丙型肝炎病毒重组蛋白抗原联合免疫的免疫原性和保护性研究  

Immunogenicity and protective effect induced by co-immunization with HCV recombinant protein antigens

作　　者：曾瑞红[1] 李广学[1,2] 凌世淦[3] 张贺秋[3] 姚智燕[1] 杨建岭[1] 贺峰[3] 黄睿[3] 刘艳坤[4] 魏林[1] 

机构地区：[1]河北医科大学免疫教研室,石家庄050017 [2]邢台医学院 [3]军事医学科学院基础医学研究所 [4]河北医科大学生物工程中心

出　　处：《中华微生物学和免疫学杂志》2009年第7期642-645,共4页Chinese Journal of Microbiology and Immunology

基　　金：河北省科技支撑计划项目（08276412D）

摘　　要：目的研究两个丙型肝炎病毒（HCV）多表位重组抗原联合免疫后诱导的细胞免疫和体液免疫应答，以及对小鼠的保护作用。方法用两个多表位抗原HCV-T和HCV—E1联合免疫BALB／c小鼠3次，ELISA检测血清中抗体IgG、IgG1和IgG2a滴度；最后一次免疫后10d杀死一半小鼠，取脾细胞，用乳酸脱氢酶（LDH）释放法检测细胞毒性T淋巴细胞（CTL）活性；EUSPOT法检测分泌IFN-γ和IL-4的细胞；最后一次免疫后2周，在免疫小鼠的背部皮下注射10^6个SP2／0-NS3细胞，考察联合免疫对小鼠的保护作用；另取BALB／c小鼠，先在背部皮下注射10^6个SP2／0-NS3细胞，7d后开始联合免疫，免疫3次，考察免疫治疗作用。结果用HCV—T＋HCV—E1联合免疫诱导了高滴度的HCV-E1特异性的IgG、IgG1和IgG2a抗体以及高水平的CTL活性；与单独免疫相比，联合免疫产生的分泌IFN-γ和IL-4的细胞数量有协同作用；而且联合免疫能有效预防和治疗SP2／0-NS3对小鼠的攻击。结论HCV—T＋HCV—E1联合免疫诱导了保护性的体液免疫和细胞免疫应答，有望进一步开发为一种有效的重组HCV疫苗。Objective To investigate the cellular and humoral immune responses and protective effect induced by co-immunization with two muhi-epitope combinant antigens. Methods Mice were co-immunized with the multi-epitope HCV-T and HCV-E1 antigens three times. Sera antibodies IgG, IgG1 and IgG2a were tested by ELISA. Spleens from BALB/c mice immunized were removed 10 days after the last immunization. CTL activity was assessed using LDH cytotoxicity assay kit. IFN-γ- and IL-4-secreting cells were quantified using ELISPOT kit. Two weeks after the final immunization, the mice were challenged subcutaneously（ s. c. ） at the back with 10^6 SP2/0-NS3 cells, and protective effect was observed. For therapy, 10^6 SP2/0-NS3 cells were implanted into the back of BALB/c mice. Seven days later, mice were immunization three times. Therapy effect was observed. Results Co-immunization with HCV-T and HCV-E1 induced high tiers of HCV-E1-specific IgG, IgG1 and IgG2a antibodies, and high level of CTL activity. Synergistic effect in frequencies of both specific IFN-γ-secreting cells and IL-4-secreting cells was observed in mice coimmunized. Prophylactic as well as therapeutic administration of mT ＋ mE1 in mice led to protecting mice against SP2/0-NS3 ceils. These results suggested that mT ＋ mE1 was potential as a prophylactic as well as therapeutic HCV vaccine. Conclusion Co-immunization with HCV-T ＋ HCV-E1 induced protective humoral and cellular immune response. HCV-T ＋ HCV-E1 was potential as a recombinant HCV vaccine.

关 键 词：丙型肝炎病毒 多表位抗原 联合免疫 免疫原性 

分 类 号：R3[医药卫生—基础医学]