地方株HPV16L1基因诱发的小鼠体液免疫反应  

L1 Gene of Local Strain HPV16 Induces Immuno-humoral Response in Mice

作　　者：高慧[1] 孙强 黄正芳 吴宝金[1] 李厚达 

机构地区：[1]扬州大学医学院,扬州225001 [2]扬州大学比较医学中心,扬州225009

出　　处：《实验动物与比较医学》2005年第3期143-146,共4页Laboratory Animal and Comparative Medicine

基　　金：江苏省自然科学基金基础研究项目资助(BK2004057)

摘　　要：目的检测pcDNA-L1的表达,评价地方株HPV16L1基因疫苗诱导的体液免疫反应。方法采用PCR技术从宫颈癌组织中获得L1基因,以pGEM-T Easy为克隆载体构建重组质粒 pGEM-T-L1,进行限制性核酸内切酶及核苷酸序列分析,再将L1基因亚克隆到真核表达载体 pcDNA3．1(-)中,构建地方株HPV16L1基因疫苗,转染COS-7细胞,通过IFA试验验证L1蛋白的表达。用pcDNA-L1肌肉内注射方法分别于第1天、第15天和第43天免疫6周龄BABL/c小鼠, 在特定时间段采血分离血清,IFA分析基因免疫在小鼠体内诱导的免疫应答,设pcDNA载体对照。结果 pcDNA-L1能够在COS-7细胞中暂态表达,免疫小鼠后能检测出特异的抗体,用IFA方法在第一次免疫后的第30、60和180天均可检测到抗体;原载体免疫后未检出特异抗体。结论本实验构建的地方株HPV16L1基因疫苗可诱导产生特异性的体液免疫反应。Objective To test the expression of recombinant plasmid pcDNA-L1 and evaluate its efficiency to induce specific humoral immune responses. Methods The local HPV16-L1 gene was generated by polymerase chain reaction, inserted into pGEM-T Easy cloning vector, and then subcloned into the eukaryotic expression vector pcDNA3.1（-）. The expression of recombinant plasmid pcDNA-L1, transferred in Cos-7 cells, was identified by immunofluorescence. The 6-week-old BALB/c m/ce were immunized via muscular 3 times with pcDNA-L1, thereafter the sera antibody of were measured by immunofluorescence and western-blot. Results With the immunofluorescence technique, L1 protein was found to express transiently in the transfected cells of the pcDNA-L1. Specific anti-HPV16-L1 antibodies could be detected by IFA at the end of 30,60 and 180 days after the first immunization, No specific antibodies could be detected in the control group of pcDNA3.1 （-）. Conclusions The recombinant plasmid pcDNA-L1 DNA constructed by us could induce specific immuno-humoral response in mice.

关 键 词：HPV16 L1基因 体液免疫 小鼠 

分 类 号：R392[医药卫生—免疫学]