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目的观察IκBα突变体基因修饰的树突状细胞(IκBαM-DC)对同种T细胞的反应性。方法利用腺病毒载体将IκBαM基因转染WF大鼠骨髓树突状细胞(DC),Western-blot法检测DC中IκBα、IκBαM基因的表达;用流式细胞仪检测DC中共刺激分子MHCⅡ、CD80、CD86、CD40的表达;酶联免疫法(ELISA)分析DC分泌IL-12的含量。通过混合淋巴细胞反应(MLR)分析Lewis大鼠T细胞对IκBαM-DC刺激的增殖能力,二次MLR检测IκBαM-DC诱导的T细胞抗原特异性的低反应性。结果IκBαM抑制DC共刺激分子MHCⅡ、CD80、CD86、CD40的表达及IL-12分泌。同种T细胞对IκBαM-DC刺激的增殖能力较未转染的DC反应明显降低;T细胞低反应具有抗原特异性。结论表达IκBα突变体基因的DC能降低同种T细胞的反应性。
Objective To observe the reactivity of IκBα gene-modified dendritic cells (IκBαM-DC) to allogeneic T cells. Methods The IκBαM gene was transfected into bone marrow dendritic cells (DCs) of WF rats by adenoviral vector. The expression of IκBα and IκBαM genes in DCs was detected by Western-blot. The expressions of co-stimulatory molecules MHCⅡ, CD80, CD86, CD40 expression; ELISA analysis of DC secretion of IL-12 content. The proliferation of Lewis rat T cells stimulated by IκBαM-DC was analyzed by mixed lymphocyte reaction (MLR). The secondary MLR was used to detect IκBαM-DC-induced T cell antigen-specific hyporesponsiveness. Results IκBαM inhibited the expression of DC costimulatory molecules MHCⅡ, CD80, CD86, CD40 and IL-12 secretion. Allogeneic T cells on IκBαM-DC stimulated proliferation than non-transfected DC response was significantly lower; T cell low response antigen-specific. Conclusion DC expressing IκBα mutant gene can reduce the reactivity of allogeneic T cells.