同种异体肢体移植特异性免疫耐受诱导方案(英文)

Induction program of donor-specific immune tolerance in limb allotransplantation

背景:通过诱导受体产生免疫耐受能够解决移植领域存在的许多问题,如长期应用免疫抑制剂带来的毒副作用,慢性排斥反应等。目的:观察联合应用亚致死量放射线照射、氟达拉滨及骨髓腔内骨髓移植对诱导大鼠同种异体肢体移植产生特异性免疫耐受的影响。设计、时间及地点:随机同期对照动物实验,于2004-01/2008-12在哈尔滨医科大学完成。材料:40只雄性Wistar大鼠随机分成4组:单纯移植组、氟达拉滨组、骨髓移植组、氟达拉滨+骨髓移植组,每组各10只。40只SD大鼠为骨髓移植供体。供体DA大鼠用于皮肤移植实验。方法:制作同种异体右后肢移植模型,单纯移植组仅进行肢体移植;氟达拉滨组在肢体移植前1d,受体给予亚致死量60Co照射及腹腔内注射氟达拉滨50mg/kg;骨髓移植组肢体移植当天受体接受供体骨髓腔内骨髓移植1×1012L-1,10μL;氟达拉滨+骨髓移植组受体联合应用亚致死量照射、注射氟达拉滨及骨髓腔内骨髓移植。主要观察指标:观察移植物排斥反应征象及存活情况,并对氟达拉滨+骨髓移植组产生免疫耐受大鼠进行SD大鼠及DA大鼠皮肤移植及脾细胞中细胞因子mRNA检测。结果:与其他实验组比较,氟达拉滨+骨髓移植组移植物发生排斥反应的时间及存活时间均显著延长(P<0.01)。氟达拉滨+骨髓移植组大鼠仅对第3方DA大鼠的皮肤呈现强烈的免疫反应,移植皮肤初为水疱、渗出,逐渐溃疡、糜烂、焦痂、变黑坏死。与单纯移植组比较,氟达拉滨+骨髓移植组大鼠Th1型细胞因子的表达明显降低,而Th2型细胞因子的表达明显增高。结论:氟达拉滨与骨髓移植联合方案可以诱导大鼠同种异体肢体移植产生特异性免疫耐受。

BACKGROUND：Immune tolerance by inducing recipients can solve many problems in the field of transplantation,including toxic and side effects due to long-term application of immunosuppressive agent,as well as chronic rejection.OBJECTIVE：To observe effects of combined application of sublethal dose radiographic exposure,fludarabine and bone marrow transplantation in the marrow cavity on specific immunologic tolerance in rats undergoing limb allotransplanta-tion.DESIGN,TIME AND SETTING：The randomized concurrent controlled animal study was performed at the Harbin Medical University from January 2004 to December 2008.MATERIALS：A total of 40 male Wistar rats were equally and randomly assigned to 4 groups,transplantation alone group,fludarabine group,bone marrow transplantation group,fludarabine ＋ bone marrow transplantation group.A total of 40 Sprague Dawley rats served as donors.Dark Agouti rats（donors）were used in skin transplantation study.METHODS：Models of allogenic right hindlimb transplantation were created.Rats in the transplantation alone group received limb transplantation.Rats in the fludarabine group underwent 60Co irradiation and intraperitoneal injection of 50 mg/kg fludarabine at day 1 prior to limb transplantation.Rats in the bone marrow transplantation group were subjected to bone marrow transplantation（1×1012/L,10 μL）on the day of limb transplantation.Rats in the fludarabine ＋ bone marrow trans-plantation group received combined application of sublethal dose radiographic exposure,fludarabine and bone marrow transplantation in the marrow cavity.MAIN OUTCOME MEASURES：The condition of survival and rejection of grafts was observed.In the fludarabine ＋ bone marrow transplantation group,tolerance rats received Sprague Dawley rats and Dark Agouti rats skin grafting and cytokine mRNA was detected in splenocytes.RESULTS：Compared with other experimental groups,survival time and rejection duration were significantly prolonged in the fludarabine ＋ bone marrow transplantation group（P〈0.01）.Rats in the fludarabine ＋ bone marrow transplantation group only presented strong immune reaction to third party Dark Agouti rats.Transplanted skin presented blister,exudation,gradually ulcer,erosion,eschar,darkening and necrosis.Compared with transplantation alone group,Th1 type cytokine expression was significantly decreased in the fludarabine ＋ bone marrow transplantation group,but Th2 type cytokine ex-pression was significantly increased.CONCLUSION：Combination of fludarabine and bone marrow transplantation can induce specific immune tolerance in limb allotransplantation in rats.