为了了解γ-线照射是否影响体外培养的树突状细胞的表型与功能,利用含rhGM-CSF(800 U/ml)和rhIL-4(500U/ml)的RPMI 1640培养基从多发性硬化症病人的外周血单个核细胞(PBMNC)中诱生树突状细胞(DC).在培养的第6天加入5 μg/ml的脂多糖继续培养24小时促使DC完全成熟.于第7天收获DC并等分成几部分,一部分未经γ-线照射的DC用作对照组,其他部分的DC分别用25 Gy和30 Gy剂量的γ射线照射.流式细胞术分析DC的表面分子并测定DC细胞刺激同源T细胞增殖的能力.结果表明,γ-线照射减少树突状细胞CD86,CD80和HLA-DR表达,尤其是CD86分子的表达(P=0.0072).人DC能有效地刺激同源T细胞的增殖,但是同未经照射的DC相比,照射的DC刺激T细胞增殖的能力显著降低.结论:γ-线照射不仅影响DC的表型,而且影响它的功能.%To determine whether gamma irradiation influences phenotype and function of human dendritic cells (DC) in vitro,dendritic cells were induced from the peripheral blood mononuclear cells of multiple sclerosis patients with RPMI 1640 mediumcontaining recombinant human GM-CSF (rhGM-CSF, 800 Ul) and recombinant human IL-4 (rhIL-4, 500 U/ml).Phenotypic changes were monitored by light microscopy. Lipopolysaccharide at a concentration of 5 ug/ml was added into thecultures after 6 days of growth for DC complete maturation, and the cells were cultured for another 24 hours. The harvested DCon day 7 were divided equally into several parts. One part was used as non-irradiated DC (naive DC) while the other parts wereirradiated by γ ray at a dose of 25 Gy and 30 Gy respectively. Cell surface molecules were analyzed by flow cytometry. Thecapability of DC to stimulated autologous T cell proliferation were determined. The results showed that gamma irradiation reducedexpression of CD86, CD80 and HLA-DR molecules on dendritic cells, especially CD86 molecules. Dendritic cells effectivelystimulated autologous T cells proliferation while irradiated DC in all groups showed profound decrcase of capability to promote Tcells proliferation. It is concluded that gamma irradiation of dendritic cells not only influenced phenotype of DC but also alteredtheir function as stimulator cells in mixed lymphocyte reaction.
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