作者： Hayashi T, Amakishi E, Inoue M, Hirayama F.

来源：Int J Hematol. 2011 Feb 1.

摘要：抗人类血小板抗原（HPAs）抗体在血小板减少症中有着很重要的作用。在日本，HPA-4b抗体常常造成与HPA相关的新生儿同种免疫性血小板减少症。目前已研究出一种用血小板检测抗HPA抗体的高度敏感的检测方法。但是，用于检测的含有表达特异性HPA的血小板难以获得，因此，一种不需要血小板的方法对检测抗HPA抗体非常有帮助。编码HPA-4b糖蛋白IIIa（GPIIIa）的cDNA，利用逆转录病毒载体与野生型GPIIb在K562细胞内进行同源重组，K562细胞属于非粘附细胞系。观察在人工培养细胞中转基因产物的表达6个月以上。下一步评估该细胞株谱细胞的敏感性和特异性。我们用血小板抗原单克隆抗体特异性免疫固定检测技术（MAIPA）检测已用其他方法鉴定过的血清。所有血清标本的HPA-4b抗体都为阳性，而所有对照血清和含抗HLA抗体的血清检测结果为阴性。用我们的细胞株谱细胞和使用传统血小板谱细胞进行MAIPA检测所得的结果在特异性和敏感性上没有差异。目前结果表明，建立的细胞株谱细胞可以高度敏感的检测抗HPA-4b抗体。

Establishment of a cell line panel for the detection of antibodies against human platelet antigen 4b

Hayashi T, Amakishi E, Inoue M, Hirayama F.

Int J Hematol. 2011 Feb 1.

Abstract:Antibodies against human platelet antigens (HPAs) play important roles in thrombocytopenia. In Japan, HPA-4b antibody is frequently responsible for HPA-related neonatal alloimmune thrombocytopenia. A highly sensitive assay using platelets has been developed for the detection of antibodies against HPAs. However, it is difficult to obtain the platelets expressing specific HPAs required for the assay. Therefore, an alternative method not requiring platelets would be helpful to detect antibodies against HPAs. Glycoprotein IIIa (GPIIIa) cDNA encoding HPA-4b was individually co-transduced with that of wild-type GPIIb in K562 cells, which is a non-adherent cell line, using a retroviral vector. The expression of transgene products in cultured cells were observed for over 6 months. Next, to evaluating the sensitivity and specificity of this cell line panel, we performed monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay with a serum previously identified by another method. All HPA-4b antibodies in serum samples were positive, and all serum samples, including normal serum and serum containing HLA antibodies were negative. No difference was observed in the specificity and sensitivity between our method and conventional MAIPA using platelets. The present results indicate that this established cell line panel permits highly sensitive detection of specific antibodies against HPA-4b.

翻译：卢芳 校对：杨亚丽