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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 experimentally identify H-2Kb-restricted mouse heparinase (mHpa) CTL epitopes. Methods Five mHpa epitopes were loaded on bone marrow-derived dendritic cells of C57BL / 6 mice respectively and induced to generate epitope-specific effector cells. The standard 4 h51Cr release assay was used to detect the effect of effector cells on different target cells. ELISPOT was used to detect the ability of effector cells to secrete IFN-γ. Results Of the five mHpa CTL epitopes, only mHpa398-405 and mHpa519-526 induced specific CTL production in mice, which showed significant immunogenicity against B16 melanoma cells, Lewis lung carcinoma cells and EL-4 lymphoma cells Effect, and H-2Kb-negative P815 mastocytoma cells do not have immune-killing effect, further study found that the peptide-specific CTL on autologous lymphocytes and DC cells have no killing effect, on the other hand, mHpa398-405 and mHpa519 -526 also promotes the ability of effector cells to secrete IFN-γ. Conclusion mH-pa398-405 and mHpa519-526 are mouse heparinase-derived and restricted by H-2Kb CTL epitopes. Mice can induce epitope-specific CTL responses in vivo.