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SUMMARY Background and objectives The Kidd blood group system consists of polymorphic antigens, Jka (JK1) and Jkb (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). Materials and 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-Jka and anti-Jkb 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 Jka/Jkb polymorphic site. |
