体外光化学疗法对皮肤移植小鼠产生调节性T细胞的影响

摘要

目的：探讨输注体外光化学法处理的脾淋巴细胞对皮肤移植受体小鼠产生调节性T细胞（Treg）及移植物存活时间的影响。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）。根据受者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。观察受体移植物的存活时间，检测受体外周血中CD4＋CD25＋Foxp3＋Treg表达情况。结果皮肤移植术后，PUVA-BSP组和PUVA-CSP组的受体小鼠外周细胞CD4＋CD25＋Foxp3＋Treg的比例明显高于输注BSP组、CSP组和PBS对照组；PUVA-CSP组高于PUVA-BSP，BSP组和CSP组低于PBS对照组。PUVA-BSP组和PUVA-CSP组的受者小鼠移植皮片存活时间明显长于BSP组、CSP组和PBS对照组（均为P＜0.05）。结论输注足够数量的PUVA-SP可诱导受体体内产生较多的CD4＋CD25＋Foxp3＋Treg，可以显著延长移植物的存活时间。%Objective To investigate the effect of infusion of spleen lymphocytes treated by extracorporeal photochemotherapy on the regulatory T cell (Treg)and survival time of skin allograft in mice. Methods The skin allograft model in mice was established with C57BL/6 mice as donors and BALB/c mice as recipients. The spleen lymphocytes (CSP,BSP)in mice C57BL/6 and BALB/c were isolated,and the mice spleen lymphocytes (PUVA-SP) treated with 8-methoxypsoralen plus ultraviolet (PUVA)were prepared. The experimental animals were randomly divided into 5 groups according to the compositions infused into the recipients through vein:PUVA-BSP,PUVA-CSP,BSP,CSP and phosphate buffer solution (PBS)control groups (n=12 in each group). All recipients of each group were injected with PUVA-BSP,PUVA-CSP,BSP,CSP or PBS on day 7 before the operation,on the operation day and day 7 after the operation through the tail vein,respectively. The survival time of graft in the recipients was observed,and the expression of CD4 +CD25 +Foxp3 +Treg in peripheral blood was detected. Results After skin allograft,the rate of CD4 +CD25 +Foxp3 +Treg in peripheral blood of the recipients in PUVA-BSP group and PUVA-CSP group was significantly higher than those of BSP, CSP and PBS control groups. The rate of CD4 +CD25 +Foxp3 +Treg in PUVA-CSP group was higher than that of PUVA-BSP group,while BSP and CSP groups were lower than that of PBS control group. The survival time of skin graft in the recipients in PUVA-BSP group and PUVA-CSP group was significantly longer than that of BSP,CSP and PBS control groups (all P<0.05 ). Conclusions Sufficient infusion of PUVA-SP can induce more CD4 +CD25 +Foxp3 +Treg in the recipients and prolong survival time of skin graft significantly.