乳腺癌相关抗原ROR1 H-2Kd限制性CTL表位预测及初步鉴定  

Prediction and identification of H-2Kd restricted CTL epitope derived from breast cancer associated antigen ROR1

作　　者：马天明[1] 张娜[1] 郑维萍[1] 陈黎[1] 唐俐[1] 苏新良[2] 

机构地区：[1]重庆医科大学基础医学院病理生理学教研室组织工程与干细胞研究室 [2]重庆医科大学附属第一医院内分泌乳腺外科,重庆400016

出　　处：《重庆医科大学学报》2016年第2期168-171,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金面上资助项目(编号:NSFC81272571);重庆市渝中区科技计划资助项目(编号:20120208);重庆市渝中区科技计划资助项目(编号:20110309)

摘　　要：目的:预测并鉴定乳腺癌相关抗原受体酪氨酸激酶样孤儿受体1(receptor tyrosine kinase like orphan receptor 1,ROR1)H-2Kd限制性细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)表位,为研究以ROR1为靶点的乳腺癌免疫治疗奠定基础。方法:采用生物信息学方法预测乳腺癌相关抗原ROR1 H-2Kd限制性CTL表位,选取预测软件中评分较高的抗原表位肽进行人工合成及纯化。肽结合试验检测抗原表位肽与H-2Kd分子的亲和力。选用亲和力较高的抗原表位肽体外刺激脾淋巴细胞,ELISA法检测细胞因子γ-干扰素(interferon-γ,IFN-γ)和白介素-12(interleukin-12,IL-12)的分泌。结果:预测的4条候选ROR1 CTL表位肽均具有较高的亲和力,其荧光指数(fluorescence index,FI)在30μg/ml时均大于1.5。ELISA结果显示表位肽ROR1(NYMFPSQGI)可在体外有效诱导IFN-γ和IL-12细胞因子的产生。结论:ROR1(NYMFPSQGI)为乳腺癌相关抗原ROR1H-2Kd限制性CTL表位,为研究以ROR1为靶点的乳腺癌免疫治疗奠定了基础。Objective：To predict and identify breast cancer associated antigen receptor tyrosine kinase like orphan receptor 1（ROR1）H-2Kd restricted cytotoxic T lymphocyte（CTL）epitopes and to provide experimental data for breast cancer immunotherapy based on ROR1. Methods：ROR1 specific H-2Kd restricted CTL epitopes were predicted by bio-informatic methods. Four candidate epitopes with relatively high scores were artificially synthesized and purified. Analysis of binding ability of the candidate epitope peptides with H-2Kd molecules was carried out by peptide binding assay. ELISA assay was used to check interferon-γ（IFN-γ）and interleukin-12（IL-12）. Results：Four candidate epitopes had high affinities of H-2Kd;the FI values were more than 1.5 when concentrations were more than 30 μg/ml. ELISA results showed that ROR1（NYMFPSQGI）could stimulate spleen lymphocyte secreting IFN-γ and IL-12.Conclusion：ROR1（NYMFPSQGI）is a H-2Kd restricted CTL epitope of breast cancer associated antigen ROR1. This study provides an experimental data for breast cancer immunotherapy based on ROR1.

关 键 词：乳腺癌 ROR1 CTL表位 生物信息学方法 

分 类 号：R730.51[医药卫生—肿瘤]