CD11c细胞的改良法磁珠分离及诱导T细胞增殖研究

Studies of modified magnetic beads sorting of CD11c cells and the role on inducing T cells proliferation

目的:改良CD11c细胞的磁珠分离方法,并观察CD11c细胞诱导同源CD4^+T和CD8^+T细胞增殖的作用。方法:使用胶原酶、DNase I和EDTA处理脾组织,以及小鼠血清和抗CD16/32抗体阻断CD11c磁珠与脾细胞非特异结合后分离CD11c细胞;流式细胞术分析CD11c细胞的纯度;淋巴细胞混合培养和ELISA检测CD11c细胞诱导同源T细胞增殖及分泌细胞因子的作用。结果:改良后的磁珠分离法获得了平均(4.52±0.05)×10~6/鼠的CD11c细胞,纯度高达98%。重组肿瘤疫苗E.coli LLO/ OVA免疫小鼠的CD11c细胞明显促进了CD4^+T和CD8^+T细胞增殖并分泌IL-2和IFN-γ。结论:改良法磁珠分离获得了较多高纯度的CD11c细胞,活化的CD11c细胞具有诱导同源T细胞增殖及分泌细胞因子的作用。

Objective：To modify the manipulation of CD11c cells magnetic beads sorting,and explore the effect of CD11c cells on inducing the proliferation of autogenic CD4＋T and CD8＋T cells. Methods： Collagenase, DNase Ⅰ and EDTA were used for digesting the spleen tissue, and murine serum and CD16/32 antibody were applied for blocking the nonspecific connecting of CD11c beads and the spleen cells. After modified beads sorting, the purity of CD11c cells was checked by flow cytometry. The proliferation of autogenic CD4＋F and CD8T cells and the concentration of cytokines in the supernatant of the cocuhue cells were detected by mixed lymphocyte culture and ELISA,respectively. Results：Average （4.52 ± 0.05）×10^6 CD11c cells were collected in each murine spleen by the modified beads sorting,and the purity of CD11c was 98%. The proliferation of CD4＋T and CD8CF cells and the secretion of IL-2 and IFN-γ of these cells were significantly induced by CD11c cells of recombinant E.coli LLO/OVA vaccinated mice. Conclusion：More quantity and higher purity of CD11c cells were collected by modified magnetic beads sorting, and the actived CD11c cells play a role on inducd autogenic T cells proliferation and cytokines secretion.