/E1B adenovirus interacting protein 3 (BNIP3), extracellular superoxide dismutase (EC-SOD) and glutathione-S-transferase P1 (GSTP1) on the oxidative stress in CAD. Further, the contribution of folate pathway genetic polymorphisms in regulating epigenome was elucidated. The expression of BNIP3, EC-SOD, and GSTP1 were studied by using Maxima@SYBR-green based real-time qPCR approach in peripheral blood samples. Combined bisulfite restriction analysis and methylation-specific PCR were used to study promoter CpG island methylation. Further, the effect of homocysteine on BNIP3 gene expression was studied in human aortic endothelial cells in vitro. CAD cases exhibited upregulation of BNIP3, downregulation of EC-SOD and GSTP1. Hypomethylation of BNIP3 and hypermethylation of EC-SOD were observed in CAD cases. The expression of BNIP3 was positively correlated with homocysteine, MDA, protein carbonyls, and methylene tetrahydrofolate reductase C677T, while showing inverse association with cytosolic serine hydroxymethyl transferase C1420T. The expressions of EC-SOD and GSTP1 showed positive association with thymidylate synthase (TYMS) 2R3R, while inverse association with MDA, protein carbonyls, and methionine synthase reductase (MTRR) A66G. In vitro analysis showed homocysteine-dependent upregulation of BNIP3. The results of this study suggest that the aberrations in one-carbon metabolism appear to induce altered gene expression of EC-SOD, GSTP1, and BNIP3, and thus contribute to the increased oxidative stress and increased susceptibility to CAD. PMID:23160801

Lakshmi, Sana Venkata Vijaya; Naushad, Shaik Mohammad; Reddy, Cheruku Apoorva; Saumya, Kankanala; Rao, Damera Seshagiri; Kotamraju, Srigiridhar; Kutala, Vijay Kumar



Protein aggregation during overexpression limited by peptide extensions with large net negative charge.  


Folding of the human coxsackie and adenovirus receptor immunoglobulin (Ig) variable-type domain (CAR D1) during overexpression in the Escherichia coli cytoplasm was shown previously to be partially rescued by fusion to a 22-residue C-terminal peptide. Here, peptide sequence features required for solubilization and folding of CAR D1 and similar Ig variable-type domains from two other human membrane proteins were investigated. Peptide extensions with net negative charge > -6 fully solubilized CAR D1, and approximately half of the peptide-solubilized protein was correctly folded. The Ig variable-type domains from human A33 antigen and myelin P-zero proteins were only partially solubilized by peptide extensions with net charge of -12, however, and only the solubilized P-zero domain appeared to fold correctly whereas the A33 domain formed soluble microaggregates of misfolded protein. Our results suggest a model where the large net charge of peptide extensions increases electrostatic repulsion between nascent polypeptides. The resulting decrease in aggregation rate can enable some polypeptides to fold spontaneously into their native protein conformations. Analysis of the solubility and folding status of sets of structurally homologous proteins, such as the Ig variable-type domains described here, during overexpression could provide insights into how amino acid and gene sequences influence the efficiency of spontaneous protein folding. PMID:15249042

Zhang, Yian-Biao; Howitt, Jason; McCorkle, Sean; Lawrence, Paul; Springer, Karen; Freimuth, Paul



Echovirus 22 is an atypical enterovirus.  

PubMed Central

Although echovirus 22 (EV22) is classified as an enterovirus in the family Picornaviridae, it is atypical of the enterovirus paradigm, typified by the polioviruses and the coxsackie B viruses. cDNA reverse transcribed from coxsackievirus B3 (CVB3) RNA does not hybridize to genomic RNA of EV22, and conversely, cDNA made to EV22 does not hybridize to CVB3 genomic RNA or to molecular clones of CVB3 or poliovirus type 1. EV22 cDNA does not hybridize to viral RNA of encephalomyocarditis virus or to a molecular clone of Theiler's murine encephalomyelitis virus, members of the cardiovirus genus. The genomic RNA of EV22 cannot be detected by the polymerase chain reaction using generic enteroviral primers. EV22 does not shut off host cell protein synthesis, and the RNA of EV22 is efficiently translated in vitro in rabbit reticulocyte lysates. Murine enterovirus-immune T cells recognize and proliferate against EV22 as an antigen in vitro, demonstrating that EV22 shares an epitope(s) common to enteroviruses but not found among other picornaviruses. Images

Coller, B A; Chapman, N M; Beck, M A; Pallansch, M A; Gauntt, C J; Tracy, S M



Systemically administered liposome-encapsulated Ad-PEDF potentiates the anti-cancer effects in mouse lung metastasis melanoma  

PubMed Central

Background The use of adenoviral vector for gene therapy is still an important strategy for advanced cancers, however, the lack of the requisite coxsackie-adenovirus receptor in cancer cells and host immune response to adenovirus limit the application of adenoviral vector in vivo. Method We designed the antiangiogenic gene therapy with recombinant PEDF adenovirus (Ad-PEDF) encapsulated in cationic liposome (Ad-PEDF/Liposome), and investigated the anti-tumor efficacy of Ad-PEDF/Liposome complex on inhibition of tumor metastasis. Results We found that systemic administration of Ad-PEDF/liposome was well tolerated and resulted in marked suppression of tumor growth, and was more potent than uncoated Ad-PEDF to induce apoptosis in B16-F10 melanoma cells and inhibit murine pulmonary metastases in vivo. After Ad-luciferase was encapsulated with liposome, its distribution decreased in liver and increased in lung. The anti-Ad IgG level of Ad-PEDF/Liposome was significantly lower than Ad-PEDF used alone. Conclusion The present findings provide evidences of systematic administration of cationic liposome-encapsulated Ad-PEDF in pulmonary metastatic melanoma mice model, and show an encouraging therapeutic effect for further exploration and application of more complexes based on liposome-encapsulated adenovirus for more cancers.



Nasal cardiac myosin peptide treatment and OX40 blockade protect mice from acute and chronic virally-induced myocarditis  

PubMed Central

Myocarditis poses a severe health problem, can lead to dilated cardiomyopathy (DCM) and death, and is thought to be triggered by infections. Enteroviruses such as Coxsackie virus B 3 (CVB3) have been implicated as a culprit, since they can cause acute and chronic heart disease in susceptible mice. CVB was detected in human cardiac myocytes in some cases, whereas acute CVB infection was thought to have caused death. Here we studied, whether nasal administration of cardiac myosin (CM) major histocompatibility class (MHC) II peptides CM947-960 and CM735-747 and OX40 blockade would be able to ameliorate immunopathology and heart disease in BALB/C mice infected with CVB3. We found that nasal CM peptide prophylactic treatment significantly reduced myocarditis and mortality by enhancing Treg and IL-10 induction and that blockade of OX40 signaling could reduce heart inflammation when administered late during pathogenesis. Altogether, these results chart the way for novel prevention and intervention strategies for viral myocarditis.

Fousteri, Georgia; Dave, Amy; Morin, Bret; Omid, Shaida; Croft, Michael; von Herrath, Matthias G



Non-classical export of an adenovirus structural protein.  


The icosahedral capsids of Adenoviruses (Ads) consist of the hexon and stabilizing proteins building the facettes, and of the vertex protein penton base (Pb) anchoring the protruding fibers. The fibers bind to the Coxsackie virus B Ad cell surface receptor (CAR) and Pb to integrins. Here we describe a novel property of the Ad2 Pb. Pb was found to leave the infected cell and, upon exit, it attached to the surrounding noninfected cells forming a radial gradient with highest Pb levels on cells adjacent to the infected cell. The producer cells remained intact until at least 30 h post infection. At this point, Pb was not recovered from the extracellular medium, suggesting that its cell-cell spread might not involve free Pb. When viral particles were released at late stages of infection, soluble Pb was found in the extracellular medium and it randomly bound to noninfected cells. Nonlytic export of Pb occurred upon transient transfection with plasmid DNA, but plasmid-encoded fiber was not exported, indicating that cell-cell spread of Pb is autonomous of infection. Pb export was not affected by Brefeldin A-induced disruption of the Golgi apparatus, suggesting that it occurred via a nonclassical mechanism. Interestingly, the coexpression of Pb and fiber leads to both Pb and fiber export, termed 'protein abduction'. We suggest that fiber abduction might support viral dissemination in infected tissues by interfering with tissue integrity. PMID:12753648

Trotman, Lloyd C; Achermann, Dominik P; Keller, Stephan; Straub, Monika; Greber, Urs F



On the need for, and the delivery of, cross-protective vaccines.  


The rhinoviruses that are instrumental in causing about one-third of the outbreaks of the common cold present us with some 100 or so serotypes whose convalescent sera do not cross-neutralise. A similar situation prevails with the organism that causes gonorrhoea. Both the HIV and the protozoan causing malaria are notorious for their ability to evade the immune system by changes to their antigenic profile. Similarly, we face continual changes in the antigenic determinants of the influenza virus. It is clear that we require vaccine for these diseases that provide protection against a wide variety of basic variants. This can be achieved, as was shown by Arvind Kumar, who, in his PhD project, generated monoclonal antibodies to cross-reacting yet neutralising epitopes of a number of rhinoviruses. Such antibodies also neutralised some Coxsackie viruses as well as some of the types of Poliovirus. This demonstration of feasibility will be explored further in my paper with a view to arriving at a general approach to the production of vaccines whose humoral and cellular responses can neutralise a wide cross-section of serotype variants. PMID:15755565

Spier, R E



Rapid and highly sensitive detection of Enterovirus 71 by using nanogold-enhanced electrochemical impedance spectroscopy  

NASA Astrophysics Data System (ADS)

Enterovirus 71 (EV71) infection is an emerging infectious disease causing neurological complications and/or death within two to three days after the development of fever and rash. A low viral titre in clinical specimens makes the detection of EV71 difficult. Conventional approaches for detecting EV71 are time consuming, poorly sensitive, or complicated, and cannot be used effectively for clinical diagnosis. Furthermore, EV71 and Coxsackie virus A16 (CA16) may cross react in conventional assays. Therefore, a rapid, highly sensitive, specific, and user-friendly test is needed. We developed an EV71-specific nanogold-modified working electrode for electrochemical impedance spectroscopy in the detection of EV71. Our results show that EV71 can be distinguished from CA16, Herpes simplex virus, and lysozyme, with the modified nanogold electrode being able to detect EV71 in concentrations as low as 1 copy number/50 ?l reaction volume, and the duration between sample preparation and detection being 11 min. This detection platform may have the potential for use in point-of-care diagnostics.

Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Lu, Yu-Ning; Wang, Fang-Yu; Tsai, Li-Yun; Shieh, Juo-Yu; Yang, Jyh-Yuan; Juan, Chien-Chang; Tu, Lung-Chen; Chang, Chia-Ching



High sensitivity and label-free detection of Enterovirus 71 by nanogold modified electrochemical impedance spectroscopy  

NASA Astrophysics Data System (ADS)

Enterovirus 71 (EV71), which is the most fulminant and invasive species of enterovirus, can cause children neurologic complications and death within 2-3 days after fever and rash developed. Besides, EV71 has high sequence similarity with Coxsackie A 16 (CA16) that makes differential diagnosis difficult in clinic and laboratory. Since conventional viral diagnostic method cannot diagnose EV71 quickly and EV71 can transmit at low viral titer, the patients might delay in treatment. A quick, high sensitive, and high specific test for EV71 detection is pivotal. Electrochemical impedance spectroscopy (EIS) has been applied for detecting bio-molecules as biosensors recently. In this study, we try to build a detection platform for EV71 detection by nanogold modified EIS probe. The result shows that our probe can detect 3.6 VP1/50 ?l (one EV71 particle has 60 VP1) in 3 minutes. The test can also distinguish EV71 from CA16 and lysozyme. Diagnosis of enterovirus 71 by electrochemical impedance spectroscopy has the potential to apply in clinic.

