体外光化学疗法处理的调节性树突细胞对T细胞增殖的影响

目的 探讨体外光化学疗法处理的调节性树突细胞对T细胞增殖的影响.方法 获取人外周血单个核细胞(PBMC),利用白介素-4重组人粒细、巨噬细胞集落刺激因子诱导生成未成熟的树突细胞(imDC).分离获取人脾脏淋巴细胞(SP),经光化学疗法(PUVA)处理后获得PUVA-SP;将imDC与PUVA-SP共培养得到体外光化学治疗性树突细胞(ecpDC);将imDC与SP共培养得到SPDC;imDC加入10ng/ml的脂多糖(LPS),培养1d后得到DC.收集4组细胞,检测其表面抗体CD11c、CD83、CD86的表达.采用混合淋巴细胞培养的方法 检测吞噬了供者凋亡的脾淋巴细胞后的受者imDC对受者T细胞增殖的影响.结果 经PUVA处理后细胞早期凋亡率为91.33%.表面分子CD83、CD86在ecpDC的阳性表达率为分别为22.83%±5.26%、22.06%±4.37%,与在imDC的阳性表达率(分别为15.06%±0.59%、15.19%±1.83%)相近(P>0.05),但明显低于在DC的阳性表达率(分别为99.79%±0.36%、99.85%±0.19%),差异有统计学意义(P<0.01).吞噬了供者凋亡的脾淋巴细胞后的受者imDC细胞可明显抑制受者T细胞的增殖.结论 体外光化学疗法诱导的凋亡脾淋巴细胞可抑制树突细胞的成熟,并可抑制受者T淋巴细胞的增殖.

Influence of extracorporeal photochemotherapy-treated regulatory dendritic cells on T cell proliferation

Objective To investigate the effect of regulatory dendritic cells treated by extracorporeal photochemotherapy on T cell proliferation. Methods Human peripheral blood mononuclear cells (PBMCs) were obtained and the immature dendritic cells (imDCs) were induced by recombinant human granulocyte and macrophage colony stimulating factors. SPDCs were obtained by PUVA treatment, and ECDCs were co-cultured with imDCs and PUVA-SP to obtain immunoprecipitated dendritic cells. In vitro, imDCs were co-cultured with SPDCs to obtain SPDCs; imDCs were added to 10ng/ml of LPS, and cultured for 1 day to obtain DCs. The expressions of CD11c, CD83 and CD86 on the surface of the cells were detected. The effect of imDCs on the proliferation of recipient T cells was detected by mixed lymphocyte culture method. Results The early apoptosis rate of PUVA-treated cells was 91.33%. The positive expression rates of CD83 and CD86 in ecpDCs were 22.83%±5.26% and 22.06%±4.37%, respectively, which were similar to those of imDCs (15.06%±0.59%, 15.19%±1.83% (P<0.01), but significantly lower than those in DCs (99.79%±0.36%, 99.85%±0.19%, respectively), the difference was statistically significant (P<0.01). The recipient imDC cells phagocyting the appoptotic splenic lymphocytes from the donor significantly inhibited the proliferation of recipient T cells. Conclusion Apoptosis of splenic lymphocytes induced by extracorporeal photochemotherapy can inhibit the maturation of dendritic cells and inhibit the proliferation of T lymphocytes.