In vitro primary immunization of B lymphocytes for producing human monoclonal antibodies against tumor-associated antigens.
The selective cytotoxicity of the lysosomotropic leucine methyl ester and its lysosomal condensation product leucyl-leucine methyl ester have been used to investigate their effect on a range of lymphocyte subsets and on the cellular proliferation and secretion by immunoglobulin-secreting B cells from axillary regional draining lymph nodes of breast cancer patients. CD2+, CD3+, and CD8+ lymphocyte subsets were selectively reduced by the leucyl-leucine methyl ester treatment (CD2: 84.2-67.5%; CD3: 76.1-62.3%; and CD8: 8.0-3.4%), but there was no significant reduction in the CD4+ and CD19+ subsets (CD4: 68.2-64.7%; and CD19: 22.6-33.2%). In the presence of mouse splenic macrophages as antigen-presenting cells, rIL-2, IFN-gamma, and pokeweed mitogen-stimulated lymphocyte supernatant, leucyl-leucine methyl ester-treated lymphocytes showed a significant increase in 3H-thymidine incorporation and in the number of immunoglobulin-secreting B cells following coculture with the breast tumor cell line MCF-7. In this study, we have characterized some of the cellular and cytokine factors that are necessary for in vitro immunization of human B lymphocytes. Hopefully, this will enable human MAbs to be produced in vitro against tumor-associated antigens.
Li
Y
Y
Department of Surgery, University Medical School, Aberdeen, Scotland, UK.
Ashby
J M
JM
Eremin
O
O
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000912:Antibodies, Neoplasm / Q000276:immunology*; D000938:Antigen-Presenting Cells / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology*; D001365:Axilla; D016175:B-Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology*; D001943:Breast Neoplasms / Q000276:immunology* / Q000473:pathology; D002459:Cell Fusion; D002460:Cell Line; D004151:Dipeptides / Q000494:pharmacology; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008198:Lymph Nodes / Q000473:pathology; D008264:Macrophages / Q000276:immunology; D051379:Mice; D011043:Pokeweed Mitogens / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D004151:Dipeptides; D007376:Interleukin-2; D011043:Pokeweed Mitogens; D011994:Recombinant Proteins; C045139:leucyl-leucine-methyl ester; C030365:leucine methyl ester; D007371:Interferon-gamma; D007930:Leucine