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目的 研究慢性特发性血小板减少性紫癜 (ITP)患者脾脏CD5+ B细胞水平的变化及CD5+ 和CD5-B细胞与血小板膜糖蛋白 (GP)特异性自身抗体产生的关系 ,以识别致病B细胞亚群。方法 应用双色流式细胞仪检测 8例慢性ITP患者脾脏CD5+ B细胞水平。选择 4例血浆抗GPⅡb/Ⅲa和抗GPⅠb/Ⅸ抗体双阳性ITP切脾患者 ,应用Ficoll密度梯度离心及花环形成分离法分离脾脏B淋巴细胞 ,继而采用免疫磁珠分选法分选、纯化CD5+ B细胞和CD5-B细胞 ,并分别进行体外培养 ,应用改良MAIPA法检测血浆和细胞培养上清液的血小板特异性抗体。结果 ITP患者脾脏CD5+ B细胞水平较非自身免疫性疾病患者略有增高 ,二者之间差异无统计学意义。CD5+ B细胞水平与患者血小板计数无相关性。 4例血浆抗GPⅡb/Ⅲa和抗GPⅠb/Ⅸ抗体双阳性ITP患者中 ,有 3例CD5+ B细胞、CD5-B细胞培养液中抗GPⅡb/Ⅲa抗体和抗GPⅠb/Ⅸ抗体双阳性。另外 1例CD5+ B细胞培养液抗GPⅡb/Ⅲa抗体阴性 ,抗GPⅠb/Ⅸ抗体阳性 ;CD5-B细胞培养液抗GPⅡb/Ⅲa抗体和抗GPⅠb/Ⅸ抗体双阳性。结论 脾脏CD5+ 和CD5-B细胞均可产生血小板GP特异性自身抗体 ,抗体产生种类和滴度无明显差异。提示二者共同参与了ITP的发病过程
Objective To investigate the changes of splenic CD5 + B cells in patients with idiopathic thrombocytopenic purpura (ITP) and the relationship between CD5 + and CD5-B cells and platelet glycoprotein (GP) specific autoantibodies to identify pathogenic B cells Subgroups. Methods Two-color flow cytometry was used to detect the levels of CD5 + B cells in 8 patients with chronic ITP. Four patients with splenic B lymphocytes from GPIb / Ⅲa and GPⅠb / Ⅸ antibody-positive double-positive ITP splenectomy were selected. The splenic B lymphocytes were isolated by Ficoll density gradient centrifugation and rosette formation and separation. The immunomagnetic beads were used to sort and purify CD5 + B cells and CD5-B cells were cultured in vitro. The platelet-specific antibodies of plasma and cell culture supernatants were detected by modified MAIPA. Results The level of CD5 + B cells in spleen of patients with ITP was slightly higher than that of non-autoimmune diseases, and the difference was not statistically significant. There was no correlation between CD5 + B cell level and patient platelet count. Among 4 cases of plasma anti-GPⅡb / Ⅲa and anti-GPⅠb / Ⅸ antibody double-positive ITP patients, 3 cases of CD5 + B cells, CD5-B cell culture medium anti-GP Ⅱ b / Ⅲ a antibody and anti GP Ⅰ b / Ⅸ antibody double positive. The other 1 CD5 + B cell culture medium was negative for GPⅡb / Ⅲa antibody and positive for GPⅠb / Ⅸ antibody. CD5-B cell culture solution was positive for both GPⅡb / Ⅲa antibody and anti-GPⅠb / Ⅸ antibody. Conclusion Spleen CD5 + and CD5-B cells can produce platelet GP-specific autoantibodies, the type and titer of antibody produced no significant difference. Suggesting that the two involved in the pathogenesis of ITP