Induction of immunoglobulin isotype switching in cultured I.29 B lymphoma cells. Characterization of the accompanying rearrangements of heavy chain genes
Department of Molecular Genetics and Microbiology
Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; B-Lymphocytes; Cell Line; Cell Separation; Chromosome Deletion; Cloning, Molecular; DNA; Immunoglobulin A; Immunoglobulin Allotypes; Immunoglobulin E; Immunoglobulin Heavy Chains; Immunoglobulin Idiotypes; Immunoglobulin M; Immunoglobulin alpha-Chains; Immunoglobulin mu-Chains; Immunoglobulins; Lipopolysaccharides; Lymphoma; Mice; Mice, Inbred C57BL; RNA; Recombination, Genetic
Microbiology | Molecular genetics
The murine B cell lymphoma I.29 contains cells expressing surface IgM or IgA with identical heavy chain variable regions (9, 25, and D. Klein and J. Stavnezer, unpublished data). Purified IgM+ cells from the lymphoma have been adapted to culture and induced to switch to IgA, IgE, or IgG2 by treatment with lipopolysaccharide (LPS) or by treatment with a monoclonal anti-I.29 antiidiotype plus LPS. Clones of IgM+ cells have been obtained and induced to switch. Under optimal conditions, 30% of the cells in the culture expressed IgA 8 d after the inducers were added, and by 15 d 90% of the cells were IgA+. In actively switching cultures, up to 50% of the cells whose cytoplasm stained positively with anti-IgA stained simultaneously with anti-IgM, which indicates that the appearance of IgA+ cells in the cultures was due to isotype switching and not to clonal outgrowth. Examination by Southern blotting experiments of the Ig heavy chain genes in I.29 cells before and after switching revealed that isotype switching was accompanied by DNA recombinations that occurred within or immediately 5' to the tandemly repeated switch sequences. Within 3 d after the addition of inducers of switching, the nonexpressed chromosome underwent a variety of deletions or expansions within the S mu region, and a portion of the S alpha regions had undergone a 0.9-kb deletion. In cultures that contained at least 12% IgA+ cells, rearranged, expressed alpha genes, produced by recombination between the S mu region within the expressed mu gene and the S alpha region, were detected.
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Citation: J Exp Med. 1985 Mar 1;161(3):577-601.
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