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热休克蛋白110对人乳头瘤病毒16型E711-20配体的佐剂效应研究
引用本文:徐云升,欧荣英,张学奇,李智铭,李秉煦. 热休克蛋白110对人乳头瘤病毒16型E711-20配体的佐剂效应研究[J]. 中华皮肤科杂志, 2012, 45(4): 266-269
作者姓名:徐云升  欧荣英  张学奇  李智铭  李秉煦
作者单位:1. 温州医学院附属第一医院2. 浙江温州医学院附属第一医院妇产科3. 温州医学院第一医院皮肤性病科
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10 μg,PBS 100 μl,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-γ+ T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。

关 键 词:修饰多肽配体  
收稿时间:2011-11-16

Heat shock protein 110 improves the immunological effect of an altered peptide ligand of human papilloma virus type 16 E711-20 peptide
XU Yun-sheng , OU Rong-ying , ZHANG Xue-qi , LI Zhi-ming , LI Bing-xu. Heat shock protein 110 improves the immunological effect of an altered peptide ligand of human papilloma virus type 16 E711-20 peptide[J]. Chinese Journal of Dermatology, 2012, 45(4): 266-269
Authors:XU Yun-sheng    OU Rong-ying    ZHANG Xue-qi    LI Zhi-ming    LI Bing-xu
Abstract:Objective 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 randomly 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 complex-immunized 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.
Keywords:HSP110 heat-shock proteins  Human papillomavirus 16  Altered peptide ligand
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