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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Immunoglobulin Isotypes
Genetic Recombination
B-Lymphocytes
Immunoglobulin G
Immunoglobulin E
Double-Stranded DNA Breaks
Ataxia
Immunoglobulin A
Immunoglobulin Class Switching
Antigens
CD40 Ligand
Sequence Homology
Protein Kinases
Sequence Analysis
Lipopolysaccharides
Digestion
Cytokines
T-Lymphocytes
Polymerase Chain Reaction
DNA

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., ... 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

Immunoglobulin class switch recombination is impaired in Atm-deficient mice. / Lumsden, Joanne M.; McCarty, Thomas; Petiniot, Lisa K.; Shen, Rhuna; Barlow, Carrolee; Wynn, Thomas A.; Morse, Herbert C.; Gearhart, Patricia J.; Wynshaw-Boris, Anthony; Max, Edward E.; Hodes, Richard J.

In: Journal of Experimental Medicine, Vol. 200, No. 9, 01.11.2004, p. 1111-1121.

Research output: Contribution to journalArticle

Lumsden, JM, McCarty, T, Petiniot, LK, Shen, R, Barlow, C, Wynn, TA, Morse, HC, Gearhart, PJ, Wynshaw-Boris, A, Max, EE & Hodes, RJ 2004, 'Immunoglobulin class switch recombination is impaired in Atm-deficient mice', Journal of Experimental Medicine, vol. 200, no. 9, pp. 1111-1121. https://doi.org/10.1084/jem.20041074
Lumsden JM, McCarty T, Petiniot LK, Shen R, Barlow C, Wynn TA et al. Immunoglobulin class switch recombination is impaired in Atm-deficient mice. Journal of Experimental Medicine. 2004 Nov 1;200(9):1111-1121. https://doi.org/10.1084/jem.20041074
Lumsden, Joanne M. ; McCarty, Thomas ; Petiniot, Lisa K. ; Shen, Rhuna ; Barlow, Carrolee ; Wynn, Thomas A. ; Morse, Herbert C. ; Gearhart, Patricia J. ; Wynshaw-Boris, Anthony ; Max, Edward E. ; Hodes, Richard J. / Immunoglobulin class switch recombination is impaired in Atm-deficient mice. In: Journal of Experimental Medicine. 2004 ; Vol. 200, No. 9. pp. 1111-1121.
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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.",
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AU - Wynn, Thomas A.

AU - Morse, Herbert C.

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