Wang, Fang-Yu; Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Yang, Jyh-Yuan; Chang, Chia-Ching



Adenovirus Gene Transfer to Amelogenesis Imperfecta Ameloblast-Like Cells  

PubMed Central

To explore gene therapy strategies for amelogenesis imperfecta (AI), a human ameloblast-like cell population was established from third molars of an AI-affected patient. These cells were characterized by expression of cytokeratin 14, major enamel proteins and alkaline phosphatase staining. Suboptimal transduction of the ameloblast-like cells by an adenovirus type 5 (Ad5) vector was consistent with lower levels of the coxsackie-and-adenovirus receptor (CAR) on those cells relative to CAR-positive A549 cells. To overcome CAR -deficiency, we evaluated capsid-modified Ad5 vectors with various genetic capsid modifications including “pK7” and/or “RGD” motif-containing short peptides incorporated in the capsid protein fiber as well as fiber chimera with the Ad serotype 3 (Ad3) fiber “knob” domain. All fiber modifications provided an augmented transduction of AI-ameloblasts, revealed following vector dose normalization in A549 cells with a superior effect (up to 404-fold) of pK7/RGD double modification. This robust infectivity enhancement occurred through vector binding to both ?v?3/?v?5 integrins and heparan sulfate proteoglycans (HSPGs) highly expressed by AI-ameloblasts as revealed by gene transfer blocking experiments. This work thus not only pioneers establishment of human AI ameloblast-like cell population as a model for in vitro studies but also reveals an optimal infectivity-enhancement strategy for a potential Ad5 vector-mediated gene therapy for AI.

Borovjagin, Anton V.; Ren, Changchun; Lamani, Ejvis; Mamaeva, Olga A.; Wu, Hongju; Keyser, Enid; Murakami, Miho; Chen, Shuo



Presence of the Coxsackievirus and Adenovirus Receptor (CAR) in human neoplasms: a multitumour array analysis.  


Background:The Coxsackie- and Adenovirus Receptor (CAR) has been assigned two crucial attributes in carcinomas: (a) involvement in the regulation of growth and dissemination and (b) binding for potentially therapeutic adenoviruses. However, data on CAR expression in cancer types are conflicting and several entities have not been analysed to date.Methods:The expression of CAR was assessed by immunohistochemical staining of tissue microarrays (TMA) containing 3714 specimens derived from 100 malignancies and from 273 normal control tissues.Results:The expression of CAR was detected in all normal organs, except in the brain. Expression levels, however, displayed a broad range from being barely detectable (for example, in the thymus) to high abundance expression (for example, in the liver and gastric mucosa). In malignancies, a high degree of variability was notable also, ranging from significantly elevated CAR expression (for example, in early stages of malignant transformation and several tumours of the female reproductive system) to decreased CAR expression (for example, in colon and prostate cancer types).Conclusion:Our results provide a comprehensive insight into CAR expression in neoplasms and indicate that CAR may offer a valuable target for adenovirus-based therapy in a subset of carcinomas. Furthermore, these data suggest that CAR may contribute to carcinogenesis in an entity-dependent manner. PMID:24022195

Reeh, M; Bockhorn, M; Görgens, D; Vieth, M; Hoffmann, T; Simon, R; Izbicki, J R; Sauter, G; Schumacher, U; Anders, M



Species D Adenoviruses as Oncolytics Against B Cell Cancers  

PubMed Central

Purpose Oncolytic viruses are self-amplifying anti-cancer agents that make use of the natural ability of viruses to kill cells. Adenovirus serotype 5 (Ad5) has been extensively tested against solid cancers, but less so against B cell cancers since these cells do not generally express the coxsackie and adenoviral receptor (CAR). To determine if other adenoviruses might have better potency, we “mined” the adenovirus virome of 55 serotypes for viruses that could kill B cell cancers. Experimental Design 15 adenoviruses selected to represent Ad species B, C, D, E, and F were tested in vitro against cell lines and primary patient B cell cancers for their ability to infect, replicate in, and kill these cells. Select viruses were also tested against B cell cancer xenografts in immunodeficient mice. Results Species D adenoviruses mediated most robust killing against a range of B cell cancer cell lines, against primary patient marginal zone lymphoma cells, and against primary patient CD138+ myeloma cells in vitro. When injected into xenografts in vivo, single treatment with select species D viruses Ad26 and Ad45 delayed lymphoma growth. Conclusions Relatively unstudied species D adenoviruses have a unique ability to infect and replicate in B cell cancers as compared to other adenovirus species. These data suggest these viruses have unique biology in B cells and support translation of novel species D adenoviruses as oncolytics against B cell cancers.

Chen, Christopher Y.; Senac, Julien S.; Weaver, Eric A.; May, Shannon M.; Jelinek, Diane F.; Greipp, Philip; Witzig, Thomas; Barry, Michael A.



Cleavage of eukaryotic initiation factor eIF5B by enterovirus 3C proteases  

PubMed Central

The enteroviruses poliovirus (PV), Coxsackie B virus (CVB) and rhinovirus (HRV) are members of Picornaviridae that inhibit host cell translation early in infection. Enterovirus translation soon predominates in infected cells, but eventually also shuts off. This complex pattern of modulation of translation suggests regulation by a multifactorial mechanism. We report here that eIF5B is proteolytically cleaved during PV and CVB infection of cultured cells, beginning at 3 hours post-infection and increasing thereafter. Recombinant PV, CVB and HRV 3Cpro cleaved purified native rabbit eukaryotic initiation factor (eIF) 5B in vitro at a single site (VVEQ?G, equivalent to VMEQ?G479in human eIF5B) that is consistent with the cleavage specificity of enterovirus 3C proteases. Cleavage separates the N-terminal domain of eIF5B from its essential conserved central GTPase and C-terminal domains. 3Cpro-mediated cleavage of eIF5B may thus play an accessory role in the shut-off of translation that occurs in enterovirus-infected cells.

de Breyne, Sylvain; Bonderoff, Jennifer M.; Chumakov, Konstantin M.; Lloyd, Richard E.; Hellen, Christopher U. T.



Adenoviral vectors coated with PAMAM dendrimer conjugates allow CAR independent virus uptake and targeting to the EGF receptor.  


Adenovirus type 5 (Ad) is an efficient gene vector with high gene transduction potential, but its efficiency depends on its native cell receptors coxsackie- and adenovirus receptor (CAR) for cell attachment and ?(v)?(3/5) integrins for internalization. To enable transduction of CAR negative cancer cell lines, we have coated the negatively charged Ad by noncovalent charge interaction with cationic PAMAM (polyamidoamine) dendrimers. The specificity for tumor cell infection was increased by targeting the coated Ad to the epidermal growth factor receptor using the peptide ligand GE11, which was coupled to the PAMAM dendrimer via a 2 kDa PEG spacer. Particles were examined by measuring surface charge and size, the degree of coating was determined by transmission electron microscopy. The net positive charge of PAMAM coated Ad enhanced cellular binding and uptake leading to increased transduction efficiency, especially in low to medium CAR expressing cancer cell lines using enhanced green fluorescent protein or luciferase as transgene. While PAMAM coated Ad allowed for efficient internalization, coating with linear polyethylenimine induced excessive particle aggregation, elevated cellular toxicity and lowered transduction efficiency. PAMAM coating of Ad enabled successful transduction of cells in vitro even in the presence of neutralizing antibodies. Taken together, this study clearly proves noncovalent, charge-based coating of Ad vectors with ligand-equipped dendrimers as a viable strategy for efficient transduction of cells otherwise refractory to Ad infection. PMID:23281933

Vetter, Alexandra; Virdi, Kulpreet S; Espenlaub, Sigrid; Rödl, Wolfgang; Wagner, Ernst; Holm, Per S; Scheu, Christina; Kreppel, Florian; Spitzweg, Christine; Ogris, Manfred



Ultrasound-induced cavitation enhances the delivery and therapeutic efficacy of an oncolytic virus in an in vitro model.  


We investigated whether ultrasound-induced cavitation at 0.5 MHz could improve the extravasation and distribution of a potent breast cancer-selective oncolytic adenovirus, AdEHE2F-Luc, to tumour regions that are remote from blood vessels. We developed a novel tumour-mimicking model consisting of a gel matrix containing human breast cancer cells traversed by a fluid channel simulating a tumour blood vessel, through which the virus and microbubbles could be made to flow. Ultrasonic pressures were chosen to maximize either broadband emissions, associated with inertial cavitation, or ultraharmonic emissions, associated with stable cavitation, while varying duty cycle to keep the total acoustic energy delivered constant for comparison across exposures. None of the exposure conditions tested affected cell viability in the absence of the adenovirus. When AdEHE2F-Luc was delivered via the vessel, inertial cavitation increased transgene expression in tumour cells by up to 200 times. This increase was not observed in the absence of Coxsackie and Adenovirus Receptor cell expression, discounting sonoporation as the mechanism of action. In the presence of inertial cavitation, AdEHE2F-Luc distribution was greatly improved in the matrix surrounding the vessel, particularly in the direction of the ultrasound beam; this enabled AdEHE2F-Luc to kill up to 80% of cancer cells within the ultrasound focal volume in the gel 24 hours after delivery, compared to 0% in the absence of cavitation. PMID:21982902

Bazan-Peregrino, Miriam; Arvanitis, Costas D; Rifai, Bassel; Seymour, Leonard W; Coussios, Constantin-C



Splenomegaly in acute infections due to group A streptococci and viruses.  

PubMed Central

Over a period of 9 years in general practice temporary enlargement of the spleen was found in 29 episodes of pharyngitis or tonsillitis, in 2 episodes of acute upper respiratory tract infection other than pharyngitis and in 6 episodes of acute cervical lymphadenitis. In five patients more than one episode of illness associated with splenomegaly was recorded. In 26 of the 37 episodes a possible aetiology was identified. Evidence only of infection with group A streptococci was found in 14 episodes, adenoviruses or coxsackie B viruses were isolated alone in 4 episodes and in 4 episodes the only finding was the presence in the blood of more than occasional atypical mononuclear cells; in 4 episodes there was evidence of both streptococcal and viral infection. Episodes with evidence of streptococcal infection only tended to be of shorter duration and to be more evenly distributed over the year than were episodes without such evidence. Temporary splenomegaly was noted also in two children with varicella (one of whom also had streptococcal infection) and in an adult with probable rubella.

Higgins, P. M.



Human enterovirus 71 epidemics: what's next?  


