Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum
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Keywords:
Infectivity
Antibody titer
Coxsackievirus
Cytopathic effect
Objective To observe the protective effect of momordicin on Coxsackievirus B3 infected rat cardiocyte and its mechanism. Methods Rat cardiocyte were prime cultured in vitro,then infected with Coxsackievirus B3,and the viral myocarditis model was duplicated,the protective effect of momordicin was analyzed by measuring median effective dose(ED50),percents of beat cells(PBC),cytopathic effect(CPE),myocardiocyte related enzymes and tissue culture infective dose(TCID50). Results A distinct protective effect of momordicin on CVB3 infected myocardiocyte was found(ED50=0.045 mg/mL).At the concentration of 80,40,20,10 μg/mL,distinct effects of enhancing PBC,reduction of CPE(cytopathic effect) and replication of CVB3,and at the concentration of 80,40 μg/mL,LDH and AST were reduced remarkably. Conclusion Momordicin has a strong protective effect on CVB3 infected cardiocyte.
Coxsackievirus
Cytopathic effect
ED50
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Bablanian, Rostom (The Rockefeller University, New York, N.Y.), Hans J. Eggers, and Igor Tamm . Inhibition of enterovirus cytopathic effects by 2-(α-hydroxybenzyl)-benzimidazole. J. Bacteriol. 91: 1289–1294. 1966.—2-(α-Hydroxybenzyl)-benzimidazole (HBB), a specific inhibitor of enterovirus multiplication, markedly delayed the development of cytopathological changes induced by echovirus 12 or coxsackievirus B4 in monkey kidney cells, but did not prevent the ultimate degeneration of infected cells, even though virus multiplication was inhibited. The study of the development of viral cytopathic effects was facilitated by the use of antiviral immune serum, which restricted the infection to those cells which became infected by the inoculated virus and thereby established single-cycle conditions. With echovirus 12 and coxsackievirus B4 not all cells could be infected initially, even when cultures were inoculated at input multiplicities in excess of 100 plaque-forming units per cell.
Echovirus
Coxsackievirus
Cytopathic effect
Benzimidazole
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Hantaan virus
Marburg virus
Medical microbiology
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Objective To investigate the role of cardiotrophin-1 in cardiomyocyte infected by coxsackievirus,and then to explore whether the specific role works via STAT3 pathway.Methods Cardiomyocytes extracted from SD rat were infected by coxsackievirus B3 to establish cell-based viral myocarditis model.Culturing for 48 h,the cardiomyocytes were divided into 5 groups: Con,Virus,CT-1,AG490 and AG490+CT-1.After processing for 30 min,the level of phosphorylated STAT3 was detected by Western blot.CVB3-mediated myocytopathic effects were observed after co-culturing for further 12,24,36 h respectively,myocardial cell lesions were examined with LDH assay.Results After CVB3 infection,the beating rate decreased gradually and then stopped.Meanwhile the level of LDH increased significantly in Virus and AG490 groups compared with the Con group(P0.05).In Virus and AG490 groups,the level of LDH increased gradually compared with former time points(P0.05).In CT-1 group the phosphorylation of STAT3 was prominent compared with Con group(P0.05).The degree of cytopathic effect in CT-1 group decreased significantly compared with virus group.Consistently,the release of LDH in CT-1 group was sharply less than virus group(P0.05).The phosphorylation of STAT3 in AG490+CT-1 group decreased significantly,the cytopathic effect was visible,the level of LDH increased significantly compared with Con and CT-1 group(P0.05).There was no difference in comparison with Virus group(P0.05).Conclusion CT-1 can protect cardiac cells from injury caused by CVB3,and the protective effect can be blocked by AG490,an inhibitor of STAT3 phosphorylation,which conclude that the protective role of CT-1 is essentially mediated by STAT3 activation during the infection of CVB3.
Coxsackievirus
Cytopathic effect
Viral Myocarditis
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Hybridoma cell lines that secrete monoclonal antibodies which react with HeLa cell surface antigens were produced. The monoclonal antibodies prevented cytopathic effects caused by coxsackievirus B1 and significantly reduced the amounts of coxsackieviruses B1, B5, and B6 that absorb to HeLa cells. These antibodies did not protect the cells from poliovirus cytopathic effects, and they had no effect on the attachment of other picornaviruses to HeLa cells.
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HeLa
Cytopathic effect
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( S,S )-1,2-bis(5-methoxy-2-benzimidazolyl)-1,2-ethanediol showed antiviral activity in monolayer tissue culture systems against 55 strains of rhinovirus, three types of poliovirus, and strains of type A and B coxsackieviruses. Neither the compound nor any of the analogues tested showed virucidal activity. Its antiviral activity was not associated with interference with viral attachment to or penetration into the cell. At a concentration of 0.1 mg/ml, this group of compounds was generally nontoxic to WI-38, primary bovine kidney, and African green monkey kidney cells and had antiviral activity with 100% inhibition of virus-induced cytopathic effects (CPE). At antiviral levels, these compounds prevented CPE of up to 10 6 median tissue culture infective dose units of virus and completely inhibited formation of new infective virions. The compounds showed antiviral activity both prophylactically and therapeutically against rhinoviruses. Infected cultures could be cleared of CPE up to 90 hr after infection.
Rhinovirus
Cytopathic effect
Coxsackievirus
Clearance
Echovirus
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Infectivity
Vero cell
Virus quantification
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Objective:The compounded Qiangqi extract was used to investigate its antiviral activity in Vero cells as a specific inhibitor of Coxsackievirus B 3 (CVB 3) replication, and its antiviral mechanism was exploredMethods:Vero cells infected by CVB 3 were cultured with different concentration of the compounded Qiangqi extract for 48 hours, and its inhibitory effect on CVB 3 replication was evaluated with cell survival rates by MTT assay and 50% tissue culture infection dose (TCID 50 ) of culture supernatantResults: The compounded Qiangqi extract could significantly inhibit cytopathic effect (CPE) of CVB 3-infected Vero cells, increase survival rates of CVB 3infected cells in dose dependent manner and decrease viral titres of culture supernatantConclusion:It may be the antiviral mechanism of the compounded Qiangqi extract that it may kill viruses directly and inhibit the replication of CVB 3 in cells, without preventing from viral adsorption and penetration
Vero cell
Coxsackievirus
Cytopathic effect
MTT assay
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Coxsackievirus
Cytopathic effect
Hydrochloride
Group A
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A micro-focus reduction neutralization test (mFRNT) was evaluated as an alternative test to the ordinary plaque reduction neutralization test (PRNT) for the determination of dengue virus and Japanese encephalitis virus neutralizing antibody responses in persons receiving dengue vaccine. The 2 tests were similar in terms of titres and ability to detect seroconversion. Although the neutralizing antibody titres obtained by mFRNT were slightly lower than those given by PRNT, the differences were less than two-fold, indicating that mFRNT was reliable. Reproducibility of mFRNT was confirmed by 10 replicate tests using the same control serum. Therefore, mFRNT may be useful in large-scale investigations of neutralizing antibody levels, for example, in young children forming part of an immunization programme; it can be performed quickly and is economical, requiring only a small volume of sera.
Dengue vaccine
Seroconversion
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