论文部分内容阅读
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 improve the method of magnetic beads separation of CD11c cells and to observe the effect of CD11c cells on the proliferation of homologous CD4 ~ + T and CD8 ~ + T cells. Methods: The spleen tissues were treated with collagenase, DNase I and EDTA, and CD11c cells were isolated after non-specific binding of CD11c magnetic beads and spleen cells with mouse serum and anti-CD16 / 32 antibody. Flow cytometry was used to analyze the purity of CD11c cells. Lymphocyte mixed culture and ELISA assay CD11c cells induced homologous T cell proliferation and secretion of cytokines. Results: The average percentage of CD11c cells (4.52 ± 0.05) × 10 ~ 6 / mouse was obtained by the improved magnetic bead separation method with purity of 98%. The CD11c cells of mice immunized with recombinant tumor vaccine E.coli LLO / OVA significantly enhanced the proliferation of CD4 + T and CD8 + T cells and secreted IL-2 and IFN-γ. Conclusion: The CD11c cells with high purity were obtained by the improved magnetic beads separation. The activated CD11c cells induced the proliferation and secreted cytokines of the homologous T cells.