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Establishment of animal model for potency evaluationof inactivated sars virus experimental vaccine

Journal of Microbiology and Immunology 2005年第1期3卷 53-60页

作者： GUANMUDONG QIAN YANKONG WENXUELIU LIHINGYANG JUNZHIWANG YONGXINYU YAOLONGSHU ZHENGWANG WEIDONGYIN QINGYUZHU HAIFAZHENG NationalInstitutefortheControlofPharmaceuticalandBiologicalProducts BeijingP.R.China BeijingJindikeBio-TechnologyInstitute BeijingP.R.China SinovacBiotechCo.Ltd.39ShangdiXiRoad HaidianDistrictBeijingP.R.China MicrobiologyandEpidemicInstituteofAcademyofMilitaryMedicalScience BeijingP.R.China HuatesenGeneCo.Ltd BeijingP.R.China

The purpose of this study was to test the effectiveness of source virus strain for the manufacture of the inactivated sars virus vaccine, and establish an experimental method and preliminary standard for potency evaluation. Mice were divided into groups for being immunized with corresponding serially diluted experimental sars virus inactivated vaccine. And the rabbits were immunized with undiluted vaccine. Challenge assay was conducted with a heterologous sars virus. And the neutralization antibody was determined with plaque reduction neutralization test (PRNT), to which the neutralization antibody in the convalescent serum of sars patients was compared. The experimental vaccine viral strains were proved to be suitable for manufacturing the vaccine. Mice immunized by vaccines of serial dilutions were able to elicit neutralizing antibody. The antibody titer from mice immunized with the undiluted vaccine could reach up to 1∶495.2, while those of rabbits immunized with the undiluted vaccine could reach a GMT of 55.0-79.9. The capability of the antibody to neutralize the virus from Guangdong is more efficient than that from Beijing. The GMT of neutralizing antibody in sars convalescents living in south and north China ranged from 50.12 to 54.95, and the titers of convalescents from north China were higher than those from south China. Mice and rabbits used as the model for evaluation of potency are of sensitivity, and the test is of reproducibility. The candidate challenge viral strains showed a relatively consistent effect on evaluating antibodies produced by various batches and different vaccine-source strains, hence they can be used to evaluate potency of the vaccine. The method for testing the vaccine potency and the evaluation standard was established preliminarily.

关键词： sars virus Inactivated vaccine Neutralizing antibody Potency assay Plaque reduction neutralization test(PRNT)

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Humoral immune responses in rabbits induced by an experimental inactivated severe acute respiratory syndrome coronavirus vaccine prepared from F69 strain

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Chinese Medical Journal 2004年第11期117卷 1625-1629页

作者：张传海郭中敏郑焕英陆家海王一飞鄢心革赵勇杜雄威张欣方苓凌文华戚树源余新炳钟南山 South China Institute of Botany Graduate School of Chinese Academy of Sciences Guangzhou 510650 China Department of Medical Statistics and Epidemiology School of Public Health Sun Yat-sen University Guangzhou 510080 China Center for Disease Control and Prevention of Guangdong Province Guangzhou 510300 China Biomedicine Research and Development Center Jinan University Guangzhou 510632 China Department of Medical Statistics and Epidemiology School of Public Health Sun Yat-sen University Guangzhou 510080 China Guangzhou Institute of Respiratory Diseases Guangzhou Medical College Guangzhou 510120 China

BACKGROUND: The etiologic agent of severe acute respiratory syndrome (sars) has been confirmed to be a novel coronavirus (CoV), namely sars-CoV. Developing safe and effective sars-CoV vaccines is essential for us to prevent the possible reemergence of its epidemic. Previous experiences indicate that inactivated vaccine is conventional and more hopeful to be successfully developed. Immunogenicity evaluation of an experimental inactivated sars-CoV vaccine in rabbits was conducted and reported in this paper. METHODS: The large-scale cultured sars-CoV F69 strain was inactivated with 0.4% formaldehyde and purified, then used as the immunogen combined with Freund's adjuvant. Eight adult New Zealand rabbits were immunized four times with this experimental inactivated vaccine. Twelve sets of rabbit serum were sampled from the third day to the seventy-fourth day after the first vaccination. The titers of specific anti-sars-CoV IgG antibody were determined by indirect enzyme-linked immunosorbent assay, and the neutralizing antibody titers were detected with micro-cytopathic effect neutralization test. RESULTS: Rapid and potent humoral immune responses were induced by the inactivated sars-CoV vaccine in all the eight test rabbits. Titers of both specific IgG antibody and neutralizing antibody peaked at about six weeks after first vaccination, with the maximum value of 1:81 920 and 1:20 480, respectively. After that, serum antibody levels remained at a plateau or had a slight decrease, though two boosters were given in the succedent 4 to 5 weeks. Cross neutralization response existed between sars-CoV F69 strain and Z2-Y3 strain. CONCLUSIONS: The inactivated sars-CoV vaccine made from F69 strain owns strong immunogenicity, and the cross neutralization response between the two different sars-CoV strains gives a hint of the similar neutralizing epitopes, which provide stable bases for the development of inactivated sars-CoV vaccines.

关键词： Animals Antibodies, Viral Immunoglobulin G Neutralization Tests Rabbits Research Support, Non-U.S. Gov't sars virus Vaccines, Inactivated Viral Vaccines

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Inhibiting severe acute respiratory syndrome-associated coronavirus by small interfering RNA

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Chinese Medical Journal 2003年第8期116卷 1262-1264页

作者：张仁礼郭中敏陆家海孟锦绣周灿权詹希美黄冰余新炳黄民潘兴华凌文华陈系古万卓越郑焕英鄢心革王一飞冉延超刘新健马俊鑫王承宇张必良 Sun Yat-Sen University Center for Disease Control and Prevention of Guangdong Province Jinan University Maxim Biotech Inc. The Quartermaster University of PLA Medical School of University of Massachusetts

OBJECTIVE: To evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (sars)-associated coronavirus replication, and to lay bases for the future clinical application of siRNA for the treatment of viral infectious diseases. METHODS: Vero-E6 cells was transfected with siRNA before sars virus infection, and the effectiveness of siRNA interference was evaluated by observing the cytopathic effect (CPE) on Vero-E6 cells. RESULTS: Five pairs of siRNA showed ability to reduce CPE dose dependently, and two of them had the best effect. CONCLUSION: siRNA may be effective in inhibiting sars-associated coronavirus replication.

关键词： Animals Cercopithecus aethiops RNA, Small Interfering sars virus Transfection Vero Cells virus Replication

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