Characterization of two human monoclonal antibodies reactive with HLA-B12 and HLA-B60, respectively, raised by in vitro secondary immunization of peripheral blood lymphocytes.
We have developed an in vitro immunization system for the production of B-cell lines that secrete HLA-specific human mAbs. For this purpose, peripheral blood lymphocytes of parous women were stimulated with pools of allogeneic lymphocytes. Preferential outgrowth of B-lymphocytes was effected by inclusion of rIL-2 and a B-cell specific nucleoside analogue. Stimulated B cells were immortalized by EBV transformation, and specific antibody-producing transformants were fused to heteromyeloma or mouse myeloma cell lines, yielding stable hybridomas. This approach has led to the successful development of two human heterohybridomas producing HLA-specific mAbs reactive by complement-mediated cytotoxicity. The specificities of these human mAbs, reactive with HLA-B12(44 + 45) and HLA-B60, respectively, are fully concordant with those of HLA-typing sera.
Department of Immunohaematology and Bloodbank, Leiden University Hospital, The Netherlands.
A
A
Mulder
M
M
Kardol
J
J
Blom
W B
WB
Jolley
C J
CJ
Melief
J W
JW
Bruning
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology*; D002461:Cell Line, Transformed; D005260:Female; D015235:HLA-B Antigens / Q000276:immunology*; D004854:Herpesvirus 4, Human; D006650:Histocompatibility Testing; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007223:Infant; D008297:Male; D051379:Mice; D010298:Parity; D011247:Pregnancy
D000911:Antibodies, Monoclonal; D015235:HLA-B Antigens; C035015:HLA-B12 antigen; C081918:HLA-B60 antigen