免疫相关性全血细胞减少患者骨髓补体水平及调节性T细胞数量及功能研究

发布时间：2018-05-10 08:22

本文选题：血细胞减少 + 免疫相关　；参考：《天津医科大学》2009年硕士论文

【摘要】： 目的:了解免疫相关性全血细胞减少(immuno-related pancytopenia,IRP)患者骨髓液补体水平的变化及调节性T细胞(regulatory T cells)数量和功能状况,探讨补体在IRP患者骨髓造血细胞破坏机制中的作用以及调节性T细胞在IRP免疫发病机制中的作用。方法:研究对象为124例IRP患者,其中初治患者(初治组)59例、经激素免疫抑制及促造血治疗后血象恢复正常患者(恢复组)65例及正常对照24名。采用ELISA法测定其骨髓上清中的补体膜攻击复合物(C5b-9)、补体总溶血活性(CH50)、补体C3、C4水平及调节性T细胞相关的细胞因子白细胞介素(interleukin,IL)-2、TGF-β的水平,并使用流式细胞术(FCM)检测IRP患者骨髓单个核细胞膜抗体(CD34~+细胞、Glyco-A~+细胞、CD15~+细胞膜表面自身抗体IgG、IgM)的结合率及调节性T细胞数量(CD4~+CD25~+/CD4~+细胞)、调节性T细胞激活状态(CD4~+CD25~+CD127~-/CD4~+细胞);RT-PCR方法检测其骨髓单个核细胞调节性T细胞转录因子FoxP3和Galectin-10的mRNA表达。结果: 1.IRP初治组患者骨髓C5b-9水平(119.82±53.95)明显高于恢复组(100.74±33.42)和正常对照组(93.86±28.81)(p均＜0.05),而恢复组患者骨髓C5b-9与正常对照组比较差异无统计学意义(p＞0.05);IRP初治组、恢复组患者骨髓CH50水平(33.29±11.51,30.77±10.34)高于正常对照组(24.09±6.37)(p＜0.05),初治组高于恢复组,但差异无统计学意义(p＞0.05);IRP初治组、恢复组患者骨髓补体C3水平(4.90±2.19,4.95±3.47)明显低于正常对照组(6.98±5.56)(p＜0.05),初治组患者骨髓补体C3与恢复组比较差异无统计学意义(p＞0.05);补体C4水平在IRP各组间比较差异无统计学意义(p值均＞0.05);血清补体水平检测结果与骨髓上清中结果一致。骨髓C5b-9与CH50水平呈显著正相关(r=0.241,p=0.003),骨髓CH50与C3水平呈显著负相关(r=-0.303,p=0.007)。IRP患者初治组与恢复组骨髓造血细胞膜抗体IgG型占55.17%,IgM型占54.31%,IgG+IgM型占31.90%,含IgM阳性占86.21%。IRP患者IgM阳性组与IgM阴性组C5b-9水平比较,IgM阳性组高于IgM阴性组(p＜0.05)。骨髓补体C5b-9水平与CD34~+-IgG阳性率、CD34~+-IgM阳性率均呈显著正相关。(r=0.593,p=0.000;r=0.326,p=0.049)。骨髓补体CH50水平与网织红细胞比例呈显著正相关(r=0.421,p=0.000),与间接胆红素呈显著正相关(r=0.230,p=0.032),与血红蛋白、白细胞、血小板、骨髓红系比例均无相关(p均＞0.05)。我们观察了8例IRP患者激素、免疫抑制治疗前后各指标的变化,网织红细胞(reticulocytic Ret)治疗后降至正常(p=0.038),WBC治疗后升高至正常(p=0.002),骨髓补体C3水平治疗后升高(p=0.003),治疗后CD34~+-IgG、CD34~+-IgM阳性率均降低(p=0.016、p=0.022)。 2.IRP初治组、恢复组患者骨髓IL-2水平(5.64±1.70,6.19±2.53)显著低于正常对照组(7.91±3.71)(p＜0.05),初治组低于恢复组,但差异无统计学意义(p＞0.05);IRP初治组、恢复组患者骨髓TGF-β水平(1.79±0.67,1.86±0.77)显著低于正常对照组(2.48±0.94)(p＜0.05),初治组低于恢复组,但差异无统计学意义(p＞0.05);IRP初治组患者骨髓中CD4~+CD25~+/CD4~+细胞数量(22.46±9.47)明显减低,低于正常对照组(30.59±8.58)(p均＜0.05),而恢复组与正常对照组比较差异无统计学意义(p＞0.05);IRP初治组患者骨髓CD4~+CD25~+CD127~-/CD4~+细胞数量(7.18±2.72)明显低于正常对照组(10.44±3.24)(p＜0.05),而恢复组与正常对照组比较差异无统计学意义(p＞0.05);初治组、恢复组、正常对照组三组FoxP3 mRNA的相对表达量分别为(0.34±0.25)、(0.69±0.51)和(0.82±0.65),初治组明显低于恢复组和正常对照组(p均＜0.05),初治组、恢复组、正常对照组Galectin-10 mRNA的相对表达量分别为(0.66±0.11)、(0.74±0.11)和(0.76±0.09),初治组明显低于恢复组和正常对照组(p均＜0.05)。结论:IRP患者特别是自身抗体为IgM者其外周血细胞减少与骨髓单个核细胞自身抗体激活补体破坏造血细胞有关;调节性T细胞的数量和功能异常在IRP免疫发病机制中发挥重要作用。
[Abstract]:Objective: To investigate the changes in the complement level of bone marrow fluid in patients with Immuno-related pancytopenia (IRP) and the number and function of regulatory T cells (regulatory T cells), and to explore the use of complement in the destruction mechanism of bone marrow hematopoietic cells in IRP patients and the role of regulatory T cells in the pathogenesis of IRP. Use.
Methods: the subjects were 124 patients with IRP, of which 59 cases were first treated (initial treatment group), 65 cases and 24 normal controls were restored to normal patients after hormone immunosuppression and hematopoiesis. The complement membrane attack complex (C5b-9), total complement hemolytic activity (CH50), complement C3, C4 level were measured by ELISA method. Regulatory T cell related cytokines interleukin (IL) -2, TGF- beta level, and the use of flow cytometry (FCM) to detect the membrane antibody of bone marrow mononuclear cells in IRP patients (CD34~+ cells, Glyco-A~+ cells, CD15~+ cell membrane surface autoantibody IgG, IgM) and regulating cell number The activated state of the ganglion T cells (CD4~+CD25~+CD127~-/CD4~+ cells), and the RT-PCR method to detect the mRNA expression of the regulatory T cell transcription factor FoxP3 and Galectin-10 in the mononuclear cells of the bone marrow.
Result:
