摘要
目的探讨CD8^+CD45RC^low的调节性T细胞及相关细胞因子在大鼠同种异体原位肝移植耐受过程中的分布规律。方法近交系BN大鼠为供体,雄性Lewis大鼠为受体,双套袖法同种异体原位肝脏移植。实验分为:肝移植短期组10只,肝移植耐受组10只和对照组6只,各组术后常规检测肝功能。苏木精-伊红(HE)染色用于判定移植物达到自然免疫耐受。流式细胞术分析外周血、移植物、淋巴结中淋巴细胞亚群的变化,Luminex技术进行细胞因子定量检测。结果有70%的移植术后大鼠自然免疫耐受。长期耐受组与对照组的谷草转氨酶(AST)、谷丙转氨酶(ALT)比较,差异无统计学意义[(456±54)U/ml比(460±151)U/ml,(122±35)U/ml比(121±46)U/ml,P>0.05]。与早期移植实验组比较,在免疫耐受大鼠外周血中CD8+CD45RClowT细胞显著增加,移肝脏局部随着耐受时间的延长其相对含量不断增加;移植物周围淋巴结中该类细胞的表达差异无统计学意义(P>0.05);外周血中pDC在与对照组比较短期组明显增加。移植物局部细胞因子检测发现干扰素(IFN)-γ在早期升高,随后恢复到正常水平;白细胞介素(IL)-10出现持续升高的表现。结论在大鼠同种异体原位肝移植免疫耐受过程中,CD8^+CD45RC^low调节性T细胞显著升高,并在外周血和肝脏中高表达,通过提高抑制性的细胞因子IL-10对长期免疫耐受起重要调节作用。
Objective To explore the mechanism of CD8^+CD45RC^low regulatory T cells and its related cytokines function during the tolerance process of orthotopic liver transplantation in rats.Methods The male Brown Norway rats were selected as the donor,the Lewis rats were the recipients,the allogenic orthotopic liver transplantation model was established by the double cuff method.Experiment animals were divided into early transplantation group(10 rats),tolerance group(10 rats),sham group.Liver function were detected.The tolerance recipients were detected by hematoxylin and eosin(HE)staining.The Luminex used to detect of cytokine and the flow cytometry were used to analyze the changes of subsets of lymphocytes in the peripheral blood/graft and draining lymph node.Results 70%of recipients in operation group could be induced natural immune tolerance.The levels of aspartate aminotransferase(AST)in tolerance group were[(456±54)U/ml]vs.those of sham group[(460±151)U/ml],(P>0.05);the levels of alanine aminotransferase(ALT)in tolerance group were[(122±35)U/ml]vs.those of sham group[(121±46)U/ml,P>0.05].The expression of CD8^+CD45RC^low T cells in tolerance group were increased than in short term group;in grafts it was up regulated in tolerance group than in sham group,and it was no significance between sham group and tolerance group in lymph node around graft.The expression of pDC in short term group are significantly increased compared with sham group.The interferon(IFN)-γwas increased in short term group,return to normal level in tolerance group.interleukin(IL)-10 in grafts milieu were dramatically increased,keep in high level in tolerance group.Conclusion In the tolerance recipients of rat orthotopic liver transplantation model,CD8^+CD45RC^low regulatory T cells were significantly increased and aggregated in the peripheral blood and liver.The inhibitory cytokines IL-10 was important to maintain the long-term graft tolerance.
作者
白纯
李先亮
杨龙
刘焕业
寇建涛
朱继巧
马军
吕少诚
贺强
Bai Chun;Li Xianliang;Yang Long;Liu Huanye;Kou Jiantao;Zhu Jiqiao;Ma Jun;Lyu Shaocheng;He Qiang(Capital Medical University,Beijing 100069,China;Department of Hepatobiliary,Pancreatic and Splenic Surgery,Beijing Chaoyang Hospital Affiliated to Capital Medical University,Beijing 100020,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第5期849-851,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金面上项目(81571554、81471590、81273270、81041110).
