C3d-P28对基因免疫诱导的HBV特异性细胞免疫应答的调节作用  

P28 derived from complement C3d regulates specific cellular immunoresponse against HBV induced by gene immunization

作　　者：王立新[1] 关庆东[1] 徐薇[1] 王缨[1] 熊思东[1] 

机构地区：[1]复旦大学上海医学院免疫学系上海基因免疫与疫苗研究中心

出　　处：《中国免疫学杂志》2004年第7期443-447,共5页Chinese Journal of Immunology

基　　金：国家杰出青年科学基金研究计划 (3 992 5 0 3 1);国家重点基础研究发展计划 (2 0 0 1CB5 10 0 0 6)资助项目

摘　　要：目的 :观察补体C3d P2 8对基因免疫诱导的HBV特异性细胞免疫应答的调节作用 ,为增强基因免疫效果寻求新方法。方法 :将质粒pVAON33 S2 S质粒 (仅含HBV preS2 S编码基因 )和pVAON33 S2 S P2 8 4质粒 (含HBV preS2 S和 4拷贝C3d P2 8的编码基因 )以肌肉注射法对BALB C小鼠实施基因免疫 (10 0 μg 10 0 μl 只 ) ,并以空载质粒为对照。定期采集免疫小鼠血清、ELISA法检测其中特异性抗HBs IgG及其亚型的水平 ;免疫小鼠脾细胞经特异性抗原 (HBsAg)刺激后 ,采用3H TdR掺入法、半定量RT PCR法、同位素释放法分别检测其特异性淋巴细胞增殖活性、IL 4和IFN γ基因表达的水平、CTL杀伤活性。结果 :pVAON33 S2 S P2 8 4质粒基因免疫诱导的特异性淋巴细胞增殖活性、抗HBs IgG水平以及特异性CTL杀伤活性均明显高于pVAON33 S2 S质粒 ;C3d P2 8未改变基因免疫诱导的抗HBs IgG各亚型水平的格局 ,同时增强IgG1、IgG2a、IgG2b的水平 ;pVAON33 S2 S P2 8 4质粒诱导的IL 4和IFN γ的基因表达水平均明显高于pVAON33 S2 S质粒。结论 :C3d P2 8可在提高体液免疫应答的同时增强基因免疫诱导的HBV特异性细胞免疫应答。Objective:To investigate whether P28 derived from C3d can regulate the specific cellular immunoresponse against HBV preS2/S induced by directly injection of naked plasmids DNA containing four tandem repeats of P28 and HBV preS2/S in fusion form. Methods:BALB/C mice were immunized i.m. with 100 μg DNA of pVAON33 S2/S plasmid, pVAON33 S2/S P28 4 plasmid and mock DNA, respectively The levels of specific anti HBs IgG and its isotype distribution in sera collected at the indicated times from each group were determined by ELISA. Splenocytes from immunized mice were stimulated with HBsAg protein and then harvested to analyze specific lympho proliferative response, mRNA level of IL 4 and IFN γ, specific CTL activity by 3H Thymidine incorporation assay, semi quantitative RT PCR, 3H Thymidine release assay, respectively.Results:The lympho proliferative response,levels of HBsAg specific IgG, CTL activity in the mice immunized with pVAON33 S2/S P28 4 was significant higher than that in pVAON33 S2/S immunized mice. IgG1 levels and, unexpectedly, IgG2a and IgG2b levels alike in the mice immunized with pVAON33 S2/S P28 4 were higher than that in the mice with pVAON33 S2/S The significant increase of both IL 4 and IFN γ mRNA levels was detected in mice with S2/S P28 4 expressing DNA compared with mice with S2/S expressing DNA. Conclusion:P28 C3d could enhance cellular immunoresponse against HBV induced by gene immunization, while not influencing the enhancement of antibody responses.

关 键 词：基因免疫 免疫调节 补体C3d 乙型肝炎病毒表面抗原 

分 类 号：R392.12[医药卫生—免疫学]