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PMID:10430621
| Citation |
Schrader, CE, Edelmann, W, Kucherlapati, R and Stavnezer, J (1999) Reduced isotype switching in splenic B cells from mice deficient in mismatch repair enzymes. J. Exp. Med. 190:323-30 |
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| Abstract |
Mice deficient in various mismatch repair (MMR) enzymes were examined to determine whether this repair pathway is involved in antibody class switch recombination. Splenic B cells from mice deficient in Msh2, Mlh1, Pms2, or Mlh1 and Pms2 were stimulated in culture with lipopolysaccharide (LPS) to induce immunoglobulin (Ig)G2b and IgG3, LPS and interleukin (IL)-4 to induce IgG1, or LPS, anti-delta-dextran, IL-4, IL-5, and transforming growth factor (TGF)-beta1 to induce IgA. After 4 d in culture, cells were surface stained for IgM and non-IgM isotypes and analyzed by FACS((R)). B cells from MMR-deficient mice show a 35-75% reduction in isotype switching, depending on the isotype and on the particular MMR enzyme missing. IgG2b is the most affected, reduced by 75% in Mlh1-deficient animals. The switching defect is not due to a lack of maturation of the B cells, as purified IgM(+)IgD(+) B cells show the same reduction. MMR deficiency had no effect on cell proliferation, viability, or apoptosis, as detected by [(3)H]thymidine incorporation and by propidium iodide staining. The reduction in isotype switching was demonstrated to be at the level of DNA recombination by digestion-circularization polymerase chain reaction (DC-PCR). A model of the potential role for MMR enzymes in class switch recombination is presented. |
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| Keywords |
Adaptor Proteins, Signal Transducing; Adenosine Triphosphatases; Animals; B-Lymphocytes/cytology; B-Lymphocytes/enzymology; B-Lymphocytes/metabolism; Base Pair Mismatch/immunology; Carrier Proteins; Cell Cycle/genetics; Cell Cycle/immunology; Cell Division/genetics; Cell Division/immunology; Cell Survival/genetics; Cell Survival/immunology; Cells, Cultured; DNA Repair/immunology; DNA Repair Enzymes; DNA-Binding Proteins; Flow Cytometry; Immunoglobulin Class Switching/genetics; Immunoglobulin Isotypes/biosynthesis; Immunoglobulin Isotypes/genetics; Mice; Neoplasm Proteins/genetics; Neoplasm Proteins/immunology; Nuclear Proteins; Proteins/genetics; Proteins/immunology; Spleen |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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