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作 者:倪兵[1] 王希良[2] 王莉[1] 赵光宇[2] 石辛甫[2] 张松乐[2] 张良艳[2] 罗德炎[2] 黎万玲[1] 姜曼[1] 毛丽伟[1] 何仰东[1] 肖宇[1] 高闻达 吴玉章[1]
机构地区:[1]第三军医大学全军免疫学研究所 [2]军事医学科学院微生物流行病学研究所免疫室 [3]BethIsrael Deaconess MedicalCenter,Harvard MedicalSchool,99 Brookline Ave.,Boston,MA02215
出 处:《免疫学杂志》2005年第6期433-437,共5页Immunological Journal
基 金:国家重大基础研究计划("973"项目);SARS预防基础研究基金(2003CB514108);自然科学基金重大研究项目(30490240);杰出青年基金(30325020);"863"计划(2003AA208224;2003AA208212;2003AA208218)资助
摘 要:目的探讨马抗SARSCoV中和性抗体是否对SARSCoV感染有防治作用。方法用SARSCoV(BJ01株)免疫马匹,从免疫马血清中制备IgGs及其F(ab’)2片段。通过细胞病变法(CPE)、MTT法和空斑形成实验(PFU)等方法,在体外培养的VeroE6细胞中检测F(ab’)2片段对SARSCoV感染的预防性和治疗性作用。再在Balb/c小鼠体内模型中,用CPE法和定量RTPCR方法检测该抗体的保护性效应。结果CPE、MTT和PFU三种方法证实来自三匹马的血清F(ab’)2的中和滴度均达到1∶1600以上。体内实验证实,50μg的F(ab’)2片段可以完全中和1×104TCID50SARSCoV。结论马抗SARSCoV抗体在体内外均能有效地中和SARSCoV和预防其感染宿主细胞,为马抗体将来在大的灵长类动物或人体内进行实验提供实验依据。Objective To investigate the effects of equine neutralizing antibody on SARS-CoV infection in primate cells. Methods IgGs and their pepsin-dlgested F(ab')2 fragments were prepared from the sera of three horses inoculated with purified SARS-CoV (BJ-01 strain). The protective effects of the F(ab')2 fragments against SARS-CoV infection was determined in cultured Vere E6 cells by cytopathic effect (CPE), MTT, and plaque-forming assays, as wellas in a Balb/c mouse model by CPE and quantitative RT-PCR assays. Results The neutra-lization titres of F(ab')2 from three horses all reached at least 1:1600 and 50/tg of the F(ab')2 fragments could completely neutralize 1×10^4 TCID50 SARS-CoV in vivo. We also observed that F(ab')2 against BJ-01 strain could protect cells from infection of the variant GZ-01 strain in vitro and in vivo as well. Conclusion The antibody against SARS-CoV raised in horses could neutralize SARS-CoV and prevent its infection in host efficaciously. Our work provided experimental support for testing the protective equine immunoglobulin in future large primate or human trial.
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