Immunoglobulin class switch recombination is impaired in Atm-deficient mice

Joanne M. Lumsden, Thomas McCarty, Lisa K. Petiniot, Rhuna Shen, Carrolee Barlow, Thomas A. Wynn, Herbert C. Morse, Patricia J. Gearhart, Anthony Wynshaw-Boris, Edward E. Max, Richard J. Hodes

Research output: Contribution to journalArticle

Abstract

Immunoglobulin class switch recombination (Ig CSR) involves DNA double strand breaks (DSBs) at recombining switch regions and repair of these breaks by nonhomologous end-joining. Because the protein kinase ataxia telengiectasia (AT) mutated (ATM) plays a critical role in DSB repair and AT patients show abnormalities of Ig isotype expression, we assessed the role of ATM in CSR by examining ATM-deficient mice. In response to T cell-dependent antigen (Ag), Atm-/- mice secreted substantially less Ag-specific IgA, IgG1, IgG2b, and IgG3, and less total IgE than Atm+/+ controls. To determine whether Atm-/- B cells have an intrinsic defect in their ability to undergo CSR, we analyzed in vitro responses of purified B cells. Atm -/- cells secreted substantially less IgA, IgG1, IgG2a, IgG3, and IgE than wild-type (WT) controls in response to stimulation with lipopolysaccharide, CD40 ligand, or anti-IgD plus appropriate cytokines. Molecular analysis of in vitro responses indicated that WT and Atm-/- B cells produced equivalent amounts of germline IgG1 and IgE transcripts, whereas Atm-/- B cells produced markedly reduced productive IgGl and IgE transcripts. The reduction in isotype switching by Atm-/- B cells occurs at the level of genomic DNA recombination as measured by digestion-circularization PCR. Analysis of sequences at CSR sites indicated that there is greater microhomology at the μ-γ1 switch junctions in ATM B cells than in wild-type B cells, suggesting that ATM function affects the need or preference for sequence homology in the CSR process. These findings suggest a role of ATM in DNA DSB recognition and/or repair during CSR.

Original languageEnglish (US)
Pages (from-to)1111-1121
Number of pages11
JournalJournal of Experimental Medicine
Volume200
Issue number9
DOIs
StatePublished - Nov 1 2004
Externally publishedYes

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Keywords

  • Ataxia telangiectasia
  • B lymphocytes
  • DNA damage
  • DNA repair
  • Ig class switching

ASJC Scopus subject areas

  • Immunology

Cite this

Lumsden, J. M., McCarty, T., Petiniot, L. K., Shen, R., Barlow, C., Wynn, T. A., Morse, H. C., Gearhart, P. J., Wynshaw-Boris, A., Max, E. E., & Hodes, R. J. (2004). Immunoglobulin class switch recombination is impaired in Atm-deficient mice. Journal of Experimental Medicine, 200(9), 1111-1121. https://doi.org/10.1084/jem.20041074