Human enterovirus 71 (EV71) epidemics have affected various countries in the past 40 years. EV71 commonly causes hand, foot and mouth disease (HFMD) in children, but can result in neurological and cardiorespiratory complications in severe cases. Genotypic changes of EV71 have been observed in different places over time, with the emergence of novel genotypes or subgenotypes giving rise to serious outbreaks. Since the late 1990s, intra- and inter-typic recombination events in EV71 have been increasingly reported in the Asia-Pacific region. In particular, 'double-recombinant' EV71 strains belonging to a novel genotype D have been predominant in mainland China and Hong Kong over the last decade, though co-circulating with a minority of other EV71 subgenotypes and coxsackie A viruses. Continuous surveillance and genome studies are important to detect potential novel mutants or recombinants in the near future. Rapid and sensitive molecular detection of EV71 is of paramount importance in anticipating and combating EV71 outbreaks. PMID:24119538

Yip, Cyril C Y; Lau, Susanna K P; Woo, Patrick C Y; Yuen, Kwok-Yung



Paleobiogeography of the Onondaga Limestone in southeastern New York: A second shelf to basin ramp  

SciTech Connect

As a logical consequence of outcrop pattern and density, research on the Middle Devonian Onondaga Limestone has concentrated on the formation in an essentially east-west transect between the mid-Hudson Valley and Buffalo. Paleogeographic analysis reveals a symmetrical pattern, with the axis of the Appalachian Basin in central New York and the Edgecliff reefs flourishing in the eastern and western shelf regions. The subsurface reef trend of western New York and Pennsylvania is consistent with this model and is suggestive of a parallel reef trend in the east. However, recent examination of the formation in the southeastern part of the state has led to a pronounced reinterpretation of Onondaga paleobiogeography in that area. The authors have determined that there was a shallow carbonate shelf in the Helderberg-Coxsackie areas, a thick accumulation of shelf-margin bryozoan bafflestone between Leeds and Saugerties, and an even thicker accumulation of sparse to packed biocalcisiltites deposited on a carbonate ramp dipping southward into the Port Jervis area. It appears that this ramp occupied the northern margin of a structural base depocenter, located to the east of and not directly related to the topographic basin of central New York, which was centered in the Tristates vicinity from the Late Silurian until the early Middle Devonian.

Lindemann, R.H. (Skidmore Coll., Saratoga Springs, NY (United States). Dept. of Geology); Feldman, H.R. (Touro Coll., New York, NY (United States). Biology Dept.)



Clathrin adaptor AP1B controls adenovirus infectivity of epithelial cells  

PubMed Central

Adenoviruses invading the organism via normal digestive or respiratory routes require the Coxsackie-adenovirus receptor (CAR) to infect the epithelial barrier cells. Because CAR is a component of tight junctions and the basolateral membrane and is normally excluded from the apical membrane, most epithelia are resistant to adenoviruses. However, we discovered that a specialized epithelium, the retinal pigment epithelium (RPE), anomalously expressed CAR at the apical surface and was highly susceptible to adenovirus infection. These properties of RPE cells correlated with the absence of the epithelial-specific clathrin adaptor AP1B. Furthermore, knockdown of this basolateral sorting adaptor in adenovirus-resistant MDCK cells promoted apical localization of CAR and increased dramatically Adenovirus infectivity. Targeting assays showed that AP1B is required for accurate basolateral recycling of CAR after internalization. AP1B knock down MDCK cells missorted CAR from recycling endosomes to the apical surface. In summary, we have characterized the cellular machinery responsible for normal sorting of an adenovirus receptor and illustrated how tissue-specific variations in such machinery result in drastic changes in tissue-susceptibility to adenoviruses.

Diaz, Fernando; Gravotta, Diego; Deora, Ami; Schreiner, Ryan; Schoggins, John; Falck-Pedersen, Erik; Rodriguez-Boulan, Enrique



Update on molecular characterization of coxsackievirus B5 strains.  


Among Coxsackie B viruses, Coxsckievirus B5 is one of the most predominant serotypes in human, it is frequently associated with cases of neurological diseases, epidemics of meningitis and is a common cause of cardiomyopathy and diabetes. In the present study 27 isolates of Coxsackievirus B5 from North Africa, obtained from cerebrospinal fluid and stool samples of healthy individuals, patients with acute flaccid paralysis or aseptic meningitis were investigated by partial sequencing in the 5' half of the VP1 region and compared to the up-to-date published Coxsackievirus B5 sequences in the same genomic region. Four distinct genomic groups and ten different clusters were individualized. Most of the isolates from Algeria and Tunisia belonged to two clusters. For both, the sequences from North Africa clustered mainly with sequences from European countries, the majority isolated recently during the 2000s. The analysis of the alignment of amino-acids sequences in the VP1 gene revealed four major substitutions in strains from different clusters, we also noticed changes in the BC-loop region; this region is associated with viral antigenicity. This study permit to better identify circulating Coxsackievirus B5 strains throughout the world and their genetic relationship. The protein analysis showed changes that could imply some antigenic significance. J. Med. Virol. 83:1247-1254, 2011. © 2011 Wiley-Liss, Inc. PMID:21567427

Rezig, Dorra; Fares, Wasfi; Seghier, Mohamed; Yahia, Ahlem Ben; Touzi, Henda; Triki, Hinda



Cytokine and chemokine production by human pancreatic islets upon enterovirus infection.  


Enteroviruses of the human enterovirus B species (HEV-Bs) (e.g., coxsackie B viruses [CVBs] and echoviruses) have been implicated as environmental factors that trigger/accelerate type 1 diabetes, but the underlying mechanism remains elusive. The aim of this study was to gain insight into the cytokines and chemokines that are produced by human pancreatic islets upon infection with CVBs. To this end, we studied the response of human islets of Langerhans upon mock or CVB3 infection. Using quantitative PCR, we showed that upon CVB3 infection, transcription of interferon (IFN), IFN-stimulated genes, and inflammatory genes was induced. Analysis of secreted cytokines and chemokines by Luminex technology confirmed production and secretion of proinflammatory cytokines (e.g., interleukin [IL]-6 and tumor necrosis factor-?) as well as various chemotactic proteins, such as IFN-?-induced protein 10, macrophage inflammatory protein (MIP)-1?, MIP-1?, and IL-8. Infection with other HEV-Bs induced similar responses, yet their extent depended on replication efficiency. Ultra violet-inactivated CVB3 did not induce any response, suggesting that virus replication is a prerequisite for antiviral responses. Our data represent the first comprehensive overview of inflammatory mediators that are secreted by human islets of Langerhans upon CVB infection and may shed light on the role of enteroviruses in type 1 diabetes pathogenesis. PMID:22596052

Schulte, Barbara M; Lanke, Kjerstin H W; Piganelli, Jon D; Kers-Rebel, Esther D; Bottino, Rita; Trucco, Massimo; Huijbens, Richard J F; Radstake, Timothy R D J; Engelse, Marten A; de Koning, Eelco J P; Galama, Jochem M; Adema, Gosse J; van Kuppeveld, Frank J M



[Stomatitis in childhood, not always benign].  


Two boys of 1 and 16 year had painful buccal lesions and were admitted for dehydration. The younger had finger and toe blisters; the older, severely ill, had conjunctivitis, urethritis and skin lesions. Only symptomatic treatment with lidocaine gel and paracetamol gave good recovery. A 5-year-old Turkish girl had recurrent painful buccal ulcers which each time cleared up spontaneously. Stomatitis is common in childhood. Viral infections are the most common causes of stomatitis, in particular infections with herpes simplex virus (herpes gingivostomatitis), Coxsackie virus (herpangina, hand-foot-mouth-disease), chickenpox and infectious mononucleosis. Bacterial infections are rare and mostly secondary to the viral infections. In infants oral candidiasis (thrush) is a common cause of stomatitis. Most infections are self-limiting and reassurance of parents is important. Dehydration is a common complication and admission to hospital can be prevented by analgesics. The most important non-infectious conditions that cause stomatitis in children are recurrent aphthous stomatitis, erythema multiforme major (Stevens-Johnson syndrome), Behçet's disease, malignancy (leukaemia), immune-mediated disorders (agranulocytosis, cyclic neutropenia), traumata, blistering disorders of the skin and lichen planus. A complete history and a thorough physical examination usually give the correct diagnosis and further investigations are seldom necessary. PMID:11072515

Oudshoorn, A M; Ramaker, C



Functional and Selective Targeting of Adenovirus to High-Affinity Fc? Receptor I-Positive Cells by Using a Bispecific Hybrid Adapter  

PubMed Central

Adenovirus (Ad) efficiently delivers its DNA genome into a variety of cells and tissues, provided that these cells express appropriate receptors, including the coxsackie-adenovirus receptor (CAR), which binds to the terminal knob domain of the viral capsid protein fiber. To render CAR-negative cells susceptible to Ad infection, we have produced a bispecific hybrid adapter protein consisting of the amino-terminal extracellular domain of the human CAR protein (CARex) and the Fc region of the human immunoglobulin G1 protein, comprising the hinge and the CH2 and CH3 regions. CARex-Fc was purified from COS7 cell supernatants and mixed with Ad particles, thus blocking Ad infection of CAR-positive but Fc receptor-negative cells. The functionality of the CARex domain was further confirmed by successful immunization of mice with CARex-Fc followed by selection of a monoclonal anti-human CAR antibody (E1-1), which blocked Ad infection of CAR-positive cells. When mixed with Ad expressing eGFP, CARex-Fc mediated an up to 250-fold increase of transgene expression in CAR-negative human monocytic cell lines expressing the high-affinity Fc? receptor I (CD64) but not in cells expressing the low-affinity Fc? receptor II (CD32) or III (CD16). These results open new perspectives for Ad-mediated cancer cell vaccination, including the treatment of acute myeloid leukemia.

Ebbinghaus, Christina; Al-Jaibaji, Ahmed; Operschall, Elisabeth; Schoffel, Angelika; Peter, Isabelle; Greber, Urs F.; Hemmi, Silvio



Screening of some Tanzanian medicinal plants from Bunda district for antibacterial, antifungal and antiviral activities.  


Extracts from 50 plant parts obtained from 39 different plants belonging to 22 families used to treat infectious diseases in Bunda district, Tanzania, were screened against twelve microorganisms, including the bacteria Bacillus cereus, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Salmonella typhimurium, the fungi Aspergillus niger, Candida albicans, and the viruses Herpes Simplex Virus type 1, Vesicular Stomatitis Virus T2, Coxsackie B2 and Semliki Forest A7. The highest activity was obtained for the n-hexane extract of Elaeodendron schlechteranum root bark against the Gram-positive bacteria Bacillus cereus (MIC 0.97 microg/ml and MBC 1.95 microg/ml) and Staphylococcus aureus (MIC 3.90 microg/ml and MBC 31.25 microg/ml). Gram-negative bacteria were less sensitive. Only Balanites aegyptiaca stem bark exhibited a high antifungal activity against Candida albicans (MIC 125 microg/ml and MFC 250 microg/ml). Extracts from four plants; Lannea schweinfurthii, Combretum adenogonium, Ficus sycomorus and Terminalia mollis showed strong antiviral activity with RF values of 10(3) and 10(4) against Herpes Simplex Virus type 1 at various concentrations. Our results support, at least in part, the use of most plants as claimed by traditional healers/informants especially against the Gram-positive bacteria Bacillus cereus and Staphylococcus aureus. PMID:18582554

Maregesi, Sheila Mgole; Pieters, Luc; Ngassapa, Olipa David; Apers, Sandra; Vingerhoets, Rita; Cos, Paul; Berghe, Dirk A Vanden; Vlietinck, Arnold J



A-type proanthocyanidins from lychee seeds and their antioxidant and antiviral activities.  


