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目的 :研究丙型肝炎病毒 (HCV)核心 (C)基因免疫诱生特异性免疫应答的可行性。方法 :将HCVC基因片段插入真核表达载体pcDNA3质粒CMV启动子的下游 ,构建真核表达载体pcDNAHCV C ,分别转染小鼠骨髓瘤细胞SP2 0和人肝癌细胞 772 1进行瞬时表达 ,用免疫荧光法和Western blot检测表达产物 ;将重组质粒注射BALB c(H 2 d)小鼠股四头肌 ,ELISA法检测血清中抗体产生水平。结果 :2 4只免疫小鼠 ,初次免疫 2w后 ,血清中均出现了HCVC抗体 ,随着免疫次数和免疫剂量的增加 ,抗体滴度明显提高。结论 :HCVC基因免疫能够诱导机体产生特异性的体液免疫应答 ,从而可能进一步诱生细胞免疫应答 ,成为防治HCV感染的有效方法
Objective: To investigate the feasibility of inducing specific immune response induced by the gene of hepatitis C virus (HCV) core (C). Methods: The HCVC gene fragment was inserted into the downstream of the CMV promoter of the eukaryotic expression vector pcDNA3 plasmid. The eukaryotic expression vector pcDNAHCV C was constructed and transfected into mouse myeloma cells SP2 0 and human hepatocellular carcinoma cells 772 1 respectively for transient expression. Immunofluorescence The recombinant plasmids were injected into the quadriceps femoris of BALB / c (H 2 d) mice and the level of antibody in serum was detected by ELISA. RESULTS: Twenty-four immunized mice were immunized with HCVC antibodies in their serum after 2 weeks of initial immunization. The antibody titer was significantly increased with the increase in the number of immunizations and immunization doses. Conclusion: HCVC gene immunization can induce the body to produce specific humoral immune response, which may further induce cellular immune response and become an effective method to prevent and treat HCV infection