title: Production of human monoclonal anti-Jk3, recognising an epitope including the Jk(a) /Jk(b) polymorphic site of the Kidd glycoprotein.
abstract: OBJECTIVE
  The Kidd blood group system consists of polymorphic antigens, Jk(a) (JK1) and Jk(b) (JK2), and a high-incidence antigen, Jk3. Anti-Jk3 is often observed in immunised Jk(a-b-) individuals. In this study, we aimed to establish a human hybridoma cell line secreting monoclonal anti-Jk3 (HIRO-294).
  
  
  METHODS
  Peripheral blood lymphocytes of a Filipino woman with the Jk(a-b-) phenotype having anti-Jk3 were transformed with Epstein-Barr virus and then hybridised with the myeloma cell line JMS-3 using the polyethylene glycol (PEG) method. The reactivity and specificity of the anti-Jk3 were examined by serology and flow cytometry.
  
  
  RESULTS
  Four hybridoma clones secreting anti-Jk3 were established and the antibody from one of these clones, HIRO-294, was examined. The reactivity of HIRO-294 was positive with 227 Jk(a+b-) red blood cells (RBCs), 298 Jk(a-b+) RBCs, and 1043 Jk(a+b+) RBCs, but was negative with 21 Jk(a-b-) RBCs. Eluates from Jk(a+b-) RBCs and Jk(a-b+) RBCs sensitised with the anti-Jk3 were cross-reacted with Jk(a-b+) RBCs and Jk(a+b-) RBCs, respectively. The reactivity of HIRO-294 was enhanced by the treatment of RBCs with ficin, trypsin, pronase and α-chymotrypsin, but was not changed by their treatment with neuraminidase, dithiothreitol and ethylenediaminetetraacetic acid (EDTA) glycine acid (GA). The RBCs sensitised by the anti-Jk3 were not agglutinated with the commercial reagents of anti-Jk(a) and anti-Jk(b) by saline test, whereas the nonsensitised RBCs or those sensitised by monoclonal anti-D [HIRO-3, immunoglobulin G (IgG) class] were agglutinated with those reagents.
  
  
  CONCLUSIONS
  We established a human hybridoma cell line secreting monoclonal anti-Jk3 (HIRO-294). This antibody had unique specificity, recognising the Kidd glycoprotein including the Jk(a) /Jk(b) polymorphic site.
authors.affiliation: Japanese Red Cross Kanto-Koshinetsu Block Blood Center, Blood Group Section.
authors.forename: C
authors.initials: C
authors.lastname: Toyoda
authors.forename: Y
authors.initials: Y
authors.lastname: Suzuki
authors.forename: H
authors.initials: H
authors.lastname: Tsuneyama
authors.forename: T
authors.initials: T
authors.lastname: Onodera
authors.forename: A
authors.initials: A
authors.lastname: Masuno
authors.forename: R
authors.initials: R
authors.lastname: Yabe
authors.forename: K
authors.initials: K
authors.lastname: Ogasawara
authors.forename: M
authors.initials: M
authors.lastname: Okuda
authors.forename: K
authors.initials: K
authors.lastname: Nakajima
authors.forename: M
authors.initials: M
authors.lastname: Uchikawa
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000918:Antibody Specificity; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D007667:Kidd Blood-Group System / Q000097:blood / Q000235:genetics / Q000276:immunology*; D011110:Polymorphism, Genetic / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D007667:Kidd Blood-Group System
keywords: Jk(a−b−); Kidd blood group; monoclonal anti-Jk3