Two new A-type trimeric proanthocyanidins with two doubly bonded interflavanoid linkages, litchitannin A1 [epicatechin-(2??O?7,4??6)-epicatechin-(2??O?7,4??8)-catechin] (1) and litchitannin A2 [epicatechin-(2??O?7,4??6)-epicatechin-(2??O?7,4??6)-epicatechin] (2), were isolated from lychee (Litchi chinensis Sonn. cv. Heiye) seeds together with aesculitannin A (3), epicatechin-(2??O?7,4??8)-epiafzelechin-(4??8)-epicatechin (4), proanthocyanidin A1 (5), proanthocyanidin A2 (6), proanthocyanidin A6 (7), epicatechin-(7,8-bc)-4?-(4-hydroxyphenyl)-dihydro-2(3H)-pyranone (8), and epicatechin (9). Their structures were elucidated on the basis of spectroscopic and chemical evidence. It is the first time that compounds 1-4, 7, and 8 have been reported in this species. Compounds 1-9 showed more potent antioxidant activity than L-ascorbic acid with ferric reducing antioxidant power (FRAP) values of 3.71-24.18 mmol/g and IC50 values of 5.25-20.07 ?M toward DPPH radicals. Moreover, litchitannin A2 (2) was found to exhibit in vitro antiviral activity against coxsackie virus B3 (CVB3) and compounds 3 and 6 displayed antiherpes simplex virus 1 (HSV-1) activity. PMID:20964424

Xu, Xinya; Xie, Haihui; Wang, Yifei; Wei, Xiaoyi



GRP78: a multifunctional receptor on the cell surface.  


The 78 kDa glucose-regulated protein (GRP78) is an endoplasmic reticulum chaperone, whose function is generally thought to be restricted to controlling the structural maturation of nascent glycoproteins. However, GRP78 also is expressed on the cell surface where it functions as a receptor for a wide variety of ligands, behaving as an autoantigen for several classes of autoantibodies. GRP78 is a signaling receptor for activated alpha2-macroglobulin, plasminogen kringle 5, and microplasminogen, and it plays a critical role in viral entry of coxsackie B, and dengue fever viruses. GRP78 is also implicated in the regulation of tissue factor procoagulant activity and functions as a receptor for angiogenic peptides via a mechanism independent of the VEGF receptor. Cell surface GRP78 is found associated with such diverse proteins as the voltage-dependent anion channel (VDAC), the major histocompatibility complex class I (MHC-I), the teratocarcinoma-derived growth factor I (Cripto), and the DnaJ-like protein MTJ-1. These associations suggest a unique GRP78 cell surface topography, which appears to be compartmentalized to respond differently to agonists that bind to its N- or C-terminal domains. Here, we discuss the significance of these associations, and the possible mechanisms involved in the transportation of GRP78 from the cytosol to the cell surface. PMID:19331544

Gonzalez-Gronow, Mario; Selim, Maria Angelica; Papalas, John; Pizzo, Salvatore V



[New findings in type I diabetes].  


The author present a review of contemporary views on the pathogenesis of type I diabetes, in particular with regard to recent research of European and non-European diabetelogical departments. In the aetiopathogenesis attention is drawn to genetic influences, external factor and stimuli, the importance of some virus groups (Picornidae, Mengo 2T virus, Coxsackie B5, cytomegaloviruses, congenital rubeola syndrome etc.). Special attention is paid to the autoimmune theory of type I diabetes, the importance of different types of anti- beta-cell antibodies, insulin antibodies, antibodies against protein 64-KD of the islets, pro-insulin antibody PAA, and other stimuli and interferences (oxidation stress, non-enzymatic glycosylation etc.). As to treatment, the author mentions immunosuppressive treatment (cyclosporin A, azathioprine, prednisone) and transplantation of the pancreas. The author presents his own experience with a group of 2040 diabetics; in a group of 253 juvenile diabetics he followed-up the development of micro- and macroangiopathic complications in the course of 25 years. He proved an increase of so-called late diabetic complications in particular vascular ones along with the lengthening period of diabetes, with advancing age during its clinical manifestation. In the conclusion the author draws attention to the importance of multi-centre studies and new laboratory techniques. PMID:8131177

Syllaba, J



Homomultimerization of the Coxsackievirus 2B Protein in Living Cells Visualized by Fluorescence Resonance Energy Transfer Microscopy  

PubMed Central

The 2B protein of enteroviruses is the viral membrane-active protein that is responsible for the modifications in host cell membrane permeability that take place in enterovirus-infected cells. The 2B protein shows structural similarities to the group of lytic polypeptides, polypeptides that permeate membranes either by forming multimeric membrane-integral pores or, alternatively, by lying parallel to the lipid bilayer and disturbing the curvature and symmetry of the membrane. Our aim is to gain more insight into the molecular architecture of the 2B protein in vivo. In this study, the possible existence of multimers of the coxsackie B3 virus 2B protein in single living cells was explored by fluorescence resonance energy transfer (FRET) microscopy. FRET between fusion proteins 2B-ECFP and 2B-EYFP (enhanced cyan and yellow fluorescent variants of green fluorescent protein) was monitored by using spectral imaging microscopy (SPIM) and fluorescence lifetime imaging microscopy (FLIM). Both techniques revealed the occurrence of intermolecular FRET between 2B-ECFP and 2B-EYFP, providing evidence for the formation of protein 2B homomultimers. Putative models for the mode of action of the membrane-active 2B protein and the formation of membrane-integral pores by 2B multimers are discussed.

van Kuppeveld, Frank J. M.; Melchers, Willem J. G.; Willems, Peter H. G. M.; Gadella, Jr., Theodorus W. J.



The expression and significance of proto-oncogene c-fos in viral myocarditis  

PubMed Central

Background c-fos may play a role in the pathogenesis of some diseases. The expression and function of c-fos in viral myocarditis (VMC) have not yet been reported. To study the change and significance of proto-oncogene c-fos in VMC is the objective of this experiment. Methods An animal model of VMC was established via coxsackie virus B3 inoculation. VMC mice were then treated with a c-fos monoclonal antibody and isoproterenol and the protein and mRNA expression of c-fos were studied via immunohistochemical analysis and in situ hybridization. Results were simultaneously analyzed for the significance of c-fos expression in mice with VMC. Results Myocardial necrosis and cell infiltration decreased after treatment with c-fos monoclonal antibody compared to control mice, while myocardial necrosis and cell infiltration were increased after treatment with isoproterenol. Positive cardiomyocytes with c-Fos expression increased at 3, 5, 7, 9, and 15 days after virus inoculation in VMC mice compared to control mice, while returning to almost normal levels at 35 days. The expression level of c-fos mRNA at 3 and 7 days after virus inoculation in VMC mice was also higher than that of control mice. Conclusions c-fos expression in the cardiomyocytes of VMC mice is significantly increased, c-fos plays an important role in myocardial lesions. The apparent increase in expression of c-fos is likely to be involved in the pathogenesis of VMC.



[Possible factors associated with epidemic neuropathy in Cuba].  


Of the 115 patients who suffered from epidemic neuropathy and received attention from March 16 de April 30, 1993, at the Center for Medical and Surgical Research (CMSR), in Havana City, 114 controls were selected at random in the community. In the univariate analysis it was found a significant association mainly with the smoking habit (odds ratio = 3.45 [95% confidence interval 1.85 to 6.35)] and with the consumption of edible fat of uncertified procedence (odds ratio = 2.77 [95% confidence interval 1.34 to 5.8]). All were ratified in the multivariate logistical regression analysis. Association (odds ratio = 8.8 [95% confidence interval 2.58 to 30.55] with the presence of antibodies against the Coxsackie virus A-9 strain 47 was also found among the 182 individuals who had serological test. It seems that the toxic factor derived from the smoking habit, together with the deficiency of some nutrients due to the changes occurred in the diet, as well as the contact with the isolated virus in one of the patients facilitated the clinical expression of this disease and therefore in the origin of the epidemics. PMID:9842269

Pérez Rodríguez, A; Isla García, A; Fernández, I; Más Lago, P; García Rodríguez, A I; Rodríguez Concepción, A



Viral myocarditis and dilated cardiomyopathy: mechanisms, manifestations, and management  

PubMed Central

Viral infection of the heart is relatively common and usually of little consequence. It can, however, lead to substantial cardiac damage and severe acute heart failure. It can also evolve into the progressive syndrome of chronic heart failure. Recent studies have gone some way towards unravelling the complex mechanisms underlying the heart muscle damage that occurs after viral infection. These studies have lent support to both immune and viral mediated (independent of an immune response) cardiac damage. Acute myocarditis can present in various ways, and it may be a cause of sudden death in an otherwise healthy young adult. New treatments for viral heart disease are awaited. In the meanwhile, the haemodynamic support of patients with acute left ventricular failure caused by myocarditis should be aggressive, to allow for the possibility of spontaneous recovery. Contemporary trials of treatment in chronic heart failure secondary to dilated cardiomyopathy support the use of angiotensin converting enzyme inhibitors, ? adrenoceptor blockers, and spironolactone in such patients.???Keywords: myocarditis; heart failure; coxsackie B virus; dilated cardiomyopathy

Kearney, M; Cotton, J; Richardson, P; Shah, A



Enhanced Prostate Cancer Gene Transfer and Therapy Using a Novel Serotype Chimera Cancer Terminator Virus (Ad.5/3-CTV).  


Few options are available for treating patients with advanced prostate cancer (PC). As PC is a slow growing disease and accessible by ultrasound, gene therapy could provide a viable option for this neoplasm. Conditionally replication-competent adenoviruses (CRCAs) represent potentially useful reagents for treating PC. We previously constructed a CRCA, cancer terminator virus (CTV), which showed efficacy both in vitro and in vivo for PC. The CTV was generated on a serotype 5-background (Ad.5-CTV) with infectivity depending on Coxsackie-Adenovirus Receptors (CARs). CARs are frequently reduced in many tumor types, including PCs thereby limiting effective Ad-mediated therapy. Using serotype chimerism, a novel CTV (Ad.5/3-CTV) was created by replacing the Ad.5 fiber knob with the Ad.3 fiber knob thereby facilitating infection in a CAR-independent manner. We evaluated Ad.5/3-CTV in comparison with Ad.5-CTV in low CAR human PC cells, demonstrating higher efficiency in inhibiting cell viability in vitro. Moreover, Ad.5/3-CTV potently suppressed in vivo tumor growth in a nude mouse xenograft model and in a spontaneously induced PC that develops in Hi-myc transgenic mice. Considering the significant responses in a Phase I clinical trial of a non-replicating Ad.5-mda-7 in advanced cancers, Ad.5/3-CTV may exert improved therapeutic benefit in a clinical setting. J. Cell. Physiol. 229: 34-43, 2014. © 2013 Wiley Periodicals, Inc. PMID:23868767

Azab, Belal M; Dash, Rupesh; Das, Swadesh K; Bhutia, Sujit K; Sarkar, Siddik; Shen, Xue-Ning; Quinn, Bridget A; Dent, Paul; Dmitriev, Igor P; Wang, Xiang-Yang; Curiel, David T; Pellecchia, Maurizio; Reed, John C; Sarkar, Devanand; Fisher, Paul B



Unexpected hazard of illegal immigration: Outbreak of viral myocarditis exacerbated by confinement and deprivation in a shipboard cargo container.  


