PTDmFoxp3抑制小鼠皮肤移植排斥反应探究.docVIP

PTDmFoxp3抑制小鼠皮肤移植排斥反应探究 　 作者：舒新军， 季宝菊， 蔺昕， 许逊， 田雨香 俆三荣， 邵启祥 【摘要】 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 transduction 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（Plt;0.05）。PTDmFoxp3组，CsA组移植物的存活时间较各对照组显著延长（Plt;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。 【关键词】 小鼠Foxp3融合蛋白； 免疫抑制； 移植排斥 ［Abstract］ Objective: To investigate the potential inhibiting ability to active lymphocytes proliferation and skin allograft rejection of the fusion protein of mouse forkhead box P3 combining with protein transduction domain(PTDmFoxp3) in mice.Methods： The singlecell suspension of lymphocytes was prepared from mouse spleen. Polyclonal activator Con A was added to active the lymphocytes. Then the singlecell suspension of lymphocytes was incubated with different concentrations of PTDmFoxp3，PTDGFP，cyclosporine, and blank control for 48h, respectively. The bioactivity of inhibiting lymphocytes proliferation of each group was detected by incorporating of CCK8 in vitro. Allogeneic fullthickness grafts from Balb/c mice were transplanted onto the back of C57BL/6 mice. Forty C57BL/6 mice were randomly divided into four groups, with each group having 10 mice and receiving a different treatment including PTDmFoxp3, cyclosporine, 0.9% physiological saline injection and PTDGFP, respectively. On the same day of the skin transplantation, the treating regents were injected into C57BL/6 mice abdomen and did continuous injection for another 7 days. Two mice of each group were selected out randomly for histological observation till 9th day after