The bone marrow C5b-9 level of the 1.IRP group (119.82 + 53.95) was significantly higher than that in the recovery group (100.74 + 33.42) and the normal control group (93.86 + 28.81) (P < 0.05), while the bone marrow C5b-9 in the recovery group was not significantly different from that of the normal control group (P > 0.05), and the level of the bone marrow CH50 in the recovery group was higher than that of the recovery group (33.29 + 11.51,30.77 + 10.34). In the normal control group (24.09 + 6.37) (P < 0.05), the initial treatment group was higher than the recovery group, but the difference was not statistically significant (P > 0.05). The level of bone marrow complement C3 (4.90 + 2.19,4.95 3.47) in the recovery group was significantly lower than that of the normal control group (6.98 + 5.56) (P < 0.05). There was no significant difference between the bone marrow complement C3 and the recovery group in the primary treatment group (P > P >). 0.05): there was no significant difference in the level of complement C4 between the groups of IRP (P > 0.05); the results of serum complement level were consistent with the results of bone marrow supernatant. There was a significant positive correlation between bone marrow C5b-9 and CH50 level (r=0.241, p=0.003), and a significant negative correlation between the bone marrow CH50 and C3 level (r=-0.303, p=0.007) and the bone marrow of the primary and recovery groups of the.IRP patients. The hematopoietic cell membrane antibody IgG type accounted for 55.17%, the IgM type accounted for 54.31%, the IgG+IgM type accounted for 31.90%, the IgM positive group was compared with the C5b-9 level in the IgM positive group and the IgM negative group, and the IgM positive group was higher than the IgM negative group (P < 0.05). 26, p=0.049). The level of bone marrow complement CH50 was positively correlated with the ratio of reticulocyte (r=0.421, p=0.000). There was a significant positive correlation with indirect bilirubin (r=0.230, p=0.032). There was no correlation with hemoglobin, leukocyte, platelets, and bone marrow erythroid ratio (P > 0.05). We observed 8 cases of IRP patients hormone, and all indexes before and after immunosuppressive therapy. After treatment, the reticulocyte (reticulocytic Ret) was reduced to normal (p=0.038) after treatment. After WBC treatment, it increased to normal (p=0.002), and after C3 level of bone marrow complement increased (p=0.003). The positive rate of CD34~+-IgG and CD34~+-IgM decreased (p=0.016, p=0.022) after treatment.
2.IRP in the primary treatment group (5.64 + 1.70,6.19 + 2.53) in the recovery group was significantly lower than that in the normal control group (7.91 + 3.71) (P < 0.05), and the initial treatment group was lower than the recovery group, but the difference was not statistically significant (P > 0.05). The level of TGF- beta in the recovery group was significantly lower than that of the normal control group (1.79 + 0.67,1.86 + 0.77) (2.48 + 0.94) (P < 0.05) (P < 0.05). The initial treatment group was lower than the recovery group, but the difference was not statistically significant (P > 0.05). The number of CD4~+CD25~+/CD4~+ cells in the bone marrow (22.46 + 9.47) in the initial IRP group was significantly lower than that in the normal control group (30.59 + 8.58) (P < 0.05), but there was no significant difference between the recovery group and the normal control group (P > 0.05), and the bone marrow CD4~+CD25 in the primary treatment group of IRP was CD4~+CD25. The number of ~+CD127~-/CD4~+ cells (7.18 + 2.72) was significantly lower than that in the normal control group (10.44 + 3.24) (P < 0.05), but there was no significant difference between the recovery group and the normal control group (P > 0.05). The relative expression of FoxP3 mRNA in the early treatment group, the recovery group and the normal control group was (0.34 + 0.25), (0.69 + 0.51) and (0.82 + 0.65), and the primary treatment group was significantly lower. In the recovery group and the normal control group (P < 0.05), the relative expression of Galectin-10 mRNA in the primary treatment group, the recovery group and the normal control group was (0.66 + 0.11), (0.74 + 0.11) and (0.76 + 0.09), and the primary treatment group was significantly lower than the recovery group and the normal control group (P < 0.05).
Conclusion: the decrease of peripheral blood cells in patients with IRP, especially autoantibodies of IgM, is related to the destruction of hematopoietic cells by autoantibody activating complement of bone marrow mononuclear cells, and the number and dysfunction of regulatory T cells play an important role in the mechanism of IRP immunization.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392

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