The use of transgenic mice for the production of a human monoclonal antibody specific for human CD69 antigen.
The CD69 antigen is the earliest activation marker expressed on leukocyte surfaces after stimulation and it has been correlated with disease state in a variety of inflammatory and autoimmune diseases. We were interested in the generation of a human monoclonal antibody (mAb) against the CD69 antigen. To do this, mice carrying human Ig transgenes (on an inactivated endogenous immunoglobulin H and Igkappa background) were immunized with rat cells transfected with the human CD69 molecule. From over 2000 hybridoma clones generated in different fusions, we were able to obtain a human monoclonal antibody, hAIM-29, which specifically recognizes human CD69 on the surface of activated-human leukocytes. We demonstrate that the antibody is specific for the human CD69 molecule, as shown by double staining with mouse anti-human CD69 antibodies, ELISA, immunoblot and immunoprecipitation studies. Results of additional experiments show that hAIM-29 activates intracellular calcium influx without Ig cross-linking and enhances phorbol myristate acetate-induced cell proliferation in a manner similar to other mouse anti-CD69 antibodies. This report is the first to describe the isolation and characterization of a novel human mAb, hAIM-29, which may have therapeutic potential in diseases associated with the presence of activated cells expressing CD69 antigen.
Molina
Ana
A
Area de Inmunología, Facultad de Ciencias, Universidad de Vigo, Lagoas-Marcosende s/n 36200 Vigo, Pontevedra, Spain.
Valladares
Mónica
M
Magadán
Susana
S
Sancho
David
D
Viedma
Fernando
F
Sanjuan
Irene
I
Gambón
Francisco
F
Sánchez-Madrid
Francisco
F
González-Fernández
Africa
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D015703:Antigens, CD / Q000032:analysis / Q000276:immunology*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis / Q000276:immunology*; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D006801:Humans; D037181:Lectins, C-Type; D008213:Lymphocyte Activation; D051379:Mice; D008822:Mice, Transgenic; D051381:Rats; D000099192:CD69 Antigens
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D002118:Calcium; D037181:Lectins, C-Type; D000099192:CD69 Antigens