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Establishment of cell lines stably expressing HNA‐1a, ‐1b, and ‐2a antigen with low background reactivity in flow cytometric analysis

2007; Wiley; Volume: 47; Issue: 3 Linguagem: Inglês

10.1111/j.1537-2995.2006.01139.x

1537-2995

Kazuta Yasui, Tôru Miyazaki, Nobuki Matsuyama, Yoshitaka Kojima, Rika A. Furuta, Jun–ichi Fujisawa, Yoshihiko Tani, Hirotoshi Shibata, Shin‐ichirou Sato, Toshiaki Kato, Hisami Ikeda, Fumiya Hirayama,

Erythrocyte Function and Pathophysiology

Antibodies to neutrophil antigens have been implicated in neonatal alloimmune neutropenia, autoimmune neutropenia, and transfusion-related acute lung injury. Most often, neutrophil-specific antibodies are directed toward human neutrophil antigen (HNA)-1 (Fcgamma receptor 3b) and HNA-2a (CD177) in these disorders.To detect the alloantibodies in the serum samples, a panel of cell lines was established in which the HNA-1a, HNA-1b (polymorphisms of HNA-1), or HNA-2a gene was transduced with a retrovirus vector to confer stable transgene expression in K562 cells that exhibited low background reactivity to human serum samples obtained from healthy donors in flow cytometric analysis.It was shown that several well-characterized human serum samples containing antibodies against HNA-1a, -1b, and -2a were unambiguously identified by the established panel cell lines and observed a lower background reactivity and longer shelf life of the K562 panel cell lines compared with isolated neutrophils, which have been used for the cell panel to identify antibodies against HNA in human serum samples.These results indicate that the K562 panel cell lines provide a good panel for detecting HNA-reactive neutrophil antibodies in human serum samples.