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[骨髓单个核细胞抗人球蛋白试验(+)血细胞减少症患者造血细胞膜自身抗原的初步研究]

[Preliminary study of autoantigens on the membrane of erythropoietic cells of the patients with BMMNC-Coomb's test(+) hemocytopenia].

作者信息

Fu Rong, Liu Hui, Wang Jun, Li Li-juan, Wang Hong-lei, Wang Yi-hao, Shao Zong-hong

机构信息

Department of Hematology, Tianjin Medical University General Hospital, Tianjin 300052, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2012 Oct 16;92(38):2689-93.

Abstract

OBJECTIVE

To observe the relationship between erythropoietin receptor (EPOR) and autoantibodies-IgG/IgM (auto-Ab) on the membrane of erythropoietic cells of the patients with bone marrow mononuclear cells (BMMNC)-Coomb's test(+) hemocytopenia (immunorelated pancytopenia (IRP)) and explore the probable autoantigens of auto-Ab in IRP.

METHODS

A total of 46 newly diagnosed IRP patients (15 with auto-Ab on erythropoietic cells and 31 without) and 18 healthy controls were enrolled. The EPOR expressions on their nuclear erythrocytes were tested with flow cytometry (FCM) to observe the relationship between EPOR and auto-Ab. EPOR mRNA was detected by reverse transcription (RT)-PCR. Stat5 and P-Stat5 proteins in nucleated erythrocytes were measured by Western blot. EPOR expressions on nucleated erythrocytes membrane were re-tested after stripping autoantibodies with glycine buffer.

RESULTS

(1) EPOR of auto-Ab(+) group (1.6% ± 0.9%)was significantly lower than that of auto-Ab(-) group (4.6% ± 4.1%, P < 0.01)and the latter was significantly higher than that of normal controls (2.3% ± 1.8%, P < 0.05). EPOR of IRP patients was inversely correlated with their auto-Ab (r = -0.543, P = 0.000). (2) EPOR mRNA of auto-Ab(+) group (0.68 ± 0.14)was significantly higher than that of auto-Ab(-) group (0.55 ± 0.12, P < 0.01) and normal controls (0.58 ± 0.12, P < 0.05). (3) Protein Stat5 of auto-Ab(+) group (1.45 ± 0.94) was significantly higher than that of normal controls (0.54 ± 0.36, P < 0.05). While P-Stat5 of auto-Ab(+) group (0.42 ± 0.18)was significantly lower than that of normal controls (0.85 ± 0.38, P < 0.05). (4) EPOR expression increased significantly after auto-Ab stripping.

CONCLUSIONS

The auto-Ab of some IRP patients blocks or competitively inhibits EPOR on the membrane of erythropoietic cells. And EPOR may be one of autoantigens in IRP.

摘要

目的

观察骨髓单个核细胞(BMMNC)-抗人球蛋白试验(Coomb's test)阳性血细胞减少症(免疫相关性全血细胞减少症(IRP))患者造血细胞膜上促红细胞生成素受体(EPOR)与自身抗体-IgG/IgM(自身抗体(auto-Ab))之间的关系,探讨IRP中自身抗体的可能自身抗原。

方法

纳入46例新诊断的IRP患者(15例造血细胞存在自身抗体,31例不存在)和18例健康对照者。采用流式细胞术(FCM)检测其有核红细胞上的EPOR表达,以观察EPOR与自身抗体之间的关系。通过逆转录(RT)-PCR检测EPOR mRNA。采用蛋白质免疫印迹法检测有核红细胞中的Stat5和磷酸化Stat5(P-Stat5)蛋白。用甘氨酸缓冲液去除自身抗体后,再次检测有核红细胞膜上的EPOR表达。

结果

(1)自身抗体阳性组的EPOR为(1.6%±0.9%),显著低于自身抗体阴性组(4.6%±4.1%,P<0.01),而自身抗体阴性组显著高于正常对照组(2.3%±1.8%,P<0.05)。IRP患者的EPOR与自身抗体呈负相关(r=-0.543,P=0.000)。(2)自身抗体阳性组的EPOR mRNA为(0.68±0.14),显著高于自身抗体阴性组(0.55±0.12,P<0.01)和正常对照组(0.58±0.12,P<0.05)。(3)自身抗体阳性组的Stat5蛋白为(1.45±0.94),显著高于正常对照组(0.54±0.36,P<0.05)。而自身抗体阳性组的P-Stat5为(0.42±0.18),显著低于正常对照组(0.85±0.38,P<0.05)。(4)去除自身抗体后,EPOR表达显著增加。

结论

部分IRP患者的自身抗体可阻断或竞争性抑制造血细胞膜上的EPOR。且EPOR可能是IRP中的自身抗原之一。

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