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目的探讨黑色素瘤小鼠体内CD8+T淋巴细胞亚群的比例变化并对CD8+CD62L+CD127+T细胞亚群的功能做初步研究。方法将B16细胞注入C57/BL小鼠右腋皮下,制备黑色素瘤小鼠模型,4周后处死,用流式细胞术检测脾脏、血液及骨髓细胞中各CD8+T淋巴细胞亚群的比例变化,将升高的CD8+CD62L+CD127+T细胞群与肿瘤细胞共培养,流式检测其凋亡、ELISA检测共培养上清中IFN-γ的分泌水平;将CD8+CD62L+CD127+T细胞体外刺激后用MTT法检测其增殖情况。结果在B16荷瘤小鼠脾脏CD8+CD62L+、CD8+CD127+和CD8+CD62L+CD127+T细胞频率显著高于正常对照小鼠,外周血和骨髓中这些细胞的频率在2组小鼠之间无显著性差异。荷瘤小鼠体内CD8+CD62L+CD127+T细胞亚群IFN-γ分泌的能力低于CD8+CD62L+CD127-T细胞(P<0.05),也明显低于CD8+T细胞(P<0.01)。但相对于CD8+T细胞,CD8+CD62L+CD127+T增殖能力明显增强(P<0.01)而凋亡减弱(P<0.05)。结论 B16荷瘤小鼠脾脏中记忆CD8+T细胞亚群频率升高,而CD8+CD62L+CD127+T细胞亚群有更强的增殖能力和抗凋亡能力,提示小鼠黑色素瘤促进了记忆CD8+T细胞生成且CD8+CD62L+CD127+T细胞可能具有更独特的作用。
Objective To investigate the proportion of CD8 + T lymphocyte subsets in melanoma mice and to study the function of CD8 + CD62L + CD127 + T cell subsets. Methods B16 cells were injected subcutaneously into the right armpit of C57 / BL mice to prepare a mouse model of melanoma. After 4 weeks, B16 cells were sacrificed. The proportion of CD8 + T lymphocyte subsets in spleen, blood and bone marrow cells was measured by flow cytometry The CD8 + CD62L + CD127 + T cells were co-cultured with tumor cells and the apoptosis was detected by flow cytometry. The secretion of IFN-γ in the co-culture supernatants was detected by ELISA. After in vitro stimulation MTT assay proliferation. Results The frequencies of CD8 + CD62L +, CD8 + CD127 + and CD8 + CD62L + CD127 + T cells in spleen of B16 tumor-bearing mice were significantly higher than those in normal control mice. The frequency of these cells in peripheral blood and bone marrow was no significant difference between the two groups Significant difference. The ability of IFN-γ secretion of CD8 + CD62L + CD127 + T cell subsets in tumor-bearing mice was lower than that of CD8 + CD62L + CD127-T cells and also significantly lower than that of CD8 + T cells (P <0.01) . However, compared with CD8 + T cells, CD8 + CD62L + CD127 + T enhanced proliferation (P <0.01) and attenuated apoptosis (P <0.05). Conclusions The frequency of memory CD8 + T lymphocyte subsets in spleen of B16 tumor-bearing mice is increased, while CD8 + CD62L + CD127 + T cell subsets have stronger proliferative ability and anti-apoptotic ability, suggesting that mouse melanoma promotes memory CD8 + T cells and CD8 + CD62L + CD127 + T cells may have a more unique role.