体外光分离置换疗法对皮肤移植小鼠IL-12p70和Th1/Th2类细胞因子产生的影响

摘要

目的 探讨输注体外光分离置换疗法处理的脾淋巴细胞对皮肤移植小鼠白细胞介素(IL)-12p70和辅助性T细胞(Th)1/Th2类细胞因子产生的影响.方法 以C57BL/6小鼠为供体,BALB/c小鼠为受体,建立小鼠皮肤移植模型.分离C57BL/6和BALB/c小鼠脾淋巴细胞(CSP、BSP),制备经8-甲氧基补骨脂素联合长波紫外线(PUVA)处理的小鼠脾淋巴细胞(PUVA-SP).根据受体静脉输注的成分将实验动物随机分为5组(每组12只):PUVA-BSP组、PUVA-CSP组、BSP组、CSP组及磷酸盐缓冲液(PBS)对照组,每组受体分别于术前7 d、手术当日和术后7 d按组别从尾静脉注入PUVA-BSP、PUVA-CSP、BSP、CSP或PBS.观察经PUVA处理脾淋巴细胞凋亡情况,检测受体小鼠外周血中IL-12p70和Th1/Th2类细胞因子产生的情况.结果 移植术后,PUVA-BSP组和PUVA-CSP组小鼠外周血的IL-12p70表达水平明显低于BSP组、CSP组和PBS对照组(均为P<0.01);PUVA-BSP组和PUVA-CSP组的Th1类细胞因子IL-2、干扰素(IFN)-γ 表达水平均明显低于BSP组、CSP组和PBS对照组(均为P<0.01);PUVA-BSP组和PUVA-CSP组的Th2类细胞因子IL-10表达水平明显高于BSP组、CSP组和PBS对照组(均为P<0.01).结论 输注足够数量的PUVA-SP可诱导受体体内低表达IL-12p70,并且诱导产生Th2免疫偏移.%Objective To evaluate the effect of extracorporeal photopheresis upon the expression levels of interleukin (IL)-12p70 and T helper cell (Th) 1/Th2-like cytokines in splenic lymphocytes of mouse models undergoing skin transplantation. Methods The C57BL/6 mice were used as the donors and BALB/c mice as the recipients to establish mouse models with skin allograft. The splenic lymphocytes in the C57BL/6 and BALB/c mice (CSP and BSP) were isolated and treated with 8-methoxypsoralen combined long-wave ultraviolet (PUVA-SP). According to the components of intravenous infusion into the recipients, all experimental animals were randomly divided into the PUVA-BSP, PUVA-CSP, BSP, CSP and phosphate buffer solution (PBS) control groups (n=12 for each group). The mice were injected with PUVA-BSP, PUVA-CSP, BSP, CSP or PBS via the caudal vein at preoperative 7 d, upon the day of surgery and at postoperative 7 d, respectively. The apoptosis of the splenic lymphocytes was observed after PUVA treatment. The expression levels of IL-12p70 and Th1/Th2-like cytokines in the peripheral blood of the recipients were quantitatively measured. Results After the skin transplantation, the expression levels of IL-12p70 in the peripheral blood of mice in the PUVA-BSP and PUVA-CSP groups were significantly down-regulated compared with those in the BSP, CSP and PBS control groups (all P<0.01). In the PUVA-BSP and PUVA-CSP groups, the expression levels of Th1-like cytokine IL-2, interferon (IFN)-γwere dramatically lower than those in the BSP, CSP and PBS control groups (all P<0.01). The expression levels of Th2-like cytokine IL-10 in the PUVA-BSP and PUVA-CSP groups were significantly up-regulated compared with those in the BSP, CSP and PBS control groups (all P<0.01). Conclusions Infusion of PUVA-SP at a sufficient dose can induce the low expression level of IL-12p70 and drive the incidence of Th2 immune deviation in the recipient BALB/c mice.