title: Establishment of a cell line panel as an alternative source of platelet antigens for a screening assay of anti-human platelet antibodies.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Japanese Red Cross Osaka Blood Center Department of Biochemistry and Molecular Pathology, Medical School, Osaka City University, Osaka, Japan. hayashi-to@osaka.bc.jrc.or.jp
authors.forename: T
authors.initials: T
authors.lastname: Hayashi
authors.forename: E
authors.initials: E
authors.lastname: Amakishi
authors.forename: N
authors.initials: N
authors.lastname: Matsuyama
authors.forename: K
authors.initials: K
authors.lastname: Yasui
authors.forename: R A
authors.initials: RA
authors.lastname: Furuta
authors.forename: Y
authors.initials: Y
authors.lastname: Hori
authors.forename: S
authors.initials: S
authors.lastname: Tanaka
authors.forename: Y
authors.initials: Y
authors.lastname: Fukumori
authors.forename: F
authors.initials: F
authors.lastname: Hirayama
authors.forename: M
authors.initials: M
authors.lastname: Inoue
mesh_terms: D000911:Antibodies, Monoclonal; D016824:Antigens, Human Platelet; D002460:Cell Line; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007518:Isoantibodies / Q000032:analysis* / Q000097:blood
chemical_list: D000911:Antibodies, Monoclonal; D016824:Antigens, Human Platelet; D007518:Isoantibodies