HPV16 CTL表位E749-57以HSP110为分子伴侣的免疫原性研究

doi:10.3760/cma.j.issn.0412-4030.2010.05.016

科研成果详情

题名	HPV16 CTL表位E749-57以HSP110为分子伴侣的免疫原性研究
其他题名	Immunogenicity of the immunodominant cytotoxic T lymphocyte epitope E749-57 in HPV16 oncoprotein E7 chaperoned by HSP110
作者	任发亮 1,2; 徐云升1,2; 欧荣英1,3; 倪兵 4; 贾正才 4; 吴玉章 4; 林治华 5; 李秉煦1,2; 郝飞 6
发表日期	2010-05-15
发表期刊	中华皮肤科杂志影响因子和分区
语种	中文
原始文献类型	学术期刊
关键词	人乳头状瘤病毒16 表位 T淋巴细胞 HSP110热休克蛋白质类
其他关键词	Human ; papillomavirus 16 ; Epitopes ; T-lymphocytes ; HSP110 heat-shock proteins
摘要	目的探讨以mHSP110为分子伴侣的HPV16 CTL表位E7_(49-57)的免疫原性。方法将mHSP110基因克隆、原核表达和纯化,SDS-PAGE和Western blot鉴定。在热休克状态与E7_(49-57)结合形成复合物,高压液相色谱(HPLC)鉴定其结合程度。用mHSP110-E7_(49-57)复合物免疫小鼠,IFN-γ胞内染色、MTT法检测小鼠脾细胞中特异CTL。结果克隆mHSP110片段经DNA序列测定与基因库中其CDS一致,长度为2577 bp;经SDS-PAGE和Western印迹证实mHSP110表达、纯化成功,HPLC分析E7_(45-57)能够与mHSP110形成复合物。复合物免疫小鼠的脾淋巴细胞中CD8~+IFN-γ~+T细胞的频率、脾淋巴细胞增殖活性明显高于E7_(49-57)组、HSP110组和PBS组。复合物免疫小鼠可明显抑制TC-1肿瘤的生长。结论 mHSP110-E7_(49-57)复合物能诱导产生特异性CTL并产生抗肿瘤效应。
其他摘要	Objective To investigate the immunogenicity of immunodominant cytotoxic T lymphocyte epitope E749-57 of human papilloma virus (HPV) 16 oncoprotein E7 chaperoned by heat shock protein (HSP)110. Methods Mouse HSP110 gene was cloned into prokaryotic expression vector pQE-80L for the expression of HSP110 protein, which was purified using Ni-NTA column. SDS-PAGE and Western-blot were conducted to confirm the purified mHSP110 protein, which was subsequently incubated with E749-57 peptide under heat shock condition, and high-performance liquid chromatography (HPLC) was used to evaluate the binding efficiency of the recombinant protein and E749-57 peptide. Twenty mice were divided into 4 groups to be immunized with mHSP110 protein, E749-57 peptide, mHSP110-E749-57 complex and phosphate buffered saline (PBS),respectively. Two weeks after the last immunization, spleen cells were collected from the immunized mice and divided into 2 parts: one were stimulated by E749-57 peptide followed by the detection of CD8+ INF-γ+ T cells with flow cytometry; the other one were subjected to MTT analysis for the estimation of cell proliferation. The mHSP110-E749-57 complex was also used to immunize TC-1 tumor bearing mice to observe its anti-tumor effect.Results The full-length 2577 bp-sized mHSP110 gene was amplified from mouse liver cDNA and cloned into pQE-80L vector. Direct sequencing confirmed the correctness of the cloning. SDS-PAGE and Western-blot demonstrated the successful purification of mHSP110. HPLC assay showed that the purified mHSP110 protein could bind with E749-57 to form a relatively stable protein complex. The percentage of IFN-γ+ CD8+ T cells in and proliferation index of spleen cells from the complex-immunized mice were statistically higher than those from the other 3 groups of mice. Moreover, the complex could obviously inhibit the growth of TC-1 tumor in mice. Conclusion The mHSP110-E749-57 complex could enhance the generation of specific cytotoxic T lymphocytes and exert anti-tumor effects in mice.
资助项目	国家自然科学基金（30972800、30500537、60873103）
ISSN	0412-4030
卷号	43 期号:05 页码:346-349
DOI	10.3760/cma.j.issn.0412-4030.2010.05.016
页数	0
收录类别	CNKI ; 万方 ; 北大核心 ; PKU ; CSCD ; ISTIC
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引用统计
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/71558
专题	第一临床医学院（信息与工程学院）、附属第一医院_皮肤病与性病学_皮肤性病科附属第一医院附属第一医院_皮肤科
作者单位	1.温州医学院皮肤性病学研究所; 2.温州医学院附属第一医院皮肤科; 3.温州医学院附属第一医院妇产科; 4.第三军医大学全军免疫学研究所; 5.重庆工学院生物工程学院; 6.第三军医大学西南医院皮肤科
第一作者单位	第一临床医学院（信息与工程学院）、附属第一医院_皮肤病与性病学_皮肤性病科; 附属第一医院_皮肤科
第一作者的第一单位	第一临床医学院（信息与工程学院）、附属第一医院_皮肤病与性病学_皮肤性病科
推荐引用方式 GB/T 7714	任发亮,徐云升,欧荣英,等. HPV16 CTL表位E749-57以HSP110为分子伴侣的免疫原性研究[J]. 中华皮肤科杂志,2010,43(05):346-349.
APA	任发亮., 徐云升., 欧荣英., 倪兵., 贾正才., ... & 郝飞. (2010). HPV16 CTL表位E749-57以HSP110为分子伴侣的免疫原性研究. 中华皮肤科杂志, 43(05), 346-349.
MLA	任发亮,et al."HPV16 CTL表位E749-57以HSP110为分子伴侣的免疫原性研究".中华皮肤科杂志 43.05(2010):346-349.