本文選題：免疫相關(guān) + 血細(xì)胞減少��； 參考：《天津醫(yī)科大學(xué)》2015年博士論文

【摘要】：目的檢測免疫相關(guān)性血細(xì)胞減少(immuno-related Immuno-related hemtocytopenia,IRH)患者骨髓造血細(xì)胞膜上抗體免疫球蛋白G(immunoglobulin G,IgG)亞型,檢測骨髓上清IgG各亞型含量,探索IRH患者IgG的優(yōu)勢亞型,為進(jìn)一步揭示IRH患者的發(fā)病機(jī)制提供理論上的依據(jù)。方法1.研究對象為初治IRH患者(實驗組),以意義未名的血細(xì)胞減少癥患者(ICUS)及正常人分別為病例對照組及正常對照組,進(jìn)行以下研究分析:(1)采用流式細(xì)胞術(shù)(FCM)檢測IRH患者骨髓細(xì)胞(CDl5+、CD34+及GlycoA+細(xì)胞)膜抗體(IgM、IgG);(2)FCM檢測IgG(+)的IRH患者及正常人(CDl5+及GlycoA+細(xì)胞)膜抗體(IgG1、IgG3);(3)采用免疫組化方法(IHC)對Ig G(+)的IRH患者,ICUS組及正常對照組骨髓有核細(xì)胞進(jìn)行IgG亞型(IgG1、IgG2、IgG3、IgG4)染色。2.選取骨髓造血細(xì)胞膜抗體IgG陽性初治IRH患者為實驗組,以IRH緩解病人、ICUS初治病人及正常人分別為病例及正常對照組,進(jìn)行下述研究:(1)采用流式微球捕獲技術(shù)(CBA)對骨髓上清IgG亞型(IgG1,IgG2,IgG3,IgG4)含量進(jìn)行檢測;(2)采用FCM對外周血CD5+CD19+/CD19+比例進(jìn)行檢測;(3)RT-PCR的方法檢測外周血淋巴細(xì)胞IgG1的mRNA表達(dá)。結(jié)果1.IRH患者骨髓造血細(xì)胞膜抗體IgG陽性率58.69%(27/46)明顯高于病例對照組0%及正常對照組0%,骨髓造血細(xì)胞膜抗體IgM陽性率80.43%(37/46)。2.實驗組CD15+細(xì)胞IgG1的陽性率明顯高于正常人((3.13±1.80)%vs(0.87±0.38)%),P〈0.05);實驗組CD15+細(xì)胞IgG3的陽性率明顯高于正常人((2.73±2.09)%vs(0.72±0.42)%,P〈0.05);實驗組GlyCoA+細(xì)胞IgG1的陽性率明顯高于正常人((3.97±2.21)%vs(0.87±0.41)%,P〈0.05);實驗組GlyCoA+細(xì)胞IgG3的陽性率明顯高于正常人((2.24±1.82)%vs(0.71±0.50)%,P〈0.05)。3.應(yīng)用免疫組化方法分別對實驗組,ICUS患者及正常人骨髓有核細(xì)胞進(jìn)行IgG亞型(IgG1,IgG2,IgG3,IgG4)免疫組化染色,11/15名IRH患者計數(shù)100個有核細(xì)胞IgG1陽性細(xì)胞≥3個,在ICUS患者及正常人骨髓有核細(xì)胞涂片染色中IgG1陽性細(xì)胞未見,對上述三組患者骨髓有核細(xì)胞涂片進(jìn)行IgG2,IgG3,IgG4免疫組化染色,陽性細(xì)胞未見。4.應(yīng)用CBA方法對實驗組、IRH恢復(fù)病人、正常人及ICUS患者骨髓上清IgG亞型(IgG1,IgG2,IgG3,IgG4)進(jìn)行檢測。IRH組IgG1(6998.51±2498.38)mg/L明顯高于IRH恢復(fù)組((4702.27±1235.25)mg/L,P〈0.01)正常對照組((5613.20±1640.93)mg/L,P〈0.05)和ICUS組((5166.20±1761.64)mg/L,P〈0.01)。IRH組IgG3(637.63±252.93)mg/L明顯高于IRH恢復(fù)組((415.47±196.60)mg/L,P〈0.01)正常對照組((422.85±212.69)mg/L,P〈0.05)和ICUS組((485.70±197.29)mg/L,P〈0.05)。IgG2四組比較無統(tǒng)計學(xué)意義,IgG4四組比較無統(tǒng)計學(xué)意義。IRH組IgG((10903.29±2576.10)mg/L)明顯高于IRH恢復(fù)組((8468.27±1957.58)mg/L,P〈0.05)。5.實驗組CD5+CD19+/CD19+明顯高于正常對照組及ICUS組((23.34±9.36)%)vs(13.67±5.10)%vs(12.40±3.63)%(P0.01),實驗組CD5+CD19+/CD19+與骨髓上清IgG1水平呈明顯正相關(guān)關(guān)系(r=0.392,P=0.043)。6.實驗組,骨髓上清IgG1水平與中性粒細(xì)胞絕對值呈明顯負(fù)相關(guān)關(guān)系(r=-0.459,P=0.016);骨髓上清IgG3水平與血紅蛋白成明顯負(fù)相關(guān)關(guān)系(r=-0.389,P=0.045)。7.外周血淋巴細(xì)胞IgG1基因相對表達(dá)量在實驗組,恢復(fù)組,正常對照組分別為((5.27±4.83)vs(2.32±2.03)vs(1.58±1.50)),實驗組明顯高于正常對照組(P〈0.05〉,恢復(fù)組與正常對照組比較無統(tǒng)計學(xué)意義。結(jié)論1.IRH患者的骨髓造血細(xì)胞膜上可檢測到IgG抗體。通過流式細(xì)胞學(xué)方法檢測到骨髓造血膜抗體IgG陽性組CD15+及GlyCoA+細(xì)胞膜抗體IgG1及IgG3升高明顯,免疫組化法證實IRH患者骨髓細(xì)胞膜抗體IgG亞型以IgG1為主,提示在IRH中破壞造血的IgG亞型可能為IgG1及IgG3。2.BMMNC-Ab IgG陽性患者骨髓上清IgG1與IgG3升高明顯,IgG1與CD5+CD19+/CD19+比例呈明顯正相關(guān)關(guān)系,IgG1與患者白細(xì)胞呈負(fù)相關(guān)關(guān)系,IgG3與患者血紅蛋白呈明顯負(fù)相關(guān)關(guān)系。通過RT-PCR方法證實IRH患者外周血淋巴細(xì)胞IgG1 mRNA相對表達(dá)量較正常對照組增高。以上提示IRH患者IgG1生成增多,且為血細(xì)胞破壞的主要因素。
[Abstract]:Objective to detect the subtype of antibody immunoglobulin G (immunoglobulin G, IgG) on the hematopoietic cell membrane of Immuno-related Immuno-related hemtocytopenia (IRH) and to detect the content of IgG subtypes in the bone marrow supernatant, and to explore the dominant subtype of IgG in the IRH patients, providing a theory to further reveal the pathogenesis of the IRH patients. Methods 1. subjects were the primary treatment of IRH patients (experimental group), a case control group and a normal control group with unnamed blood cytopenia (ICUS) and normal controls. The following studies were performed: (1) a flow cytometry (FCM) was used to detect the membrane antibodies (IgM, IgG) of IRH patients (CDl5+, CD34+ and GlycoA+ cells); (2) F. CM detection of IgG (+) of IRH patients and normal people (CDl5+ and GlycoA+ cells) membrane antibody (IgG1, IgG3); (3) using immunohistochemical method (IHC) for Ig G (+) IRH patients, ICUS and normal control group marrow nucleated cells to select the bone marrow hematopoietic cell membrane antibody positive initial treatment group as the experimental group. RH remission patients, ICUS initial treatment patients and normal people were respectively cases and normal controls, and the following study was carried out: (1) the content of IgG subtype (IgG1, IgG2, IgG3, IgG4) in bone marrow supernatant (CBA) was detected by flow microsphere capture (CBA); (2) CD5 +CD19+/CD19+ ratio of FCM peripheral blood was detected; (3) RT-PCR method detected peripheral blood lymphatic fine. Results the mRNA expression of cell IgG1 was 58.69% (27/46) in patients with bone marrow hematopoietic cells (27/46) significantly higher than that in case control group 0% and 0% in normal control group. The positive rate of IgM positive rate of hematopoietic cell membrane antibody in bone marrow 80.43% (37/46).2. experimental group was significantly higher than that of normal people (3.13 + 1.80)%vs (0.87 + 0.38)%), P < 0.05); The positive rate of IgG3 in CD15+ cells was significantly higher than that of normal people (2.73 + 2.09)%vs (0.72 + 0.42)%, P < 0.05). The positive rate of IgG1 in GlyCoA+ cells in experimental group was significantly higher than that of normal people (3.97 + 2.21)%vs (0.87 + 0.41)%, P < 0.05), and the positive rate of GlyCoA+ cell IgG3 in experimental group was significantly higher than that of normal people (2.24 + 1.82)%vs (0.71 +%)%, P Immunohistochemistry method was used to immunohistochemical staining of IgG subtypes (IgG1, IgG2, IgG3, IgG4) in the experimental group, ICUS patients and normal human marrow nucleated cells. The 11/15 name IRH patients counted 100 nucleated cell IgG1 positive cells more than 3, and the IgG1 positive cells were not seen in the ICUS patients and normal human bone marrow smear smear staining, and the three groups of patients were observed. Bone marrow smears were smeared for IgG2, IgG3, and IgG4 immunohistochemical staining. The positive cells did not see the.4. application CBA method to the experimental group, IRH recovery patients, normal and ICUS patients with IgG subtype (IgG1, IgG2, IgG3, IgG4) were significantly higher than those of the recovery group (4702.27 + 1235.25) (4702.27 + 1235.25). The normal control group ((5613.20 + 1640.93) mg/L, P < 0.05) and group ICUS (5166.20 + 1761.64) mg/L, P < 0.01).IRH IgG3 (637.63 + 252.93) mg/L were significantly higher than the IRH recovery group (415.47 + 196.60) mg/L, P < 0.01) normal control group (422.85 + 212.69) mg/L. G4 four groups had no statistically significant.IRH group IgG ((10903.29 + 2576.10) mg/L) significantly higher than the IRH recovery group (8468.27 + 1957.58) mg/L, P < 0.05).5. experimental group CD5+CD19+/CD19+ significantly higher than the normal control group and ICUS group (23.34 + 9.36)%) vs (13.67 + 5.10)%vs (12.40 + 3.63)%. The positive correlation (r=0.392, P=0.043).6. experimental group, the IgG1 level of the bone marrow supernatant was negatively correlated with the absolute neutrophils (r=-0.459, P=0.016); the IgG3 level of the bone marrow supernatant was negatively correlated with the hemoglobin (r=-0.389, P=0.045).7. peripheral blood lymphocyte IgG1 gene relative expression in the experimental group, the recovery group, normal. The control group was (5.27 + 4.83) vs (2.32 + 2.03) vs (1.58 + 1.50)), and the experimental group was significantly higher than that of the normal control group (P < 0.05 >). There was no statistical significance between the recovery group and the normal control group. Conclusion the IgG antibody could be detected on the bone marrow hematopoietic cell membrane of the 1.IRH patients. Through the flow cytometry, the IgG positive group of bone marrow hematopoietic membrane antibody CD was detected in CD. The increase of 15+ and GlyCoA+ cell membrane antibodies IgG1 and IgG3 was obvious. The immunohistochemical method confirmed that the IgG subtype of the bone marrow cell membrane antibody of IRH patients was dominated by IgG1, suggesting that the IgG subtype that destroyed the hematopoiesis in IRH was likely to be higher in IgG1 and IgG3.2.BMMNC-Ab IgG positive bone marrow supernatants. There was a negative correlation between IgG1 and leukocyte in patients. IgG3 was negatively correlated with hemoglobin in patients. The relative expression of IgG1 mRNA in peripheral blood lymphocytes of IRH patients was higher than that in normal control group by RT-PCR method. The above suggested that IgG1 production in IRH patients increased and was the main factor of blood cell destruction.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R55

