Generation of a fusion partner to sample the repertoire of splenic B cells destined for apoptosis.
B cells proliferate and diversify in germinal centers in response to antigen. Only a small percentage of these B cells will emerge to form the serum antibody response. Other B cells making lower affinity antibodies, acquiring nonsense mutations, or expressing autoreactivity as a result of somatic mutation undergo an apoptotic cell death and are not efficiently sampled in current analyses of B-cell hybridomas. We have demonstrated that expression of bcl-2 in the NSO myeloma fusion partner leads to a higher yield of viable hybridomas, with a selective increase in hybridomas from B cells that produce autoantibodies and are seldom recovered when spleen cells from non-autoimmune mice are fused to the conventional NSO cell line. Using this fusion partner, we have generated hybridomas from anti-DNA antibody-producing transgenic B cells that are anergic in vivo and destined for apoptosis. These studies provide a strategy to sample the repertoire of B cells that arise in vivo but are not selected to contribute to the expressed antibody response. Furthermore, they demonstrate that restricted expression of bcl-2 in B cells contributes to the maintenance of self-tolerance in secondary lymphoid organs.
Ray
S
S
Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.
Diamond
B
B
D000818:Animals; D017209:Apoptosis; D001402:B-Lymphocytes / Q000166:cytology*; D002459:Cell Fusion; D015870:Gene Expression; D006825:Hybridomas / Q000166:cytology*; D007136:Immunoglobulins / Q000096:biosynthesis; D051379:Mice; D011518:Proto-Oncogene Proteins / Q000502:physiology*; D019253:Proto-Oncogene Proteins c-bcl-2; D011519:Proto-Oncogenes; D012333:RNA, Messenger / Q000235:genetics; D013154:Spleen / Q000166:cytology*; D014162:Transfection
D007136:Immunoglobulins; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger