论文部分内容阅读
目的探讨Th1/Th2细胞平衡偏离及平衡回复对重型再生障碍性贫血(sAA)骨髓CD34+细胞体外扩增和集落生成的影响。方法以1例确诊的sAA患者为研究对象。(1)分离骨髓单个核细胞,用免疫磁珠法富集CD34+细胞、CD4+(Th)细胞。(2)以流式细胞术(FCM)检测CD4+细胞中Th1、Th2细胞比例。(3)扩增CD34+细胞并再次富集以获足量CD34+细胞,分为4组对照组;Th细胞作用组;Th细胞+IFN-γ作用组;Th细胞+IL-4作用组。(4)各组扩增培养10d,继以集落生成试验,测定各组CD34+细胞扩增率和集落产率。(5)患者经免疫抑制治疗后随访,用FCM监测Th1/Th2细胞比。结果(1)患者经5个月治疗获缓解。(2)缓解前、后Th1/Th2比分别是22.47和12.27,正常对照组为8.98±4.45。(3)CD34+细胞扩增率,以对照组最高,其次为Th细胞+IL-4组、Th细胞组,Th细胞+IFN-γ组的最低。(4)各组CD34+细胞集落产率与其扩增率数值平行相关,即对照组最多,其次为Th细胞+IL-4组、Th细胞组,Th细胞+IFN-γ组的最少。结论Th1细胞反应亢进直接抑制sAA患者CD34+细胞在体外的自我更新和增殖分化,IL-4能拮抗这种造血抑制效应,这可能是通过调节Th1/Th2平衡而间接实现的。
Objective To investigate the effects of Th1 / Th2 balance and homeostasis on in vitro expansion and colony formation of bone marrow CD34 + cells in patients with severe aplastic anemia (sAA). Methods One patient with confirmed sAA was selected as the research object. (1) Bone marrow mononuclear cells were isolated and CD34 + cells and CD4 + (Th) cells were enriched by immunomagnetic beads method. (2) The proportion of Th1 and Th2 cells in CD4 + cells was detected by flow cytometry (FCM). (3) CD34 + cells were expanded and re-enriched to obtain sufficient amount of CD34 + cells. The cells were divided into four groups: control group, Th cells, Th cells + IFN-γ group and Th cells + IL-4 group. (4) Each group was expanded and cultured for 10 days, followed by colony formation test to determine the CD34 + cell expansion rate and colony yield in each group. (5) Patients were followed up after immunosuppressive therapy, and the ratio of Th1 / Th2 cells was monitored by FCM. Results (1) Patients were relieved after 5 months of treatment. (2) The Th1 / Th2 ratios before and after remission were 22.47 and 12.27, respectively, and those in the normal control group were 8.98 ± 4.45. (3) The rate of CD34 + cell expansion was highest in the control group, followed by the lowest in the Th + IL-4, Th and IFN-γ groups. (4) The colony yield of CD34 + cells in each group was in parallel with the value of its amplification rate, which was the highest in the control group, followed by the least in the group of Th cells + IL-4, Th cells and Th cells + IFN-γ. Conclusion Th1 hyperactivity directly inhibits the self-renewal and proliferation of CD34 + cells in sAA patients. IL-4 antagonizes this hematopoietic inhibitory effect, which may be indirectly mediated through the regulation of Th1 / Th2 balance.