We present a group of 18 illegal immigrant stowaways who arrived in a shipboard cargo container suffering from gastroenteritis, dehydration, and malnutrition and showing evidence of viral myocarditis in 3 of 4 fatalities. Our investigation included an evaluation of the 2-week ocean voyage, analysis of medical records and laboratory results of the survivors, autopsies on the decedents, and viral studies on their heart tissue. Of 3 stowaways who died shipboard, 2 showed lymphocytic myocarditis and 1 could not be evaluated histologically due to decomposition. A fourth stowaway died 4 months after arrival with dilated cardiomyopathy and lymphocytic myocarditis. Reverse-transcriptase polymerase chain reaction and nucleotide sequencing of viral isolates from the decedents' heart tissues demonstrated Coxsackie virus B3 genome. We believe that these cases represent an outbreak of viral myocarditis, exacerbated by acute dehydration and malnutrition, due to confinement within the shipping container. Our evidence indicates that close confinement promoted the spread of the virus, and nutritional deprivation increased the stowaways' vulnerability. Furthermore, our observations support the conclusion, based on experimental studies, that nutritionally induced oxidative stress increased the virulence of the etiologic viral agent. In summary, these cases represent a potential infectious disease hazard of illegal immigration. PMID:15166761

Li, Melissa K; Beck, Melinda A; Shi, Qing; Harruff, Richard C



Adenovirus triggers macropinocytosis and endosomal leakage together with its clathrin-mediated uptake  

PubMed Central

Adenovirus type 2 (Ad2) binds the coxsackie B virus Ad receptor and is endocytosed upon activation of the ?v integrin coreceptors. Here, we demonstrate that expression of dominant negative clathrin hub, eps15, or K44A-dynamin (dyn) inhibited Ad2 uptake into epithelial cells, indicating clathrin-dependent viral endocytosis. Surprisingly, Ad strongly stimulated the endocytic uptake of fluid phase tracers, coincident with virus internalization but without affecting receptor-mediated transferrin uptake. A large amount of the stimulated endocytic activity was macropinocytosis. Macropinocytosis depended on ?v integrins, PKC, F-actin, and the amiloride-sensitive Na+/H+ exchanger, which are all required for Ad escape from endosomes and infection. Macropinocytosis stimulation was not a consequence of viral escape, since it occurred in K44A-dyn–expressing cells. Surprisingly, 30–50% of the endosomal contents were released into the cytosol of control and also K44A-dyn–expressing cells, and the number of fluid phase–positive endosomes dropped below the levels of noninfected cells, indicating macropinosomal lysis. The release of macropinosomal contents was Ad dose dependent, but the presence of Ad particles on macropinosomal membranes was not sufficient for contents release. We conclude that Ad signaling from the cell surface controls the induction of macropinosome formation and leakage, and this correlates with viral exit to the cytosol and infection.

Meier, Oliver; Boucke, Karin; Hammer, Silvija Vig; Keller, Stephan; Stidwill, Robert P.; Hemmi, Silvio; Greber, Urs F.



Two different PCR assays to detect enteroviral RNA in CSF samples from patients with acute aseptic meningitis.  


Two polymerase chain reaction (RT-PCR) assays were developed to allow rapid detection of enteroviral RNA in cerebrospinal fluid samples (CSF). Primers homologous to the conserved 5' noncoding region of the enterovirus genome were designed. The RT-PCR product size was approximately 500 bp (479 bp for Poliovirus, 500 bp for Coxsackievirus) and was visualized using ethidium bromide-stained gels. Assay 1 utilized Moloney Murine Leukaemia Virus Reverse Transcriptase (MMLV-RTase) for reverse transcription and Taq polymerase for subsequent PCR. Assay 2 utilized a thermoactive DNA polymerase of Thermus thermophilus (rTth enzyme) for both reverse transcription and DNA amplification. In addition, in Assay 2 reverse transcription and PCR were accomplished within the same reaction tube. Both assays detected between 1 and 0.02 TCID50 of prototype strains of Polio and Coxsackie type B viruses propagated in VERO cell and spiked in a pooled preparation of CSF samples from patients with noninfective neurological disorders. However, Assay 1 was 10-fold more sensitive than Assay 2 when applied to the detection of enteroviral RNA in CSF samples from patients with etiologically well characterized acute aseptic meningitis. PMID:8636706

Casas, I; Klapper, P E; Cleator, G M; Echevarría, J E; Tenorio, A; Echevarría, J M



Microbiology and management of neonatal necrotizing enterocolitis.  


Necrotizing enterocolitis (NEC) is a clinical syndrome of ischemic necrosis of the bowel of multiple etiological factors that include the presence of intestinal ischemia, abnormal bacterial flora, and intestinal mucosal immaturity. Numerous reports have implied that the fecal microflora may contribute to the pathogenesis of NEC. A broad range of organisms generally found in the distal gastrointestinal tract have been recovered from the peritoneal cavity and blood of infants with NEC. The predominant organisms include Enterobacteriaceae (i.e., Escherichia coli, Klebsiella pneumoniae) , Clostridium spp., enteric pathogens (salmonellae, Coxsackie B2 virus, coronavirus, rotavirus), and potential pathogens (Bacteroides fragilis). The goals of the initial management is preventing ongoing damage, restoring hemostasis, and minimizing complications. Medical management includes withholding oral feeding, placement of nasogastric tube, abdominal decompression, paracentesis, vigorous intravenous hydration containing electrolytes and calories, support of the circulation, administration of antibiotics, and surveillance for deterioration or complications that require surgical intervention. Indications for surgery include clinical deterioration, perforation, peritonitis, obstruction, and abdominal mass. Prevention remains crucial to decrease the incidence of NEC. Preventive methods include cautious feeding regimens, the use of maternal breast milk, and the use of probiotics. PMID:18236362

Brook, Itzhak



Infectious triggers in type 1 diabetes: is there a case for epitope mimicry?  


Environmental factors are the main contributors to type 1 diabetes (T1D) pathogenesis, yet they remain unidentified. Enteroviruses are proposed candidate triggers due to temporal correlations between infection and T1D autoimmunity and to detection of viral proteins in diseased islets. However, such correlations are not universal and may be relatively uncommon. Furthermore, evidence of a cause-effect relationship is lacking, as infection of non-obese diabetic mice with Coxsackie enteroviruses can either trigger or blunt disease. The proposed mechanisms are either non-antigen-specific (i.e. ?-cell destruction and release of sequestered antigens, islet inflammation) or antigen-specific (i.e. epitope mimicry, by which immune responses to enteroviruses may be diverted against homologous ?-cell antigens). The case for the latter mechanisms is even less stringent, as there is little evidence of promiscuous antigen recognition at the single T-cell level. Other infectious agents may thus be implicated. Demonstration of their role will require fulfilling the Koch's postulates, namely isolation of the agent preferentially in T1D patients, including before disease onset; and T1D induction when the agent is inoculated into mice. The same is needed for cross-reactive T cells to support epitope mimicry mechanisms. Generation of alternative (humanized) mouse models that could be challenged with candidate microbes is needed. PMID:24003924

Afonso, G; Mallone, R



Calcium Gluconate in Phosphate Buffered Saline Increases Gene Delivery with Adenovirus Type 5  

PubMed Central

Background Adenoviruses are attractive vectors for gene therapy because of their stability in vivo and the possibility of production at high titers. Despite exciting preclinical data with various approaches, there are only a few examples of clear efficacy in clinical trials. Effective gene delivery to target cells remains the key variable determining efficacy and thus enhanced transduction methods are important. Methods/Results We found that heated serum could enhance adenovirus 5 mediated gene delivery up to twentyfold. A new protein-level interaction was found between fiber knob and serum transthyretin, but this was not responsible for the observed effect. Instead, we found that heating caused the calcium and phosphate present in the serum mix to precipitate, and this was responsible for enhanced gene delivery. This finding could have relevance for designing preclinical experiments with adenoviruses, since calcium and phosphate are present in many solutions. To translate this into an approach potentially testable in patients, we used calcium gluconate in phosphate buffered saline, both of which are clinically approved, to increase adenoviral gene transfer up to 300-fold in vitro. Gene transfer was increased with or without heating and in a manner independent from the coxsackie-adenovirus receptor. In vivo, in mouse studies, gene delivery was increased 2-, 110-, 12- and 13-fold to tumors, lungs, heart and liver and did not result in increased pro-inflammatory cytokine induction. Antitumor efficacy of a replication competent virus was also increased significantly. Conclusion In summary, adenoviral gene transfer and antitumor efficacy can be enhanced by calcium gluconate in phosphate buffered saline.

Ahonen, Marko T.; Diaconu, Iulia; Pesonen, Sari; Kanerva, Anna; Baumann, Marc; Parviainen, Suvi T.; Spiller, Brad



CAR mediates efficient tumor engraftment of mesenchymal type lung cancer cells.  


The coxsackie-adenovirus receptor (CAR) is a developmentally regulated intercellular adhesion molecule that was previously observed to be required for efficient tumor formation. To confirm that observation, we compared the tumorigenicity of clonally derived test and control cell subsets that were genetically modified for CAR. Silencing CAR in lung cancer cells with high constitutive expression reduced engraftment efficiency. Conversely, overexpressing CAR in lung cancer cells with low constitutive expression did not affect tumor formation or growth kinetics. A blocking antibody to the extracellular domain of CAR inhibited tumor engraftment, implicating that domain as being important to this process. However, differences in adhesion properties attributable to this domain (barrier function and aggregation) could not be distinguished in the test groups in vitro, and the mechanisms underlying CAR's contribution to tumor engraftment remain elusive. Because high CAR cells displayed a spindle-shaped morphology at baseline, we considered whether this expression was an accompaniment of other mesenchymal features in these lung cancer cells. Molecular correlates of CAR were compared in model epithelial and mesenchymal type lung cancer cells. CAR expression is associated with an absence of E-cadherin, diminished expression of alpha- and gamma-catenin, and increased Zeb1, Snail, and vimentin expression in lung cancer cells. In contrast, epithelial type (NCI-H292, Calu3) lung cancer cells show comparatively low CAR expression. These data suggest that if the mesenchymal cell phenotype is an accurate measure of an undifferentiated and invasive state, then CAR expression may be more closely aligned with this phenotype of lung cancer cells. PMID:19506548

Veena, Mysore S; Qin, Min; Andersson, Asa; Sharma, Sherven; Batra, Raj K



Crystal structure of Swine vesicular disease virus and implications for host adaptation.  


Swine vesicular disease virus (SVDV) is an Enterovirus of the family Picornaviridae that causes symptoms indistinguishable from those of foot-and-mouth disease virus. Phylogenetic studies suggest that it is a recently evolved genetic sublineage of the important human pathogen coxsackievirus B5 (CBV5), and in agreement with this, it has been shown to utilize the coxsackie and adenovirus receptor (CAR) for cell entry. The 3.0-A crystal structure of strain UK/27/72 SVDV (highly virulent) reveals the expected similarity in core structure to those of other picornaviruses, showing most similarity to the closest available structure to CBV5, that of coxsackievirus B3 (CBV3). Features that help to cement together and rigidify the protein subunits are extended in this virus, perhaps explaining its extreme tolerance of environmental factors. Using the large number of capsid sequences available for both SVDV and CBV5, we have mapped the amino acid substitutions that may have occurred during the supposed adaptation of SVDV to a new host onto the structure of SVDV and a model of the SVDV/CAR complex generated by reference to the cryo-electron microscopy-visualized complex of CBV3 and CAR. The changes fall into three clusters as follows: one lines the fivefold pore, a second maps to the CAR-binding site and partially overlaps the site for decay accelerating factor (DAF) to bind to echovirus 7 (ECHO7), and the third lies close to the fivefold axis, where the low-density lipoprotein receptor binds to the minor group of rhinoviruses. Later changes in SVDV (post-1971) map to the first two clusters and may, by optimizing recognition of a pig CAR and/or DAF homologue, have improved the adaptation of the virus to pigs. PMID:12692248

Fry, Elizabeth E; Knowles, Nick J; Newman, John W I; Wilsden, Ginette; Rao, Zihe; King, Andrew M Q; Stuart, David I



Innate immunity in early chordates and the appearance of adaptive immunity.  


