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目的:研究带有蛋白穿膜结构域的小鼠Foxp3融合蛋白(mouse Foxp3 fusin protein combining with transduc-tion domain,PTD-mFoxp3)对小鼠脾淋巴细胞活化增殖能力和皮肤移植排斥反应的影响。方法:取健康C57BL/6小鼠脾淋巴细胞,在体外经刀豆蛋白A(concanavaline A,ConA)活化,并分别给予环孢素A(cyclosporine,CsA)、不同浓度的PTD-mFoxp3,mFoxp3,PTD-GFP和培养基处理,检测脾淋巴细胞增殖指数。在此基础上进行从Balb/c小鼠到C57BL/6小鼠的皮肤移植试验,将40只C57BL/6受体小鼠随机分为4组,每组10只,分别以PTD-mFoxp3,CsA,0.9%氯化钠注射液(NS),PTD-GFP于手术当天开始连续腹腔注射8天为干预手段。术后第9天各组随机取2只移植小鼠,采集皮片标本,进行组织学检查。其余的8只用于观察移植皮瓣的存活时间。结果:PTD-mFoxp3和CsA相似,与其他处理方法相比明显抑制ConA活化的小鼠脾淋巴细胞增殖指数(P<0.05)。PTD-mFoxp3组,CsA组移植物的存活时间较各对照组显著延长(P<0.05);病理检查显示PTD-mFoxp3组和CsA组移植物淋巴细胞浸润明显轻于各对照组。结论:PTD-mFoxp3能抑制ConA活化的T淋巴细胞的增殖,且具有抑制小鼠皮肤移植排斥反应的作用。
Objective: To investigate the effect of mouse Foxp3 fusin protein combining with transduc-tion domain (PTD-mFoxp3) on the activation and proliferation of mouse splenic lymphocytes and the skin allograft rejection. Methods: Splenic lymphocytes from healthy C57BL / 6 mice were activated by concanavaline A (Con A) in vitro and were given cyclosporine (CsA), PTD-mFoxp3, mFoxp3, PTD-GFP and culture medium to detect spleen lymphocyte proliferation index. On this basis, skin transplantation experiments from Balb / c mice to C57BL / 6 mice were performed. Forty C57BL / 6 mice were randomly divided into 4 groups with 10 mice in each group. PTD-mFoxp3, CsA , 0.9% sodium chloride injection (NS), PTD-GFP on the day of surgery began continuous intraperitoneal injection for 8 days intervention. On the 9th day after operation, 2 mice were randomly selected from each group, and the skin specimens were collected for histological examination. The remaining 8 were used to observe the graft flap survival time. Results: Similar to CsA, PTD-mFoxp3 significantly inhibited splenic lymphocyte proliferation index (P <0.05) of ConA-activated mice compared with other treatments. The survival time of PTD-mFoxp3 group and CsA group was significantly longer than that of the control group (P <0.05). The pathological examination showed that the infiltration of lymphocytes in PTD-mFoxp3 group and CsA group was lighter than that of the control group. Conclusion: PTD-mFoxp3 can inhibit the proliferation of ConA-activated T lymphocytes and inhibit the rejection of skin graft in mice.