敲除BRCC3加重异基因造血干细胞移植受体小鼠急性移植物抗宿主病  

Knockout of BRCC3 aggravates acute GVHD in allogeneic hematopoietic stem cell transplant recipient mice

作　　者：李欣[1] 赵珂 孙慧颖 任广明 高慧英 李长燕 宁红梅 Li Xin;Zhao Ke;Sun Huiying;Ren Guangming;Gao Huiying;Li Changyan;Ning Hongmei(Department of Hematology,Fifth Medical Center of PLA General Hospital,Fifth School of Clinical Medicine,Anhui Medical University,Hefei 230032,China;Beijing Institute of Lifeomics,Academy of Military Sciences,Beijing 100850,China)

机构地区：[1]安徽医科大学解放军总医院第五医学临床学院血液科,合肥230032 [2]中国人民解放军军事科学院军事医学研究院生命组学研究所,北京100850

出　　处：《中华放射医学与防护杂志》2022年第6期401-407,共7页Chinese Journal of Radiological Medicine and Protection

基　　金：蛋白质组学国家重点实验室开放课题(SKLP-O201804)

摘　　要：目的:研究BRCC3的缺失对急性移植物抗宿主病(aGVHD)的影响,并初步探讨其机制。方法:12只C57BL/6J小鼠作为受体,分为野生型(WT组)和BRCC3 -/-型(KO组),每组6只。采用两次4.5 Gy ^(60)Co全身照射(TBI),两次照射间隔30 min,6 h后通过尾静脉输注BABL/c小鼠1×10^(7)骨髓细胞和8×10^(6)脾脏细胞建立aGVHD小鼠模型。在aGVHD小鼠模型中特异性敲除BRCC3,通过组织病理学检测靶器官损伤;酶联免疫吸附试验(ELISA)和细胞因子微球检测技术(CBA)检测小鼠血清中细胞因子的水平;移植后第9天分离脾脏、肝脏和小肠淋巴细胞,运用流式细胞术检测靶器官中T细胞的浸润和活化。 结果:受体小鼠BRCC3的缺失可导致aGVHD小鼠生存期明显缩短( P<0.05);靶器官肝脏损伤明显加重;脾脏中CD8+ T细胞和CD8+CD25+ T细胞的比例明显升高( t=6.53、5.52, P<0.05);肝脏中CD8+ T细胞和CD8+CD25+ T细胞的比例明显升高( t=3.74、3.19, P<0.05)以及小肠中CD8+ T细胞、CD8+CD25+ T细胞和CD8+CD69+ T细胞的比例明显升高( t=3.52、4.06、3.29, P<0.05)。 结论:本研究初步表明BRCC3在受体中的缺失可促进供体CD8+ T细胞的增殖与活化而加重aGVHD,为aGVHD探索了潜在的预防和治疗靶点。Objective To investigate the effect and underlying mechanism of BRCC3 knockout on acute GVHD(aGVHD)of mice.Methods A total of 12 recipient C57BL/6J mice were divided into two groups,including 6 wild type(WT)and BRCC3-/-(KO).The recipients were exposed to 4.5 Gy+4.5 Gy ^(60)Coγ-rays in total body irradiation(TBI)at 30 min intervals.At 6 h post-irradiation,1×10^(7)bone marrow cells and 8×10^(6) splenocytes from BALB/c mice were infused into C57BL/6J mouse via tail vein to develop aGVHD mouse model.BRCC3 was specifically knocked out in aGVHD mouse model.The organ damage was examined through histopathology.The levels of serum cytokines were measured by enzyme-linked immuno sorbent assay(ELISA)and cytometric bead array(CBA),respectively.Spleen,liver and small intestine lymphocytes were isolated at 9 d post-transplantation,and the infiltration and activation of T cells in the target organs were assayed using flow cytometry.Results The absence of BRCC3 in recipient mice significantly shortened survival(P<0.05)with increased liver injury of aGVHD mice.In BRCC3-/-recipient mice,the proportions of CD8+T cells and CD8+CD25+T cells were significantly higher than those in the spleen(t=6.53,5.52,P<0.05),and the proportions of CD8+T cells and CD8+CD25+T cells were significantly increased in the liver(t=3.74,3.19,P<0.05).Similarly,the proportions of CD8+T cells,CD8+CD25+T cells and CD8+CD69+T cells were significantly elevated in the small intestine(t=3.52,4.06,3.29,P<0.05).Conclusions BRCC3 deletion increased the proliferation and activation of donor CD8+T cells and aggravated aGVHD,which might provide a new prevention and treatment target for aGVHD.

关 键 词：BRCC3 急性移植物抗宿主病 T细胞活化 治疗靶点 

分 类 号：R593[医药卫生—内科学]