A human monoclonal antibody, produced following in vitro immunization, recognizing an epitope shared by HLA-A2 subtypes and HLA-A28.
In vitro immunization and subsequent immortalization of peripheral blood cells of a multiparous woman has resulted in the production of a stable human mouse heterohybridoma, 5C2A2, secreting an HLA-A2/A28-specific human monoclonal antibody. Although possibly exposed to HLA-A2 by transfusions, the cell donor showed no HLA-A2-specific serum antibodies. The present protocol for in vitro immunization includes the elimination of suppressor cells from the responder cell population, the presence of irradiated allogeneic lymphocytes as a source of antigen, as well as stimuli--recombinant interleukin-2 and a B-cell specific nucleoside analogue--causing the proliferation of B lymphocytes, prior to immortalization. The ability of the antibody 5C2A2 to detect all known HLA-A2 subtypes, except A2.3, and A28, allows identification of the serological epitope on the HLA-A2 molecule. Application of this in vitro immunization method allows the production of a set of HLA monoclonal antibody-secreting human hybridomas, independent of the existence of serum HLA antibodies in the lymphocyte donors.
Mulder
A
A
Department of Immunohematology and Bloodbank, University Hospital Leiden, The Netherlands.
Kardol
M
M
Blom
J
J
Jolley
W B
WB
Melief
C J
CJ
Bruning
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000917:Antibody Formation; D001402:B-Lymphocytes / Q000473:pathology; D002051:Burkitt Lymphoma / Q000276:immunology / Q000473:pathology; D002455:Cell Division; D002461:Cell Line, Transformed; D003601:Cytotoxicity Tests, Immunologic; D000939:Epitopes / Q000276:immunology*; D005260:Female; D015234:HLA-A Antigens / Q000276:immunology*; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000473:pathology; D007114:Immunization; D007525:Isoelectric Focusing; D051379:Mice; D008969:Molecular Sequence Data
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015234:HLA-A Antigens; D015789:HLA-A2 Antigen; C037164:HLA-A28 antigen