小鼠肝素酶H-2Kb限制性CTL表位的实验鉴定  

Identification of H-2Kb-restricted CTL epitopes derived from Heparanase antigen

作　　者：汤旭东[1] 万瑛[2] 房殿春[1] 陈陵[1] 熊震[1] 余松涛[1] 师红磊[1] 郭军[1] 杨仕明[1] 

机构地区：[1]第三军医大学西南医院全军消化病研究所,重庆400038 [2]第三军医大学基础医学部全军免疫研究所,重庆400038

出　　处：《实用临床医药杂志》2008年第1期5-11,共7页Journal of Clinical Medicine in Practice

基　　金：国家自然科学基金资助项目(30570841)

摘　　要：目的实验鉴定H-2Kb限制性小鼠肝素酶(mHpa)CTL表位。方法5条mHpa表位分别负载C57BL/6小鼠骨髓来源树突状细胞,并进一步诱导产生表位特异性的效应细胞,采用标准4 h51Cr释放试验检测效应细胞对不同靶细胞的免疫杀伤效应;采用ELISPOT检测效应细胞分泌IFN-γ的能力。结果5条mHpa CTL表位中,仅mHpa398-405和mHpa519-526可以诱导小鼠产生特异性CTL,对同来源的B16黑色素瘤细胞、Lewis肺癌细胞和EL-4淋巴瘤细胞具有明显的免疫杀伤效应,而对H-2Kb阴性的P815肥大细胞瘤细胞不具有免疫杀伤效应,进一步研究发现,上述多肽特异性CTL对自体淋巴细胞和DC细胞不具有杀伤效应,另一方面,mHpa398-405和mHpa519-526还可促进效应细胞分泌IFN-γ的能力。结论mH-pa398-405、mHpa519-526为小鼠肝素酶来源且受H-2Kb限制性CTL表位,小鼠体内可以诱导产生表位特异性的CTL反应。Objective To identify the H-2Kb-restricted epitopes derived from murine Heparanase. Method The specific lysis of mHpa-specific CTL induced by mHpa epitopes on BI6 melanoma cell （mHpa^＋ , H-2K^b＋ ） ,Lewis lung carcinoma cell （mHpa^＋ , H-2K^b＋ ） ,EL-4 leukoma cells （mHpa^＋ , H-2K^b＋ ） and 13815 mastocytoma cells （mHpa＋ , H-2Ke＋ ） will be measured by standard Cr release test respectively. IFN-γ released by effectors was measured by ELISPOT method. Result Of five peptides tested, only peptide mHpa398-405 and mHpaS19-526 were able to elicit mHpa-specific CTLs, which could lyse Lewis lung carcinoma cells, B16 melanoma cells and EL-4 mymphoma cells expressing both mHpa and H-2Kb. But they can riot lyse/3815 mastocytoma cells （mHpa^＋ , H-2K^b- ）. Further research shows mHpa epitopes can induce IFN-γ release in effectors. Conclusion Two H-2Kb-restricted CTL epitopes derived from murine Heparanse, mHpa398-405 （LSLLFKKL） and mHpa519-526 （FSYGFFVI）, were determined for the first time.

关 键 词：肝素酶 H-2Kb限制性 抗原表位 肿瘤疫苗 细胞毒T淋巴细胞 

分 类 号：R730.5[医药卫生—肿瘤]