γδΤ细胞在获得性纯红细胞再生障碍性贫血发病机制中的作用  

Role of γδT Cells in Pathogenesis of Acquired Pure Red Cell Aplastic Anemia

作　　者：刘敏[1] 刘霆[1] 孟文彤[1] 朱焕玲[1] 崔旭[1] 

机构地区：[1]四川大学华西医院血液科血液病研究室,成都610041

出　　处：《中国实验血液学杂志》2007年第1期142-146,共5页Journal of Experimental Hematology

基　　金：四川省科技厅课题资助(编号:01SY051-39)

摘　　要：本研究检测获得性纯红细胞再生障碍性贫血(acquired pure red cell aplastic anemia,A-PRCA)患者γδT细胞亚群数量和功能变化,探讨γδΤ细胞在A-PRCA发病机制中的作用。对11例经骨髓涂片和活检确诊的A-PRCA患者,给予环胞菌素A和雷公藤多甙治疗;采用流式细胞术检测免疫抑制剂治疗前后患者T细胞亚群和γδT细胞水平;分离患者外周血中单个核细胞并置于含有10%FCS,PHA10μg/ml,rIL-250U/ml的RPMI1640培养液培养2周,然后用TCRγδ磁珠分选出γδT细胞,在体外与正常人骨髓共同培养,观察对红系、粒系集落生长的影响。结果显示,治疗前组外周血CD3+/CD8+细胞数较正常对照组升高,CD4+/CD8+比例倒置(P<0.05);治疗前组患者γδT细胞水平有明显升高(P<0.05)。治疗后,有效组CD3+/CD8+细胞数下降(P<0.05),CD4+/CD8+比值上升(P<0.01);γδT细胞水平较治疗前有明显降低(P<0.05)。从患者外周血分离的γδT细胞体外能抑制正常骨髓红系CFU-E和BFU-E生长,并随浓度的增长抑制作用越明显,而在不同的浓度梯度下对粒系生长无明显影响。结论:A-PRCA患者γδT细胞亚群增高,其在PRCA的发病机制中起了一定作用;体外分离培养的患者γδT细胞可抑制正常红系集落生长,对粒系集落生长影响不明显;环胞素A治疗A-PRCA有较好疗效。This study was purposed to investigate the changes in quantum and function of γδT cell subsets, and to explore its significance in pathogenesis of acquired pure red cell aplastic anemia（A-PRCA）. Eleven patients were diagnosed as A-PRCA based on bone marrow smear and biopsy, and were treated with cyclosporine A and glucosidorum tripterygll totorum. The flow cytometry technique was used for analyses of T cells subsets and γδT cells. Furthermore, peripheral mononuclear cells（MNC） isolated from A-PRCA patients were cultured in RPMI 1640 medium （ 105 cellsdml） containing 10% FCS, phytohemagglutinin（ PHA, 10 μg/ml）, and recombinant human interleuldn-2（ rIL-2, 50 U/ml） for two weeks, then γδT cells were isolated with the TCRγδ Microbead Kit from cultured cells. The collected γδT cells were incubated with normal control bone marrow MNC in RPMI 1640 medium（ 37℃, 5% CO2 atmosphere） for CFU-E, CFU-GM, and BFU-E colony assay. The result showed that compared with the control group, CD3 ^＋ , CD8 ^＋ cells increased significantly in the patient group（ P 〈 0.05 ）, the CD4 ^＋/CD8 ^＋ ratio decreased and reversed, and γδT cells were significantly increased in patient group（ P 〈 0.05 ）. After treatment with cyclosporine A, 9 out of 11 patients got good response, and CD3 ^＋ , CD8 ^＋ cells in the responding patient decreased, the ratio of CD4 ^＋/CD8 ^＋ returned to normal, and γδT cells also decreased to normal range. Moreover, in vitro culture, the γδT cells isolated from A-PRCA patients showed an inhibiting acdon to CFU-E and BFU-E but not to CFU-GM in a dose-dependent manner. It is concluded that γδT cells increase in A-PRCA patients, and decrease in parallel to normal range with significant improvement of anemia symptoms after immune suppressive therapy. The γδT cells isolated from A-PRCA patients showed an inhibiting action to CFU-E and BFU-E but not to CFU-GM in vitro culture, suggesting that γδT cells may bring an impact on the research of A-PRCA pathoge

关 键 词：获得性纯红细胞再生障碍性贫血 细胞免疫 ΓΔT细胞 

分 类 号：R556.5[医药卫生—血液循环系统疾病]