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VH gene replacement in thymocytes.

R Golub1, D Martin, F E Bertrand

  • 1Department of Immunology and Ontario Cancer Institute, University of Toronto, Toronto, Ontario, Canada. rgolub@pasteur.fr

Journal of Immunology (Baltimore, Md. : 1950)
|January 6, 2001
PubMed
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Quasi-monoclonal mice reveal that immunoglobulin heavy chain (IgH) gene replacement can occur in the thymus. This suggests B cell-specific signals or Ig protein are not required for V(H) joining in thymocytes.

Area of Science:

  • Immunology
  • Molecular Biology
  • Genetics

Background:

  • The immunoglobulin heavy chain (IgH) locus undergoes V(D)J recombination, a critical process for adaptive immunity.
  • In normal mice, V(H) gene replacement is largely restricted to B cell development, with thymic rearrangements being rare.
  • The quasi-monoclonal (QM) mouse model allows investigation of IgH locus activities within its natural chromosomal context.

Purpose of the Study:

  • To investigate the rules governing V(D)J recombination at the IgH locus in the thymus.
  • To specifically determine the requirements for V(H) gene replacement in thymocytes.
  • To assess the accessibility and rearrangement potential of IgH genes in the thymus.

Main Methods:

  • Utilizing the quasi-monoclonal (QM) mouse strain with a functionally rearranged H chain gene at the IgH locus.

Related Experiment Videos

  • Analyzing thymocytes, specifically double-positive CD4(+)CD8(+) cells, for evidence of V(H) gene replacement.
  • Cloning and sequencing transcripts from the knocked-in H chain gene.
  • Identifying signal joints associated with V(H) gene replacement.

    • Main Results:

    • Evidence of intermediate products of V(H) replacement was found in QM thymocytes.
    • Transcripts from the knocked-in H chain gene were detected, but no functional mu H chain protein.
    • Sequencing confirmed that some transcripts resulted from V(H) gene replacement events.
    • Corresponding signal joints were identified, indicating successful recombination.

    Conclusions:

    • V(H) gene replacement can occur in thymocytes, challenging previous assumptions.
    • B cell-specific signals or immunoglobulin protein are not essential for activating V(H)-to-VDJ(H) joining in the thymus.
    • Potential mechanisms for overcoming thymic V(H) joining barriers include transcriptionally active gene segments or silencer inactivation.