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Induction and Assessment of Class Switch Recombination in Purified Murine B Cells
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iNOS potentiates mouse Ig isotype switching through AID expression.

Mi-Ra Lee1, Goo-Young Seo, Young-Myeong Kim

  • 1Department of Molecular Bioscience, College of Biomedical Science, Kangwon National University, Chuncheon 200-701, Republic of Korea.

Biochemical and Biophysical Research Communications
|June 21, 2011
PubMed
Summary

Nitric oxide (NO) is crucial for immunoglobulin A (IgA) synthesis. Inducible nitric oxide synthase deficiency in mice significantly reduces IgA and IgG levels, impacting B cell function and activation-induced cytidine deaminase expression.

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09:49

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Area of Science:

  • Immunology
  • Molecular Biology

Background:

  • Immunoglobulin A (IgA) is vital for mucosal immunity.
  • Nitric oxide (NO) has been identified as a critical factor in mouse IgA synthesis.

Purpose of the Study:

  • To investigate the role of inducible nitric oxide synthase (iNOS) in immunoglobulin (Ig) expression and B cell function.
  • To characterize iNOS-deficient (iNOS(-/-)) mice regarding IgA and IgG production and B cell activation.

Main Methods:

  • Analysis of IgA and IgG levels in fecal pellets, sera, and cultured spleen B cells from iNOS(-/-) and wild-type (WT) mice.
  • Assessment of TGF-β1-inducible IgA and IgG2b synthesis.
  • Evaluation of Ig germ-line transcripts, TGF-β receptor type II (TβRII), BAFF/APRIL, and activation-induced cytidine deaminase (AID) expression.

Main Results:

  • iNOS(-/-) mice exhibited significantly diminished IgA in fecal pellets and reduced IgA production by Peyer's patch cells.
  • Sera and cultured spleen B cells from iNOS(-/-) mice showed substantially lower levels of all IgG subisotypes and IgA.
  • TGF-β1-inducible IgA and IgG2b synthesis was reduced in iNOS(-/-) mice, while Ig germ-line transcripts and TβRII, BAFF/APRIL expression remained comparable.
  • AID expression was diminished in iNOS(-/-) B cells but restored by a NO donor (SNAP).

Conclusions:

  • iNOS plays a critical role in regulating Ig isotype switching, specifically at the level of AID gene expression.
  • NO is essential for maintaining normal levels of IgA and IgG in mice.
  • These findings highlight the importance of the iNOS pathway in adaptive immunity and B cell maturation.