具有阵发性睡眠性血红蛋白尿症表型的B淋巴细胞株的建立

Establishment and identification of B cell lines with PNH phenotype

目的建立阵发性睡眠性血红蛋白尿症（PNH）和正常人的B细胞株，用以研究PNH。方法用EB病毒感染PNH患者和正常人的B淋巴细胞，感染后的B淋巴细胞用流式细胞仪分析其细胞表型，然后用免疫磁珠分选CD55-、CD59-B淋巴细胞，并用极限稀释法对分选后的细胞进行克隆，克隆后的细胞株用聚合酶链反应技术扩增免疫球蛋白重链可变区基因重排带，验证其克隆性。结果建立了2株分别来自于2例PNH患者的CD55-、CD59-B细胞株；作为对照，同时建立了2株分别来自于2名正常人的B细胞株。结论可用EB病毒感染B细胞来建立PNH患者的B细胞株。

Objective To establish B cell lines from Patients with paroxysmal nocturnal haemoglobinuria(PNH) and normal controls. Metbods B lymphoid cells prepared from PNH patients and normal controls were in- fected with EB virus. Phenotype of the immortal cells was analyzed by flow cytometry. The immunomagnetic beadswere used to select CD55-and CD59- B lymphoid cells, which were cloned by limiting dilution. The clonality of thecloned cells was confirmed by testing the pattern of Ig remngements using polymerase chain reaction (PCR). Re-sults From 2 PNH patients and 2 normal controls, CD55- and CD59- B lymphoblastoid cell lines were establishedwhich had PNH phenotype and normal phenotype, respectively.Conclusion B lymphoid cell lines from PNH pa-tients can be obtained by EB virus infection.