摘要目的 探讨热休克蛋白(HSP)110对人乳头瘤病毒(HPV)16型E711-20配体的免疫佐剂效应.方法 体外构建HSP 110与HPV 16型E711-20配体的复合物.将C57BL/6雌性6周龄小鼠15只随机分为3组:复合物组、配体组和磷酸盐缓冲液(PBS)组,每组5只,腹腔注射免疫复合物100 μg,肽10μg,PBS100μl,2周后重复1次,第2次免疫2周后,分离脾细胞作为效应细胞.免疫小鼠后采用噻唑蓝法(MTT)判断脾淋巴细胞增殖活性,干扰素γ(IFN-γ)胞内染色法检测小鼠脾细胞中特异性细胞毒T淋巴细胞,并以标准51Cr释放实验检测特异性细胞毒T淋巴细胞对靶细胞的杀伤效应.采用SPSS10.0软件进行t检验.结果 HSP110与HPV16型E711-20配体的复合物免疫小鼠后脾淋巴细胞增殖活性(增殖指数为1.87±0.12)和CD8+ IFN-γT细胞的频率(3.9％)显著高于配体组(分别为0.32±0.071和0.4％),差异均有统计学意义(P＜0.01).配体的复合物对靶细胞的杀伤效应(效靶比为100∶1、50∶1、25∶1、12.5∶1时,杀伤率分别为54.7％、72.2％、61.5％、39.8％)显著高于配体组(分别为35.2％、49.3％、28.1％、17.4％).结论 HSP110能增强HPV16型711-20配体的免疫效应,具有较好的免疫佐剂作用.

abstractsObjective To investigate the adjuvant effect of heatshock protein 110 (HSP110) on the immune responses induced by an altered peptide ligand of human papilloma virus type 16 E711-20 peptide (HPV16E711-20).Methods The complex of HSP110 and an altered peptide ligand of HPV16E711-20 was constructed in vitro.Fifteen 6-week-old C57BL/6 female mice were randonly and equally divided into 3 groups,including complex group,ligand group,and phosphate buffered solution (PBS) group,to receive intraperitoneal immunization with the complex (100 μg),peptide (10 μg),and PBS (100 μl) respectively.Immunization was carried out with an interval of 2 weeks for 2 times.Two weeks after the last immunization,the mice were sacrificed followed by the isolation of splenocytes.MTT assay was performed to evaluate the proliferation activity of splenocytes,intracellular staining for interferon (INF)-γ to detect cytotoxic T lymphocytes (CTLs),standard chromium-51 (51Cr) release assay to estimate the lethal effect of specific CTLs on target cells.Statistical analysis was performed by using t test with SPSS 10.0 software.Differences were considered as statistically significant when the P value was less than 0.05.Results A significant increase was observed in the proliferation index ( 1.87 ± 0.122 vs.0.32 ± 0.071,t =4.01,P ＜ 0.01 ) of,and percentage of CD8+IFN-γ+ T lymphocytes (3.9％ vs.0.4％,t =3.88,P ＜ 0.01 ) among splenocytes from the complex group compared with the ligand group.At the effector-to-target ratio of 100 ∶ 1,50 ∶ 1,25∶ 1 and 12.5 ∶ 1,the death rate of target cells was 54.7％,72.2％,61.5％ and 39.8％ respectively after incubation with CTLs from the compleximmunized mice,higher than that from the ligand-immunized mice (35.2％,49.3％,28.1％,17.4％,respectively).Conclusion HSP110 could enhance the immunological effect of the altered peptide ligand of HPV16E711-20,and can serve as an immunological adjuvant.