【共引文獻(xiàn)】

相關(guān)博士學(xué)位論文 前4條

1 王一浩;骨髓庫姆試驗（+）血細(xì)胞減少患者骨髓造血細(xì)胞自身抗體IgG破壞或抑制造血的機(jī)制研究[D];天津醫(yī)科大學(xué);2009年

2 劉惠;免疫相關(guān)性全血細(xì)胞減少癥患者骨髓造血細(xì)胞膜靶抗原的初步研究[D];天津醫(yī)科大學(xué);2010年

3 丁凱;免疫相關(guān)性全血細(xì)胞減少癥患者骨髓造血細(xì)胞膜靶抗原的研究[D];天津醫(yī)科大學(xué);2013年

4 任悅;免疫相關(guān)性全血細(xì)胞減少癥患者遺傳易感性的初步研究[D];天津醫(yī)科大學(xué);2014年

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1 于虹;免疫相關(guān)性全血細(xì)胞減少癥患者濾泡輔助性T細(xì)胞數(shù)量及功能研究[D];天津醫(yī)科大學(xué);2011年

2 邴麗娟;骨髓間接Coombs實驗方法的應(yīng)用及其對免疫相關(guān)性血細(xì)胞減少癥患者的臨床診斷意義[D];天津醫(yī)科大學(xué);2012年

3 張江勃;自身免疫性骨髓衰竭癥患者淋巴細(xì)胞和CD34~+細(xì)胞端粒長度研究[D];天津醫(yī)科大學(xué);2014年

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