EVALUATION OF BCL-2 B CELL TRANSGENIC MICE (B6) FOR HYBRIDOMA PRODUCTION/

Bcl-2 is an oncogene associated with prevention of apoptosis in a vari ety of cell types, Bcl-2 expression in B lymphoid cells prolongs antib ody production, in vitro and in vivo, A line of transgenic mice (B6) h as been developed that expresses human Bcl-2 in the B cells of SWR/SJL mice, B6 transgenic, nontransgenic littermates, and BALB/c mice were immunized with beta-galactosidase (B-gal) or sheep red blood cells (SR BC), The number of spleen cells recovered from immunized B6 mice was 3 -4 times greater than syngeneic, nontransgenic littermates or BALB/c m ice, Spleen cells from B-gal or SRBC immune B6, SWR/SJL, and BALB/c mi ce were fused with P3 myeloma cells to produce hybridomas, Forty-eight percent of the wells plated with fused B6 spleen cells produced B-gal -specific antibodies compared to 14% from BALB/c and 12% from SWR/SJL, Antibody-specific wells were subcloned, resulting in enhanced recover y of antigen-specific subclones with B6-derived fusions compared to co ntrols, In the SRBC fusions, 17% of the wells plated with fused B6 spl een cells produced SRBC-specific antibodies compared to 6% for BALB/c and SWR/SJL spleens, After subcloning, B6-derived clones produced 8% p ositive subclones compared to 9.5% from SWR/SJL and 3.5% from BALB/c, Comparison of the isotype distribution of subclones showed a higher ra tio of IgG antibodies compared to IgM from B6 mice in the B-gal fusion s, IgA antibodies were recovered only from B6 mice, These data indicat e that B6 transgenic mice that overexpress Bcl-2 in their B cells may be superior to other mouse strains for production of antigen-specific hybridomas.