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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。
In this study, the quantity and function of γδT cell subsets in patients with acquired pure red cell aplastic anemia (A-PRCA) were detected to investigate the role of γδT cells in the pathogenesis of A-PRCA. Cyclosporin A and tripterygium glycosides were given to 11 A-PRCA patients who were diagnosed by bone marrow smear and biopsy. The levels of T lymphocyte subsets and γδT cells before and after immunosuppressive therapy were detected by flow cytometry. Peripheral blood mononuclear cells were isolated and cultured in RPMI1640 medium containing 10% FCS, PHA10μg / ml and rIL-250U / ml for 2 weeks. Then, γδT cells were sorted by TCRγδ magnetic beads and compared with normal human bone marrow Co-cultivation, observation of erythroid and granulocyte colony growth. The results showed that the number of CD3 + / CD8 + cells in the peripheral blood of the pre-treatment group was higher than that of the normal control group, and the ratio of CD4 + / CD8 + was upside down (P <0.05). The levels of γδT cells in the pre-treatment group were significantly increased (P <0.05). After treatment, the number of CD3 + / CD8 + cells in the effective group decreased (P <0.05) and the ratio of CD4 + / CD8 + increased (P <0.01). The level of γδT cells decreased significantly (P <0.05). The γδT cells isolated from the peripheral blood of patients can inhibit the growth of normal bone marrow erythrocytes CFU-E and BFU-E in vitro, and the inhibitory effect of CFU-E and BFU-E is more obvious with different concentration gradients. CONCLUSIONS: The increase of γδT cell subsets in A-PRCA patients plays a role in the pathogenesis of PRCA. The γδT cells isolated in vitro from patients with A-PRCA can inhibit the growth of normal erythroid colonies and have no obvious effect on the growth of granulocyte colony; A-treatment of A-PRCA has a good effect.