小鼠肝素酶H-2Kb限制性CTL表位的实验鉴定

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。

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

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 B16 melanoma cell(mHpa+,H-2Kb+)、Lewis lung carcinoma cell(mHpa+,H-2Kb+)、EL-4 leukoma cells(mHpa+,H-2Kb+) and P815 mastocytoma cells(mHpa+,H-2Kd+) will be measured by standard 51Cr release test respectively.IFN-γ released by effectors was measured by ELISPOT method.Result Of five peptides tested,only peptide mHpa398-405 and mHpa519-526 were able to elicit mHpa-specific CTLs,which could lyse Lewis lung carcinoma cells,B16 melanoma cells and EL-4 lymphoma cells expressing both mHpa and H-2Kb.But they can not lyse P815 mastocytoma cells(mHpa+,H-2Kb-).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.