论文部分内容阅读
目的通过树状串联hTERT表位肽的髓样树突状细胞(mDC)递呈,刺激同源淋巴细胞,探索一种更优的肿瘤免疫治疗方法。方法人工固相合成4分支的树状串联hTERT表位肽(MAP)及其各分支单肽,免疫荧光检测hTERT的表达情况,免疫磁珠分选mDC,尼龙毛柱纯化T细胞,ELISA检测mDC的IL-12p70和淋巴细胞的(LC)TNF-α、IFN-γ分泌量,流式细胞技术检测mDC、LC的相关表面分子以及效应性T细胞对HLA-A2型肿瘤A549、MDA-MB-231和SW480的杀伤率,并作统计学分析。结果所有肿瘤细胞都表达hTERT,且胞核大于胞浆;MAP和混合单肽对3种肿瘤细胞都有杀伤效应,且MAP的杀伤效应大于混合单肽,有统计学差异。结论人工合成hTERT的MAP肽通过mDC能足够强的激活同源淋巴细胞,在肿瘤疫苗的研发中有重要意义。
OBJECTIVE: To explore a more effective method of tumor immunotherapy by presenting dendritic cells dendritic cells (mDC) with dendritic-in-tandem hTERT epitope peptides to stimulate homologous lymphocytes. Methods The 4-branched dendritic hTERT epitope peptide (MAP) and its branched single peptides were synthesized by artificial solid-phase method. The expression of hTERT was detected by immunofluorescence. The mDC was sorted by immunomagnetic beads and the T cells were purified by nylon column. The expression of TNF-α and IFN-γ in IL-12p70 and lymphocyte (LC) were detected by flow cytometry. The effect of related surface molecules of mDC, LC and effector T cells on the expression of HLA- 231 and SW480 killing rate, and for statistical analysis. Results All the tumor cells expressed hTERT, and the nucleus was larger than the cytoplasm. Both MAP and mixed monospecifics had killing effect on all three kinds of tumor cells, and the killing effect of MAP was greater than that of mixed single peptide with statistical significance. CONCLUSION: MAP peptide synthesized by hTERT can strongly activate homologous lymphocytes through mDC, which is of great significance in the development of tumor vaccines.