作者：Hayashi T, Amakishi E, Matsuyama N, Yasui K, Furuta RA, Hori Y, Tanaka S, Fukumori Y, Hirayama F, Inoue M.来源：Transfus Med. 2011 Jan 6. 摘要 背景：由于一些人类血小板抗原（HPAs）的表达比较罕见，所以难以获得用于血小板抗体筛查检测所需的表达各种人类血小板抗原的血小板谱。因此，一个不使用血小板的替代方法将有助于HPA抗体的检测。本研究描述了能稳定表达特异性HPAs的细胞系的建立及它们在血小板特异性抗体检测中的应用。方法：野生型β3，HPA-1b, -6b, -7b 和-7变异体的cDNA以及野生型αIIb 和 HPA-3b的cDNA被分别的与野生型αIIb和β3的cDNA共同转染K562细胞系。我们使用MAIPA方法评估该谱细胞系在血小板抗体检测中的效果，实验中所使用的血清均为经混合被动凝集试验证实含有HPA抗体的血清。结果和结论：在含有HPA-1a (n = 2)、HPA-3a (n = 6)、 HPA-6b (n = 3) 或 HPA-7 变异体(n = 1)抗体的12个血清中，使用我们的新方法检测，除2例HPA-3a抗体未检出外，其余所有的抗体均被检测出。未被检出的2例HPA-3a抗体血清中的一个血清在常规的血小板MAIPA实验中也得到阴性结果，这说明该谱细胞系可以作为血小板抗原的替代来源应用于MAIPA实验中。 Establishment of a cell line panel as an alternative source of platelet antigens for a screening assay of anti-human platelet antibodies Hayashi T, Amakishi E, Matsuyama N, Yasui K, Furuta RA, Hori Y, Tanaka S, Fukumori Y, Hirayama F, Inoue M.Transfus Med. 2011 Jan 6. Abstract Background: A panel of platelets expressing various human platelet antigens (HPAs) for a platelet antibody screening assay is difficult to prepare because some antigens are rarely expressed. Therefore, an alternative method without using platelets would be helpful in detecting HPA antibodies. This study describes the establishment of cell lines that stably express specific HPAs and their application for detecting specific antibodies.Methods: Wild-type β3, HPA-1b, -6b, -7b and -7 variant cDNA as well as wild-type αIIb and HPA-3b cDNA were individually co-transduced with wild-type αIIb and β3 cDNA in the K562 cell line. We performed an immunobead monoclonal antibody immobilisation of platelet antigens (MAIPA) assay to evaluate this cell line panel for antibody detection using identified sera containing HPA antibodies, whose specificities had been determined by the mixed passive haemagglutination test.Results and Conclusion: Of the 12 sera containing HPA-1a (n = 2), HPA-3a (n = 6), HPA-6b (n = 3) or HPA-7 variant (n = 1) antibodies, all antibodies were detected and determined by our new method, except for two HPA-3a antibodies. One of the two antibodies was also negative for conventional platelet MAIPA, suggesting that the cell line panel might be used as an alternative source of platelet antigens in the MAIPA assay.

                                                                      翻译：李丽兰 校对：黎海燕