特发性血小板减少性紫癜患者脾脏CD_5^+和CD_5^-B细胞共同参与血小板自身抗体的产生  

Participation of both splenic CD_5^+ and CD_5^-B lymphocytes in production of platelet glycoprotein-specific autoantibodies in chronic ITP

作　　者：吕宝君[1] 侯明[1] 芦璐[2] 石艳[1] 何庆泗[1] 马道新[1] 张茂宏[1] 

机构地区：[1]山东大学齐鲁医院,济南250012 [2]山东省交通医院

出　　处：《中华血液学杂志》2002年第9期460-462,共3页Chinese Journal of Hematology

基　　金：山东省科技厅优秀中青年科学家奖励基金资助项目( 9737442 8)

摘　　要：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。Objective To investigate the percentage of splenic CD 5 +B lymphocytes in chronic idiopathic thrombocytopenic purpura (ITP)and the impact of splenic CD 5 + and CD 5 -B lymphocytes on the production of platelet glycoprotein(GP)-specific autoantibodies. Methods Splenic CD 5 +B lymphocytes were identified by two-color flow cytometric analysis in eight patients. Four of the eight patients displayed plasma autoantibodies against both GPⅡb/Ⅲa and GPⅠb/Ⅸ, and their splenic B lymphocytes were separated by Ficoll-Hypaque density gradient and sheep erythrocyte, and further purified by magnetic activate cell separation(MACS). Purified CD 5 + and CD 5 -B lymphocytes were cultured separately with or without staphylococcus aureus cowan I (SAC). GP specific autoantibodies in culture supernatants were measured by modified monoclonal antibody immobilization of platelet antigen assay (MAIPA).Results The percentage of splenic CD 5 +B lymphocytes in ITP patients was slightly higher than that in control with no statistical significance. MACS purified splenic CD 5 + and CD 5 -B lymphocytes from three out of four ITP patients produced high levels of anti-GPⅡb/Ⅲa and anti-GPⅠb/Ⅸ antibodies. Culture supernatants of CD 5 +B lymphocytes from the other patient showed positive reaction only in GPⅠb/Ⅸ MAIPA. Culture supernatant of CD 5 -B lymphocytes from the same patient were double positive in both GPⅡb/Ⅲa and GPⅠb/Ⅸ MAIPA. Conclusions Both splenic CD 5 + and CD 5 -B lymphocytes produce platelet GP-specific autoantibodies in chronic ITP with similar antibody spectrum and titer, and may all play a role in the autoimmune pathogenesis of ITP.

关 键 词：特发性血小板减少性紫癜 自身抗体 脾 B淋巴细胞 CD5抗原 

分 类 号：R554.6[医药卫生—血液循环系统疾病]