In the urochordate Ciona intestinalis some membrane Immunoglobulin superfamily members with ancestral features of antigen receptors are homologs of vertebrate adhesion molecules acting as virus receptors. They include the following: the junction adhesion molecule (reovirus receptor) (JAM), the Cortical thymocyte marker of Xenopus (CTX family) (Coxsackie's virus receptor) and the poliovirus receptor (PVR). In humans these genes belong to the same linkage group, of which 4 paralogous groups exist. This situation is consistent with the notion that the Ciona set of genes would correspond to a preduplication state. In addition, the human region 3q13 and its paralogs, harbour genes remotely related to the nectin family that can be detected in Protostomes (human CRTAM and CD80-86 related to Drosophila Beat). In addition, this linkage group contains several CDs important for the immune system CD166, CD47 and many members of the tetraspanin family. The VC1-like core of the nectin is homologous to the VCI core of the MHC-linked tapasin and to the VC1 segments of, for example, specific antigen receptors of vertebrates, and could be related to a primitive antigen receptor gene. It is suggested that the virus binding property of the members of this family was exploited, and that they were recruited in the vertebrate immune system following the introduction of the somatic rearrangement machinery. In this way the adaptive immune system could have developed from a set of receptors involved in a primitive local innate immunity involving NF-kappaB-mediated apoptosis. PMID:15330258

Du Pasquier, Louis



An epidemiological approach to the understanding and control of acute respiratory infections in Indian children.  


Epidemiology is emerging as a promising tool for understanding and interpreting diseases in all dimensions and identifying levels of intervention for their control or eradication. Coupled with the discipline of health administration and management, epidemiology can offer viable and pragmatic solutions for tackling disease problems. Eradication of small pox in the recent past and the formulation of a strategy for the control of the modern epidemic of AIDS are 2 best known examples. An epidemiological approach to acute respiratory tract infection (ARI) in children can improve the understanding of the disease and help to prepare the ground for effective control. The principal areas of epidemiology are natural history, intervention strategy, and epidemiological inquiry. The principal viruses that have been identified to lead to ARI infection on the basis of systematic studies are rhinoviruses (100 different serotypes), parainfluenza viruses (several serotypes), respiratory syncytial virus, adenoviruses (about 8 important serotypes in children), enteroviruses (ECHO and Coxsackie with more than 70 serotypes), herpes simplex virus, measles virus, and Epstein-Barr virus. Based on the natural history of ARI, the scope of application of the 5 levels of intervention may be discussed individually as health promotion, specific protection, early diagnosis and prompt treatment, disability limitation, rehabilitation, and epidemiological inquiry. Epidemiological techniques may contribute substantially to the understanding and management of ARI control by: a) defining the ARI problem and its magnitude and behavior in relation to time, place, and person; b) identifying the epidemiological correlates of ARI in terms of agent, host, and environment, and their impact on morbidity and mortality in children; c) introducing epidemiological surveillance and monitoring techniques for effective supervision of intervention activities; and d) conducting longitudinal observational studies to evaluate efficiency and effectiveness of various intervention alternatives for ARI control. PMID:12318658

Dhar, G M


Low-dose etoposide enhances telomerase-dependent adenovirus-mediated cytosine deaminase gene therapy through augmentation of adenoviral infection and transgene expression in a syngeneic bladder tumor model.  


The human telomerase reverse transcriptase (hTERT) promoter can selectively drive transgene expression in many telomerase-positive human cancer cells. Here we evaluated combination therapy of adenoviral vector Ad-hTERT-CD encoding E. coli cytosine deaminase (CD) driven by the hTERT promoter and low-dose etoposide (0.1 microg/mL) for treating bladder cancer. Ad-hTERT-CD conferred sensitivity to 5-fluorocytosine (5-FC) in bladder cancer cells, which could be enhanced by etoposide treatment, but not in normal cells. Such effect was correlated with up-regulation of hypoxia-inducible factor (HIF)-1alpha expression. By contrast, etoposide activated p53 and down-regulated hTERT promoter activity in normal cells. Etoposide also increased adenoviral infection via enhancement of coxsackie-adenovirus receptor expression on bladder cancer and normal cells. Combination index analysis revealed that combined therapy of Ad-hTERT-CD (10(9) plaque-forming units)/5-FC (200 mg/kg) with etoposide (2 mg/kg) synergistically suppressed tumor growth and prolonged survival in mice bearing syngeneic MBT-2 bladder tumors. This combination therapy regimen induced complete tumor regression and generated antitumor immunity in 75% of tumor-bearing mice. Furthermore, increased infiltrating CD4(+) and CD8(+) T cells and necrosis within tumors were found in mice receiving combination therapy of Ad-hTERT-CD and etoposide compared with those treated with either treatment alone. Thus, the potential high therapeutic index of the combination therapy may be an appealing therapeutic intervention for bladder cancer. Furthermore, because a majority of human tumors exhibit high telomerase activity, adenovirus-mediated CD gene therapy driven by the hTERT promoter in combination with low-dose etoposide may be applicable to a broad spectrum of cancers. PMID:17047058

Shieh, Gia-Shing; Shiau, Ai-Li; Yo, Yi-Te; Lin, Pey-Ru; Chang, Chao-Ching; Tzai, Tzong-Shin; Wu, Chao-Liang



In vitro culture of embryonic mouse intestinal epithelium: cell differentiation and introduction of reporter genes  

PubMed Central

Background Study of the normal development of the intestinal epithelium has been hampered by a lack of suitable model systems, in particular ones that enable the introduction of exogenous genes. Production of such a system would advance our understanding of normal epithelial development and help to shed light on the pathogenesis of intestinal neoplasia. The criteria for a reliable culture system include the ability to perform real time observations and manipulations in vitro, the preparation of wholemounts for immunostaining and the potential for introducing genes. Results The new culture system involves growing mouse embryo intestinal explants on fibronectin-coated coverslips in basal Eagle's medium+20% fetal bovine serum. Initially the cultures maintain expression of the intestinal transcription factor Cdx2 together with columnar epithelial (cytokeratin 8) and mesenchymal (smooth muscle actin) markers. Over a few days of culture, differentiation markers appear characteristic of absorptive epithelium (sucrase-isomaltase), goblet cells (Periodic Acid Schiff positive), enteroendocrine cells (chromogranin A) and Paneth cells (lysozyme). Three different approaches were tested to express genes in the developing cultures: transfection, electroporation and adenoviral infection. All could introduce genes into the mesenchyme, but only to a small extent into the epithelium. However the efficiency of adenovirus infection can be greatly improved by a limited enzyme digestion, which makes accessible the lateral faces of cells bearing the Coxsackie and Adenovirus Receptor. This enables reliable delivery of genes into epithelial cells. Conclusion We describe a new in vitro culture system for the small intestine of the mouse embryo that recapitulates its normal development. The system both provides a model for studying normal development of the intestinal epithelium and also allows for the manipulation of gene expression. The explants can be cultured for up to two weeks, they form the full repertoire of intestinal epithelial cell types (enterocytes, goblet cells, Paneth cells and enteroendocrine cells) and the method for gene introduction into the epithelium is efficient and reliable.

Quinlan, Jonathan M; Yu, Wei-Yuan; Hornsey, Mark A; Tosh, David; Slack, Jonathan MW



[Chemical consitituents from root of Isatis indigotica].  


Thirty-three compounds were isolated from the root decoction of Isatis indigotica by using a combination of various chromatographic techniques including silica gel, macroporous adsorbent resin, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by spectroscopic data as (+)-dehydrovomifoliol (1), (S)-(+)-abscisic acid (2), vomifoliol (3), cyclo (L-Phe-L-Leu) (4), cyclo(L-Phe-L-Tyr) (5), cyclo(L-Tyr-L-Leu) (6), cyclo(L-Pro-L-Tyr) (7), evofolin B (8), (+)-syringaresinol (9), (-)-(7R,7'R,8S,8'S)-4,4'-dihydroxy-3-methoxy-7,9';7',9-diepoxy-lignan (10), (-)-medioresinol (11), (+) -(7R,7'R,8S,8'S) -neo-olivil (12), (-) -5-methoxyisolariciresinol (13), 1,3-dihydro-2H-indol-2-one (14), isalexin (15), dihydroneoascorbigen (16), indican (17), (-) -(S) -cyanomethyl-3-hydroxyoxindole (18), isoformononetein (19), calycosin (20), stigamast-5-ene-3beta-ol-7-one (21), acetovanillone (22), 3, 5-dimethoxy-4-hydroxyacetophenone (23), dihydroconiferyl alcohol (24), dihyroferulic acid (25), 3-hydroxy-1-(4-hydroxyphenyl) propan-1-one (26), beta-hydroxypropiovanillone (27), 4-aminobenzoic acid (28), 3-(4-hydroxyphenyl) propan-1-ol (29), 4-(2-hydroxyethyl) phenol (30), 2-methoxy-4-vinylphenol (31), pyrocatechol (32), and 4-pentenamide (33). These compounds were isolated from the root of I. indigotica for the first time. In preliminary in vitro assays, compound 19 showed activity against the influenza virus A/Hanfang/359/95 (H3N2), the herpes simplex virus 1 (HSV-1), and Coxsackie virus B3 (Cox-B3), with IC50 values of 2.06, 6.84, and 8.70 micromol x L(-1), respectively, but other compounds were in-active at a concentration of 1.0 x 10 x (-5) mol x L(-1). PMID:23944031

Wang, Xiao-Liang; Chen, Ming-Hua; Wang, Fang; Bu, Peng-Bin; Lin, Sheng; Zhu, Cheng-Gen; Li, Yu-Huan; Jiang, Jian-Dong; Shi, Jian-Gong



Valproic acid inhibits invasiveness in bladder cancer but not in prostate cancer cells.  


Histone deacetylase inhibitors (HDACIs) represent a promising new class of antineoplastic agents that affect proliferation, differentiation, and apoptosis in both solid and hematologic malignancies. In addition, HDACIs can alter the expression of at least one cellular adhesion molecule, the coxsackie and adenovirus receptor, in bladder cancer. Because HDACIs can increase expression of a known cellular adhesion molecule, we hypothesized that migration and/or invasion may also be affected. We evaluated this hypothesis using valproic acid (VPA), a commonly prescribed anticonvulsant recently shown to have potent HDACI activity, in the bladder cancer cell lines T24 TCC-SUP, HT1376, and RT4. Analyses of cell migration and invasion were both qualitative (fluorescent microscopy) and quantitative (static and dynamic migration/invasion assays). Our results show that acute VPA treatment (72 h) causes a dose-dependent decrease in invasion for all bladder cancer cell lines, except RT4, a noninvasive papilloma. Migration, in contrast, was not affected by VPA treatment. The inhibitory effect of VPA may be cancer type-specific, because there was no difference in invasion between treated and untreated prostate cancer cell lines LNCaP, PC3, and DU145. Furthermore, when administered chronically (34 days), VPA significantly inhibits growth of T24t tumor xenografts. Our data suggest that VPA exerts some of its antineoplastic effects by inhibiting invasion as well as tumor growth, and thus it may represent a novel adjuvant strategy for patients at high risk of recurrence and/or progression of muscle invasive bladder cancer. PMID:16868035

Chen, Chien-Lun; Sung, Jennifer; Cohen, Michael; Chowdhury, Wasim H; Sachs, Markus D; Li, Ying; Lakshmanan, Yegappan; Yung, Benjamin Y M; Lupold, Shawn E; Rodriguez, Ronald



Diabetogenic effects of the most prevalent enteroviruses in Finnish sewage.  


