title: Establishment of a cell line panel for the detection of antibodies against human platelet antigen 4b.
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.
authors.affiliation: Japanese Red Cross Osaka Blood Center, 2-4-43 Morinomiya, Joto-ku, Osaka, 536-8505, Japan. hayashi-to@osaka.bc.jrc.or.jp.|Department of Biochemistry and Molecular Pathology, Medical School, Osaka City University, Osaka, Japan. hayashi-to@osaka.bc.jrc.or.jp.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Hayashi
authors.affiliation: Japanese Red Cross Osaka Blood Center, 2-4-43 Morinomiya, Joto-ku, Osaka, 536-8505, Japan.
authors.forename: Etsuko
authors.initials: E
authors.lastname: Amakishi
authors.affiliation: Department of Biochemistry and Molecular Pathology, Medical School, Osaka City University, Osaka, Japan.
authors.forename: Masayasu
authors.initials: M
authors.lastname: Inoue
authors.affiliation: Japanese Red Cross Osaka Blood Center, 2-4-43 Morinomiya, Joto-ku, Osaka, 536-8505, Japan.
authors.forename: Fumiya
authors.initials: F
authors.lastname: Hirayama
mesh_terms: D016824:Antigens, Human Platelet / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001323:Autoantibodies / Q000097:blood*; D006801:Humans; D039661:Integrin beta3 / Q000235:genetics / Q000378:metabolism; D020014:K562 Cells; D054098:Thrombocytopenia, Neonatal Alloimmune / Q000175:diagnosis* / Q000276:immunology
chemical_list: C510800:4b alloantigen, human; D016824:Antigens, Human Platelet; D001323:Autoantibodies; D039661:Integrin beta3