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目的探讨转染人细胞毒性T淋巴细胞相关抗原4免疫球蛋白(CTLA4Ig)基因的树突状细胞(DCsRev)对反应性T细胞的作用。方法通过重组逆转录病毒将目的基因CTLA4Ig转染到大鼠骨髓来源的树突状细胞(DCs)中。采用逆转录聚合酶链反应(RT-PCR)和斑点酶联免疫吸附试验(Dot-ELISA)检测CTLA4Ig在DCs中的表达;采用混合淋巴细胞反应(MLR)检测DCsRev对反应性T细胞的作用。结果检测结果证实CTLA4Ig基因成功转染至DCs。DCsRev的数量以及用DCsRev预处理反应性T细胞的时间长短与抑制MLR中T细胞增殖有一定的效应关系。当DCs-Rev数量在103~104之间时,其抑制率最高,抑制率可达到为69.12%;用DCsRev预处理反应性T细胞12h时,其对MLR中T细胞增殖的抑制率最高,为98.3%,12h以后,随着预处理时间的延长,抑制率却不断下降。经DCsRev诱导的大鼠体内脾淋巴细胞在MLR中增殖低下。结论转染CT-LA4Ig基因的DCs不但丧失了刺激MLR的能力,并且能够抑制MLR中反应性T细胞的增殖,提示DCsRev可能诱导抗原特异性T细胞的免疫耐受。
Objective To investigate the effect of dendritic cells (DCsRev) transfected with human cytotoxic T lymphocyte associated antigen 4 immunoglobulin (CTLA4Ig) gene on reactive T cells. Methods The target gene CTLA4Ig was transfected into rat bone marrow-derived dendritic cells (DCs) by recombinant retrovirus. The expression of CTLA4Ig in DCs was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Dot-ELISA. The mixed lymphocyte reaction (MLR) was used to detect the effect of DCsRev on reactive T cells. Results The results confirmed that CTLA4Ig gene was successfully transfected into DCs. The number of DCsRev and the pretreatment of reactive T cells with DCsRev were correlated with the inhibition of T cell proliferation in MLR. When the number of DCs-Rev was between 103 and 104, the inhibitory rate of DCs-Rev was the highest, which was 69.12%. The DCsRev pretreated reactive T cells for 12h had the highest inhibitory rate of T lymphocyte proliferation in MLR 98.3%, 12h later, with the extension of pretreatment time, the inhibition rate has been declining. Splenic lymphocytes induced by DCsRev in vivo proliferate slowly in MLR. Conclusion DCs transfected with CT-LA4Ig not only lose the ability to stimulate MLR, but also inhibit the proliferation of reactive T cells in MLR, suggesting that DCsRev may induce the immune tolerance of antigen-specific T cells.