Common enterovirus infections appear to initiate or facilitate the pathogenetic processes leading to type 1 diabetes (T1D) and also sometimes precipitate the clinical disease. We have recently demonstrated that (1) enterovirus-positive islet cells were seen on postmortem pancreatic specimens of several T1D patients but not in the corresponding samples of nondiabetic controls, and (2) several different enteroviruses can be associated with T1D. Enterovirus infections are transmitted from person to person by fecal-oral or respiratory routes, which means that infections usually start from the respiratory or gastrointestinal mucosa. Regardless of the clinical symptoms of the disease, viral replication continues in the submucosal lymphatic tissue for several weeks, up to a couple of months, and during that time the virus is excreted into the feces and translocated to the environment. Monitoring of sewage samples for enteroviruses can be used as a tool in epidemiologic studies of enterovirus. Finland has successfully used environmental control data in poliovirus surveillance for decades. About 24 samples have been collected annually from the Helsinki region, which covers about 20% of the population. In the present study, we have reanalyzed the sewage samples of the years 1993-2004 for nonpolio enteroviruses by inoculating them into five different continuous cell lines known to cover a wide range of serotypes. Isolated strains were identified by RT-PCR and VP1 sequencing. The most commonly detected serotypes were coxsackie B viruses (CBV1-5) and echoviruses (E6, 7, 11, 25, 30). Diabetogenic effects of the most prevalent enterovirus serotypes were studied in primary human beta cells. PMID:19120297

Klemola, Paivi; Kaijalainen, Svetlana; Ylipaasto, Petri; Roivainen, Merja



Effect of HLA genotype or CTLA-4 polymorphism on cytokine response in healthy children.  


Type 1 diabetes (T1D) is considered to be a T-cell-mediated autoimmune disease in which genetic predisposition is affected by HLA class II alleles and polymorphisms in cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) gene. We tested the hypothesis whether these T1D-related gene polymorphisms modulate cytokine response and thus contribute to the development of autoimmunity. The study includes 67 non-diabetic children, typed for HLA class II alleles and CTLA-4 polymorphisms (+49A/G, CT60A/G, CTBC217_1C/T). We measured cytokine secretion of peripheral blood mononuclear cells after stimulation with tetanus toxoid (TT), polio virus, coxsackie virus B4, pertussis toxin (PT) and phytohemagglutinin (PHA). We saw higher IL-13 response to TT in individuals with DR3-DQ2 haplotype (P = 0.002). HLA class II protective haplotype, DR2-DQ6, showed association with increased production of IFN-gamma (P < 0.001) and IL-2 (P = 0.005) in response to polio virus. In children with the autoimmunity-related homozygous genotypes CTLA-4 +49G/G, CT60G/G and CTBC217_1T/T, we found enhanced PT- and PHA-induced IFN-gamma production (P < 0.05). The cytokine responses to studied antigens were weakly modified by HLA class II risk haplotypes, and children with T1D-associated HLA risk haplotypes are not specifically inclined to develop an immune response in general. Higher IFN-gamma and IL-2 response to enterovirus in children with HLA class II protective haplotype DR2-DQ6 could be of importance in the protection from T1D-associated enterovirus infections. All autoimmunity related CTLA-4 polymorphisms were associated with enhanced IFN-gamma. This suggests impaired downregulation of cellular immunity by these CTLA-4 polymorphisms. PMID:18782261

Walldén, J; Ilonen, J; Roivainen, M; Ludvigsson, J; Vaarala, O



Intra-testicular injection of adenoviral constructs results in Sertoli cell-specific gene expression and disruption of the seminiferous epithelium.  


Spermatogenesis is a complex process that cannot be modelled in vitro. The somatic Sertoli cells (SCs) within the seminiferous tubules perform a key role in supporting maturation of germ cells (GCs). Progress has been made in determining what aspects of SC function are critical to maintenance of fertility by developing rodent models based on the Cre/LoxP system; however, this is time-consuming and is only applicable to mice. The aim of the present study was to establish methods for direct injection of adenoviral vectors containing shRNA constructs into the testis as a way of inducing target-selective knock-down in vivo. We describe here a series of experiments using adenovirus expressing a green fluorescent protein (GFP) transgene. Injection via the efferent ductules resulted in SC-specific expression of GFP; expression levels paralleled the amount of infective viral particles injected. At the highest doses of virus seminiferous tubule architecture were grossly disturbed and immune cell invasion noted. At lower concentrations, the expression of GFP was variable/negligible, the seminiferous tubule lumen was maintained but stage-dependent GC loss and development of numerous basal vacuoles was observed. These resembled intercellular dilations of SC junctional complexes previously described in rats and may be a consequence of disturbances in SC function due to interaction of the viral particles with the coxsackie/adenovirus receptor that is a component of the junctional complexes within the blood testis barrier. In conclusion, intra-testicular injection of adenoviral vectors disturbs SC function in vivo and future work will therefore focus on the use of lentiviral delivery systems. PMID:18955374

Hooley, R P; Paterson, M; Brown, P; Kerr, K; Saunders, P T K



Search for Coxsackievirus B3 RNA in idiopathic dilated cardiomyopathy using gene amplification by polymerase chain reaction.  


A polymerase chain reaction (PCR) amplification assay was developed to detect Coxsackievirus B3 ribonucleic acid (RNA) in blood and myocardial tissue of explanted hearts from 40 patients who underwent cardiac transplantation and in 1 normal heart. Twenty-one patients were affected by idiopathic dilated cardiomyopathy of different duration and 19 by coronary artery disease. Coxsackievirus B3 in vitro infected Vero cells and cells infected by related human enteroviruses (Coxsackievirus B2, B4, and poliovirus 1) were used as reaction controls. PCR was performed using 4 pairs of primers homologous to Coxsackie-virus B3 sequences. Three sets were located in regions of the genome conserved at nucleotide level between several enterovirus species (replicase gene, 5' noncoding region), while one was located in a Coxsackievirus B3-specific region (VP1 gene). Total RNA was prepared by acid guanidinium isothiocyanate extraction from tissue stored frozen at -80 degrees C. One microgram of total RNA was retrotranscribed with either antisense primer or with random hexanucleotide primers and then subjected to 40 cycles of amplification. PCR products were separated by electrophoresis on a 10% polyacrylamide gel, electrotransferred to a nylon membrane and then hybridized to oligonucleotide probes specific for the coxsackievirus B3 genome radiolabeled with radioactive isotope of phosphorous. All pairs of primers yielded specific amplification products when tested on Coxsackievirus B3-infected Vero cells, with a sensitivity of 1 infected cell out of 10(5) to 10(6) cells starting from 1 microgram total RNA. Primer sets for regions of Coxsackievirus B3 genome highly conserved between related enteroviral species gave positive amplification also when challenged with RNA from cells infected by Coxsackievirus B2, B4 and poliovirus 1. PMID:1311139

Grasso, M; Arbustini, E; Silini, E; Diegoli, M; Percivalle, E; Ratti, G; Bramerio, M; Gavazzi, A; Vigano, M; Milanesi, G



Adenovirus type-35 vectors block human CD4+ T-cell activation via CD46 ligation.  


Recombinant adenoviruses (rAds) based on types 5 (rAd5) and 35 (rAd35) have emerged as important vaccine delivery vectors in clinical testing for a variety of pathogens. A major difference between these vectors is their binding to cellular receptors used for infection. Whereas rAd5 binds coxsackie-adenovirus receptor (CAR), rAd35 binds the complement regulatory protein CD46. Although rAd35 infected and phenotypically matured human blood dendritic cells (DCs) more efficiently than rAd5, we show here that rAd35 markedly suppressed DC-induced activation of naive CD4(+) T cells. rAd35 specifically blocked both DCs and anti-CD3/CD28 mAb-induced naive T-cell proliferation and IL-2 production. This effect was also observed in CD4(+) memory T cells but to a lesser extent. The suppression occurred by rAd35 binding to CD46 on T cells and was independent of infection. CD46 engagement with mAb mimicked the effects of rAd35 and also led to deficient NF-?B nuclear translocation. In contrast, rAd5 and rAd35 vectors with ablated CD46 binding did not inhibit T-cell activation. Our findings provide insights into the basic biology of adenoviruses and indicate that CD46 binding may have an impact on the generation of primary CD4(+) T-cell responses by Ad35. PMID:21502499

Adams, William C; Gujer, Cornelia; McInerney, Gerald; Gall, Jason G D; Petrovas, Constantinos; Karlsson Hedestam, Gunilla B; Koup, Richard A; Loré, Karin



Adenovirus type-35 vectors block human CD4+ T-cell activation via CD46 ligation  

PubMed Central

Recombinant adenoviruses (rAds) based on types 5 (rAd5) and 35 (rAd35) have emerged as important vaccine delivery vectors in clinical testing for a variety of pathogens. A major difference between these vectors is their binding to cellular receptors used for infection. Whereas rAd5 binds coxsackie-adenovirus receptor (CAR), rAd35 binds the complement regulatory protein CD46. Although rAd35 infected and phenotypically matured human blood dendritic cells (DCs) more efficiently than rAd5, we show here that rAd35 markedly suppressed DC-induced activation of naive CD4+ T cells. rAd35 specifically blocked both DCs and anti-CD3/CD28 mAb-induced naive T-cell proliferation and IL-2 production. This effect was also observed in CD4+ memory T cells but to a lesser extent. The suppression occurred by rAd35 binding to CD46 on T cells and was independent of infection. CD46 engagement with mAb mimicked the effects of rAd35 and also led to deficient NF-?B nuclear translocation. In contrast, rAd5 and rAd35 vectors with ablated CD46 binding did not inhibit T-cell activation. Our findings provide insights into the basic biology of adenoviruses and indicate that CD46 binding may have an impact on the generation of primary CD4+ T-cell responses by Ad35.

Adams, William C.; Gujer, Cornelia; McInerney, Gerald; Gall, Jason G. D.; Petrovas, Constantinos; Hedestam, Gunilla B. Karlsson; Koup, Richard A.; Lore, Karin



Animal models used to examine the role of the environment in the development of autoimmune disease: findings from an NIEHS Expert Panel Workshop.  


Autoimmunity is thought to result from a combination of genetics, environmental triggers, and stochastic events. Environmental factors, such as chemicals, drugs or infectious agents, have been implicated in the expression of autoimmune disease, yet human studies are extremely limited in their ability to test isolated exposures to demonstrate causation or to assess pathogenic mechanisms. In this review we examine the research literature on the ability of chemical, physical and biological agents to induce and/or exacerbate autoimmunity in a variety of animal models. There is no single animal model capable of mimicking the features of human autoimmune disease, particularly as related to environmental exposures. An objective, therefore, was to assess the types of information that can be gleaned from the use of animal models, and how well that information can be used to translate back to human health. Our review notes the importance of genetic background to the types and severity of the autoimmune response following exposure to environmental factors, and emphasizes literature where animal model studies have led to increased confidence about environmental factors that affect expression of autoimmunity. A high level of confidence was reached if there were multiple studies from different laboratories confirming the same findings. Examples include mercury, pristane, and infection with Streptococcus or Coxsackie B virus. A second level of consensus identified those exposures likely to influence autoimmunity but requiring further confirmation. To fit into this category, there needed to be significant supporting data, perhaps by multiple studies from a single laboratory, or repetition of some but not all findings in multiple laboratories. Examples include silica, gold, TCE, TCDD, UV radiation, and Theiler's murine encephalomyelitis virus. With the caveat that researchers must keep in mind the limitations and appropriate applications of the various approaches, animal models are shown to be extremely valuable tools for studying the induction or exacerbation of autoimmunity by environmental conditions and exposures. PMID:22748431

Germolec, Dori; Kono, Dwight H; Pfau, Jean C; Pollard, K Michael



[Relation between microbiological monitoring and the risk of hospital infection in obstetrical departments].  


