HPV L1C-末端保守序列多肽免疫学特性的探讨  

To investigate the immunity characteristics of a conserved sequence on the C terminal region of the human papillomavirus major protein L1

作　　者：蒋雪梅[1] 肖长义[1] 吴江峰[1] 李红军[1] 袁太宁[1] 汤桂成[1] 

机构地区：[1]三峡大学医学院,湖北宜昌443002

出　　处：《重庆医学》2014年第1期12-15,共4页Chongqing medicine

基　　金：国家自然科学基金资助项目(30770104);湖北省教育厅项目(B20121306);三峡大学人才科研启动基金项目(KJ2010B045)

摘　　要：目的探讨一段位于人乳头状瘤病毒(HPV)L1C-末端、长30个氨基酸残基的保守序列的免疫学性质而进行此实验。方法以此序列为基础人工合成多肽加氢氧化铝佐剂免疫小鼠。取免疫鼠脾脏制备脾细胞悬液,流式细胞术检测小鼠脾脏中CD3+、CD3+CD4+、CD3+CD8+淋巴细胞数量,计算CD4+/CD8+值。四甲基偶氮唑盐(MTT)法检测细胞增殖。取培养上清液以双抗体夹心法ELISA检测细胞因子IL-4、IFN-γ的含量。结果 (1)脾细胞CD3+CD4+细胞数量、CD4+/CD8+值比阴性对照组明显升高,差异有统计学意义(P<0.05);(2)T细胞增殖实验结果未见明显差异(P>0.05);(3)细胞因子IL-4含量较对照组显著增高,差异有统计学意义(P<0.01)。IFN-γ含量与对照组比较无明显差异(P>0.05)。结论结果表明多肽诱导体液免疫效果是肯定的,有作为预防性疫苗进一步研究开发的前景;多肽的细胞免疫原性可能较弱,未能诱导出与细胞免疫相关的反应,多肽的免疫原性还有待于进一步改进提高。Objective To investigate and detect the immunity characteristics of a conserved sequence including 30 amino residues which is located in the C terminal of HPVL1 .Methods The immune model of mouse was establish with a polypeptide synthetized based on this sequence .The amount of CD3＋ ,CD3＋CD4＋ or CD3＋CD8＋ lymphocyte of the mouse spleen was detected by Flow cy-tometry ,then the value of CD4＋ /CD8＋ were calculated .The cell proliferation was detected by MTT assay .Sample was took from the cell supernatant to detect the content of IL-4 and IFN-γby double antibody sandwich ELISA .Results （1）The amount of CD3＋CD8＋ lymphocytes and the value of CD4＋ /CD8＋ were significantly increased than control group（P〈0 .05） .（2）The result of T cell proliferation showed no significant difference （P〉0 .05） .（3）The level of IL 4 were significantly higher than that of control group （P〈0 .01） .The content of IFN-γlevel showed no significant difference compared with control group （P〉0 .05） .Conclusion The results showed that it was affirmative that the polypeptides induced humoral immunity ,which was worth to be furtherly studied as a preventive vaccine .But its cell immunogenicity is weak ,and the attenmpt to induce the response related to the cell immunogenicity was failed ,and the immunogenicity of the peptide remains to be improved .

关 键 词：HPV 晚期蛋白 保守序列 免疫原性 

分 类 号：R392[医药卫生—免疫学]