UMMS Affiliation

Department of Molecular Genetics and Microbiology

Publication Date

1985-03-01

Document Type

Article

Subjects

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

Disciplines

Cells | Genetic Phenomena | Hemic and Lymphatic Diseases | Immune System Diseases | Microbiology | Molecular Genetics | Neoplasms

Abstract

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cells expressing surface IgM or IgA with identical heavy chain variable regions (9, 25, and D. Klein and J. Stavnezer, unpublished data).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.

Rights and Permissions

Publisher PDF posted as allowed by the publisher's terms of use policy at: http://www.rupress.org/terms After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof.

DOI of Published Version

10.1084/jem.161.3.577

Source

J Exp Med. 1985 Mar 1;161(3):577-601.

Journal/Book/Conference Title

The Journal of experimental medicine

Related Resources

Link to article in PubMed

PubMed ID

2579186

Creative Commons License

Creative Commons Attribution-Noncommercial 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 License.

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