1. The increased microbiological control of the total work process at maternity wards enables the timely notice of the contamination location. 2. A direct result of such control is an improvement of hygienic-technological procedures, disinfection and sterilisation at maternity wards. 3. A favourable epidemiological situation was registered during a two-year period following the epidemic (1986-1987) caused by the Coxsackie B virus with respect to a two-year investigation period preceding the epidemic (1984-1985). 4. During the two-year investigation period preceding the epimic 1,942 microbiological controls were performed at the Ginealogical-Obstetrics Clinic in Sarajevo. During the same period following the epidemic 3.321 microbiological controls were performed which is an increase of 1.7 times. 5. Increased number of controls influenced the improvement of the hygienic regime, and the basic parameters were: decreased prevalence of dangerous bacteria species, increased number of sterile swabs and a decreased number of pathogenous isolates. 6. The evaluation of hygiene and epidemiological risk of the medical equipment based on isolated pathogenous carriers prior and after the epidemic shows a significant difference (P less than 0.01). 7. There is a significant statistical difference (P less than 0.01) concerning the hygienic conditions of working clothes with respect to isolated pathogenous carriers before and after the epidemic. 8. Increased microbiological supervision resulted in the elimination of risks associated with working and floor surfaces since findings during the 1986-1987. period in comparison with the 1984-1985. period show significant statistical results with respect to pathogenous isolates (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2626070

Basi?, F; Simi?, S; Puvaci?, Z; Simi?, E; Milisi?, J



The combination of an oxygen-dependent degradation domain-regulated adenovirus expressing the chemokine RANTES/CCL5 and NK-92 cells exerts enhanced antitumor activity in hepatocellular carcinoma  

PubMed Central

Oncolytic adenoviruses are modified based on adenovirus serotype 5 (Ad5), which belongs to subgroup C and depends on Coxsackie-adenovirus receptor (CAR) to recognize target cells. However, expression of CAR is generally low or lost in certain tumors including hepatocellular carcinoma (HCC). By contrast, CD46 is highly expressed in various types of malignant tumor cells. Therefore, we constructed an adenovirus vector expressing the human RANTES/CCL5 gene regulated by oxygen-dependent degradation domain (ODD) and analyzed its antitumor effects in vitro and in vivo. The human RANTES/CCL5 gene was fused with ODD by PCR and the recombinant oncolytic adenovirus containing RANTES-ODD, SG511-CCL5-ODD, was constructed by the Gateway system, which infected cells by binding CD46. Viral replication experiments were performed to evaluate the selective replication ability of SG511-CCL5-ODD. RANTES expression was determined by ELISA. The chemotactic test was used to analyze the ability of the expressed RANTES to recruit NK92 cells. The antitumor effects of SG511-CCL5-ODD were examined in HCC xenografts in nude mice. A chimeric oncolytic adenovirus, SG511-CCL5-ODD, was constructed successfully. Cells infected with the recombinant virus were able to express RANTES selectively in different environments controlled by ODD and the expressed RANTES was able to recruit NK92 cells by its chemotactic effect in vitro and improve the anticancer immune response in HCC xenografts in nude mice. The chimeric adenovirus SG511-CCL5-ODD highly expressed the RANTES-ODD fusion gene in the hypoxia of HCC under the control of the ODD and effectively attracted NK92 cells and a high number of immunocytes. These factors had complementary advantages and, in combination, exerted enhanced antitumor efficacy.




The pseudopeptide HB-19 binds to cell surface nucleolin and inhibits angiogenesis  

PubMed Central

Background Nucleolin is a protein over-expressed on the surface of tumor and endothelial cells. Recent studies have underlined the involvement of cell surface nucleolin in tumor growth and angiogenesis. This cell surface molecule serves as a receptor for various ligands implicated in pathophysiological processes such as growth factors, cell adhesion molecules like integrins, selectins or laminin-1, lipoproteins and viruses (HIV and coxsackie B). HB-19 is a synthetic multimeric pseudopeptide that binds cell surface expressed nucleolin and inhibits both tumor growth and angiogenesis. Methodology/principal findings In the present work, we further investigated the biological actions of pseudopeptide HB-19 on HUVECs. In a previous work, we have shown that HB-19 inhibits the in vivo angiogenesis on the chicken embryo CAM assay. We now provide evidence that HB-19 inhibits the in vitro adhesion, migration and proliferation of HUVECs without inducing their apoptosis. The above biological actions seem to be regulated by SRC, ERK1/2, AKT and FAK kinases as we found that HB-19 inhibits their activation in HUVECs. Matrix metalloproteinases (MMPs) play crucial roles in tumor growth and angiogenesis, so we investigated the effect of HB-19 on the expression of MMP-2 and we found that HB-19 downregulates MMP-2 in HUVECs. Finally, down regulation of nucleolin using siRNA confirmed the implication of nucleolin in the biological actions of these peptides. Conclusions/significance Taken together, these results indicate that HB-19 could constitute an interesting tool for tumor therapy strategy, targeting cell surface nucleolin.



Distribution of enteroviruses in hospitalized children with hand, foot and mouth disease and relationship between pathogens and nervous system complications  

PubMed Central

Background To explore the relationship between enteroviruses and hospitalized children with hand, foot and mouth disease (HFMD) complicated with nervous system disease. 234 hospitalized HFMD patients treated in Shengjing Hospital, Liaoning Province were analyzed retrospectively. Based on the presence and severity of nervous system disease, the patients were grouped as follows: general patients, severely ill patients, critically ill patients and fatal patients. Based on the detected pathogen, the patients were grouped as follows: Enterovirus 71 (EV71) infection, coxsackie A16 (CA16) infection and other enterovirus (OE) infection. Results Of the 423 hospitalized patients, most were admitted in July 2010(129/423, 30.5%). Enteroviruses were detected in 177(41.8%). 272/423 patients were male (64.3%), and fatal patients had the greatest proportion of male patients (p < 0.05). EV71 infection was found in 89/423 patients (21%). CA16 infection was detected in 8/423 patients (16.1%). Compared to group CA16, patients in group EV71 were hospitalized earlier, and the duration of hospitalization was longer (p < 0.05). Of the 92 patients with nervous system damage, 65 were infected with EV71 and 19 were infected with CA16. Among these CA16 infected patients, 2 had brainstem encephalitis and 1 had AFP. There were more patients with nervous system dysfunction in group EV71 than in groups CA16 or OE (p < 0.05). The 5 fatalities all occurred in group EV71 patients (p < 0.05). Infection with EV71 was most likely to cause neurogenic pulmonary edema (p < 0.05). Patients in group EV71 had a higher rate of suffering from coma and limb movement disorder than patients in groups CA16 or OE (p < 0.05). Conclusion The disease progresses faster in EV71-infected HFMD patients. These patients are more likely to suffer nervous system damage, neurogenic pulmonary edema, severe sequelae or death. CA16 and other enteroviruses can also cause HFMD with severe nervous system complications.



Influence of Fiber Detargeting on Adenovirus-Mediated Innate and Adaptive Immune Activation  

PubMed Central

The major adenovirus (Ad) capsid proteins hexon, penton, and fiber influence the efficiency and tropism of gene transduction by Ad vectors. Fiber is the high-affinity receptor binding protein that serves to mediate cell attachment in vitro when using coxsackie-adenovirus receptor (CAR)-containing cell lines. This contrasts with transduction efficiency in macrophages or dendritic cells that lack high concentrations of CAR. To determine how fiber influences gene transduction and immune activation in a murine model, we have characterized Ad type 5 (Ad5) vectors with two classes of chimeric fiber, CAR binding and non-CAR binding. In a systemic infection, Ad5 fiber contributes to DNA localization and vector transduction in hepatic tissue. However, the majority of vector localization is due to Ad5 fiber-specific functions distinct from CAR binding. CAR-directed transduction occurs but at a modest level. In contrast to CAR binding vectors, the F7 and F7F41S non-CAR-binding vectors demonstrate a 2-log decrease in hepatic transduction, with a 10-fold decrease in the amount of vector DNA localizing to the hepatic tissue. To characterize the innate response to early infection using fiber chimeric vectors, intrahepatic cytokine and chemokine mRNAs were quantified 5 hours postinfection. Tumor necrosis factor alpha mRNA levels resulting from Ad5 fiber infections were elevated compared to viruses expressing serotype 7 or 41 fiber. Levels of chemokine mRNA (gamma interferon-inducible protein 10, T-cell activation gene 3, and macrophage inflammatory protein 1?) were 10- to 20-fold higher with CAR binding vectors (Ad5 and F41T) than with non-CAR-binding vectors (F7 and F7F41S). In spite of quantitative differences in vector localization and innate activation, fiber pseudotyping did not significantly change the outcome of anti-Ad adaptive immunity. All vectors were cleared with the same kinetics as wild-type Ad5 vectors, and each induced neutralizing antibody. Although non-CAR-binding vectors were impaired in transduction by nearly 2 orders of magnitude, the level of antitransgene immunity was the same for each of the vectors. Using primary bone marrow-derived macrophages and dendritic cells, we demonstrate that transduction, induction of cytokine/chemokine, and phenotypic maturation of these antigen-presenting cells are independent of fiber content. Our data support a model where fiber-mediated hepatic localization enhances innate responses to virus infection but minimally impacts on adaptive immunity.

Schoggins, John W.; Nociari, Marcelo; Philpott, Nicola; Falck-Pedersen, Erik



Comparative proteomics of the Mycobacterium leprae binding protein myelin P0: its implication in leprosy and other neurodegenerative diseases.  


Mycobacterium leprae, the causative agent of leprosy invades Schwann cells of the peripheral nerves leading to nerve damage and disfigurement, which is the hallmark of the disease. Wet experiments have shown that M. leprae binds to a major peripheral nerve protein, the myelin P zero (P0). This protein is specific to peripheral nerve and may be important in the initial step of M. leprae binding and invasion of Schwann cells which is the feature of leprosy. Though the receptors on Schawann cells, cytokines, chemokines and antibodies to M. leprae have been identified the molecular mechanism of nerve damage and neurodegeneration is not clearly defined. Recently pathogen and host protein/nucleotide sequence similarities (molecular mimicry) have been implicated in neurodegenerative diseases. The approach of the present study is to utilise bioinformatic tools to understand leprosy nerve damage by carrying out sequence and structural similarity searches of myelin P0 with leproma and other genomic database. Since myelin P0 is unique to peripheral nerve, its sequence and structural similarities in other neuropathogens have also been noted. Comparison of myelin P0 with the M. leprae proteins revealed two characterised proteins, Ferrodoxin NADP reductase and a conserved membrane protein, which showed similarity to the query sequence. Comparison with the entire genomic database ( by basic local alignment search tool for proteins (BLASTP) and fold classification of structure-structure alignment of proteins (FSSP) searches revealed that myelin P0 had sequence/structural similarities to the poliovirus receptor, coxsackie-adenovirus receptor, anthrax protective antigen, diphtheria toxin, herpes simplex virus, HIV gag-1 peptide, and gp120 among others. These proteins are known to be associated directly or indirectly with neruodegeneration. Sequence and structural similarities to the immunoglobin regions of myelin P0 could have implications in host-pathogen interactions, as it has homophilic adhesive properties. Although these observed similarities are not highly significant in their percentage identity, they could be functionally important in molecular mimicry, receptor binding and cell signaling events involved in neurodegeneration. PMID:15019586

Vardhini, Deena; Suneetha, Sujai; Ahmed, Niyaz; Joshi, D S M; Karuna, S; Magee, X; Vijayalakshmi, D S R; Sridhar, V; Karunakar, K V; Archelos, Juan J; Suneetha, Lavanya M