关键词:肝炎病毒组,丙型;基因免疫
【摘 要】 目的 研究丙型肝炎病毒(HCV)核心(C)基因免疫在诱生特异性细胞免疫应答中的作用。方法 将包含HCVC基因片段的重组真核表达质粒pcDNA HCV C,在证实其可以在小鼠骨髓瘤细胞SP2/0(H-2d )中表达之后,注射BALB/c小鼠股四头肌。ELISA法检测血清中抗体水平;3 H-TdR掺入法测定免疫小鼠脾细胞特异性增殖能力;51 Cr释放法检测免疫小鼠细胞毒性T淋巴细胞(CTLs)体外杀伤功能。结果 8只免疫小鼠血清中均出现了HCVC抗体,免疫组小鼠脾细胞增殖能力明显高于对照组(P<0.001),增殖指数为6.10,CTLs体外特异性杀伤率为63.13%,也高于对照组(P<0.01)。结论 HCVC基因免疫可能是防治HCV感染的有效方法。
Immune responses torecombinant expression plasmid encoding
hepatitis C virus core antigen
FENG Zhihua, ZHOU Yongxing, JIA Zhansheng, et al.
Department of Infectious Diseases, Tangdu Hospital, The Fourth Military MedicalUniversity, Xi′an 710038
【Abstract 】 Objective Tostudy the feasibility of DNA immunization with recombinant expression plasmid containinghepatitis C virus (HCV) core gene against HCV infection. Methods Therecombinant plasmid containing HCV C gene was constructed and expressed transiently byusing lipofectamine in the mouse SP2/0 cells. After appraisal and purification, theseplasmid and control DNA were directly injected intramuscularly into BALB/c mice, HCV Cspecific antibody in the sera was checked with ELISA. Proliferative responses of spleencells from experimental and control mice were determined by 3 H thymidineincorporation and cytotoxicity T lymphocyte (CTL) activity was measured by standard 4-h 51 Crrelease assays. Results All immunized mice developed anti-HCV coreantibodies and this antibody was dose and immune number dependent. Lymphoproliferativeresponses for HCV core antigen were higher in immunized mice than in control mice(P<0.05), stimulation indices (SI) were 2.42~6.10. CTLs might induce approximately63.13% lysis of pcDNA HCV C transfected SP2/0 target cells. Conclusion Itis suggested that HCV C gene immunization is a promising method of vaccination against HCVinfection.
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