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目的比较两种抗原负载方式对树突状细胞(DCs)疫苗的影响。方法抽取HLA表型为A11的健康志愿者外周血,分离单核细胞,体外培养。通过反复冻融LoVo细胞,提取肿瘤细胞裂解物或者使用含CEA片断的重组腺相关病毒转染未成熟DCs,诱导特异性细胞毒性T细胞。检测体外培养的DCs和CTL活性,并使用MTT法检测两组CTL对LoVo细胞的杀伤作用。结果两种方式均可培养的成熟DCs,诱导的CTL细胞分泌IFN-γ有所增加;转染后DCs诱导特异性CTL可有效识别并杀伤HLA-A11阳性的LoVo细胞,腺相关病毒提呈抗原制备的DCs疫苗对LoVo细胞的杀伤作用明显高于肿瘤细胞裂解物的抗原负载方式。结论两种抗原提呈方式均可培养出成熟的DCs,不明显改变DCs表型和刺激淋巴细胞增殖、分化功能,并可诱导自体CTL增殖。使用腺相关病毒转染DCs的方式明显优于肿瘤细胞裂解物的抗原负载方式。
Objective To compare the effects of two antigen loading methods on dendritic cell (DCs) vaccines. Methods Peripheral blood from healthy volunteers with HLA phenotype A11 was collected and monocytes were isolated and cultured in vitro. Specific cytotoxic T cells are induced by repeated freezing and thawing of LoVo cells, extraction of tumor cell lysates, or transfection of immature DCs with recombinant adeno-associated virus containing the CEA fragment. The activity of DCs and CTLs in vitro were detected. MTT assay was used to detect the killing effect of CTL on LoVo cells. Results The mature DCs cultured in both ways increased the secretion of IFN-γ by CTL cells. After transfection, the specific CTL induced by DCs could effectively identify and kill HLA-A11-positive LoVo cells. The adeno-associated virus presented antigens The killing effect of DCs vaccine on LoVo cells was significantly higher than that of tumor cell lysate. Conclusion Both methods of antigen presentation can produce mature DCs without changing the phenotype of DCs and stimulating the proliferation and differentiation of lymphocytes and inducing the proliferation of autologous CTL. The use of adeno-associated virus to transfect DCs significantly outperforms the antigen loading of tumor cell lysates.