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Related Experiment Video

Updated: Jun 12, 2026

Induction and Assessment of Class Switch Recombination in Purified Murine B Cells

Induction and Assessment of Class Switch Recombination in Purified Murine B Cells

Published on: August 13, 2010

p53 represses class switch recombination to IgG2a through its antioxidant function.

Jeroen E J Guikema¹, Carol E Schrader, Michael H Brodsky

¹Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, MA 01655, USA.

Journal of Immunology (Baltimore, Md. : 1950)

|May 21, 2010

Summary

Reactive oxygen species (ROS) are crucial for immunoglobulin class switch recombination (CSR). This study reveals that the antioxidant p53 inhibits CSR by reducing ROS and DNA breaks, thereby regulating B cell immune responses.

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Published on: February 18, 2011

Area of Science:

Immunology
Molecular Biology
Cell Biology

Background:

Immunoglobulin class switch recombination (CSR) is essential for adaptive immunity, diversifying antibody responses.
CSR involves DNA double-strand breaks in switch (S) regions, initiated by activation-induced cytidine deaminase (AID).
Reactive oxygen species (ROS) are implicated as critical intermediates in AID-mediated DNA break formation during CSR.

Purpose of the Study:

To investigate the role of ROS and the p53 protein in regulating CSR.
To determine the impact of antioxidant intervention on CSR.
To elucidate the mechanism by which p53 influences CSR and DNA integrity in B cells.

Main Methods:

Treatment of B cells with N-acetylcysteine (ROS scavenger) and IFN-gamma.
Assessment of CSR induction and IgG2a switching.
Measurement of intracellular ROS levels and p53 activation.
Analysis of DNA breaks and mutations in S regions.

Main Results:

N-acetylcysteine significantly inhibited CSR, confirming the requirement for ROS.
IFN-gamma treatment increased ROS levels and activated p53 in switching B cells.
p53 was found to inhibit IgG2a class switching and suppress DNA breaks and mutations in S regions.
These findings suggest p53 acts as an inhibitor of AID activity.

Conclusions:

ROS are indispensable for CSR, and their scavenging inhibits the process.
p53 plays a critical inhibitory role in CSR, likely by modulating ROS levels and protecting DNA integrity.
p53's regulatory function impacts B cell antibody diversification and genomic stability during CSR.