探讨抗γ公共链单抗诱导移植免疫耐受的作用机制  

Mechanism of induction of transplant tolerance via γc signals blockade in murin e model

作　　者：昌盛[1] 沈世乾[1] 陈必成[1] 张鑫[1] 杜敦峰[1] 周洁[1] 陈忠华 

机构地区：[1]华中科技大学同济医学院附属同济医院器官移植研究院,教育部,卫生部器官移植重点实验室,武汉430030

出　　处：《中华器官移植杂志》2005年第8期470-473,共4页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金资助项目(30271243)

摘　　要：目的探讨抗γ公共链单抗在诱导移植免疫耐受中的作用机制。方法将实验动物分为2组,每组20只。实验组:在Balb/c裸小鼠(H-2d)体内用CFSE标记的Balb/c小鼠(H-2d)T细胞(3×107个)重建其免疫系统,同时给予经丝裂霉素C处理的供者C57BL/6小鼠(H-2b)脾细胞(5×106个)静脉输注,24h后实验组受鼠经腹腔注射抗γc单克隆抗体(4G3,3E12,TUGm2,各0.5mg)和抗白细胞介素2受体(IL-2R)β链单克隆抗体(TM-β1,0.5mg)混合液;对照组:以IgG2a2.0mg作为同型对照代替抗体腹腔注射,余处理与实验组相同。经上述处理后12h和48h后处死受鼠,取其脾细胞及外周血,利用流式细胞技术检测T细胞的增殖及凋亡。结果经抗体处理12h和48h后,实验组T细胞未发生明显增殖;而对照组两时间点均可观察到CFSE标记细胞荧光强度明显地减低,表明T细胞发生了分裂增殖。同时AnnexinV法可见,实验组标记有CD3-PE单抗的T细胞12h发生了明显的凋亡,但48h却几乎无明显的凋亡细胞;而对照组两时间点则均未检测到明显的细胞凋亡。两组间12h结果差异有统计学意义(P<0.01),但48h差异无统计学意义(P>0.05)。结论移植术前给予抗γ公共链单抗联合供者细胞特异性输注,通过有效抑制针对供者抗原发生效应的T细胞增殖,可进一步促使其发生凋亡而被有效清除,从而最终实现免疫耐受,且此种耐受具有抗原特异性。Objective To investigate the possible mechanism of induction of transplant tolerance via common γ chain （γc）. Methods Splenocytes （5 × 10^6 cells） from C57BL/6 （H-2^b） mice were har vested and transfused into T-cell deficient Balb/c （H-2^d） nude mice that were reconstituted with syngeneic T-cells （3 × 10^7 cells from wild-type.Balb/c mice） labeled with CFSE. On the day 2, recipients received i. p. injection of mixture of anti-IL-2Rβ mAb （TM-β1, 0. 5 rag） and anti-γc mAbs （i. e. , 4G3, 3E12 and TUGm2, 0. 5 rag, respectively）, or no mAbs treatment instead of IgG2a, 2.0 mg （isotype control group）. The labeled T-cells were isolated and harvested from recipient spleen after 12 h or 48 h. T-cell proliferation was examined with fluorescent dye labeling technique and T-cell apoptosis was detected by Annexin V staining. Flow cytometry was used to analyze the results. Results T-cell proliferation was markedly inhibited and apoptotic T cells could be detected 12 h after the mAbs injection. The proliferation was constantly inhibited, but no more apoptotic T-cells were found in 48 h. In control group, however, T-cells actively proliferated and no apoptosis was detected at both time points. Conclusions This result may be associated with inhibition of antigen-specific T-cell proliferation and induction of apoptosis. We anticipate that this protocol may develop a novel approach to induce donor-specific tolerance.

关 键 词：抗γ公共链单抗 移植 免疫耐受 作用机制 丝裂霉素C 

分 类 号：R699.2[医药卫生—泌尿科学]