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Related Experiment Videos

Differential usage of VH gene segments is mediated by cis elements

C C Yu1, M Larijani, I N Miljanic

  • 1The Hooper Foundation, San Francisco, CA 94143, USA.

Journal of Immunology (Baltimore, Md. : 1950)
|October 6, 1998
PubMed
Summary

Investigating immunoglobulin (Ig) gene segment recombination, this study used microlocus plasmids to bypass cellular selection. Results show DNA sequence, not cellular activity, drives overuse of specific gene segments like V81x.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Genetics

Background:

  • Immunoglobulin (Ig) diversity is crucial for adaptive immunity.
  • Ig gene diversity arises from V(D)J recombination of gene segments.
  • Developmental restraints and cellular selection influence the final Ig repertoire.

Purpose of the Study:

  • To assess the intrinsic recombination potential of individual Ig gene segments.
  • To investigate Ig gene segment recombination independent of cellular selection and chromatin structure.
  • To determine if DNA sequence alone can explain the preferential usage of certain gene segments.

Main Methods:

  • Construction of Ig heavy (H) chain microlocus plasmids containing germline coding, recombination signal, and flanking sequences of VH, D, and JH segments.

Related Experiment Videos

  • Analysis of recombination events (deletions, additions, and rearrangement types) in the absence of cellular selection.
  • Comparison of plasmid-based rearrangement frequencies with those observed in intact animals.

    • Main Results:

    • Deletions and additions at recombination breakpoints in plasmids mirrored those in vivo.
    • Relative frequencies of different rearrangement types (VD-J, V-DJ, VinvD-J, VDJ) varied significantly.
    • The VH gene segment V81x was overused (15-30x) in the plasmid assay, similar to its usage in intact mice.

    Conclusions:

    • The DNA sequence of Ig gene segments, particularly V81x, can drive their preferential usage in the B cell repertoire.
    • Cellular selection is not solely responsible for the overrepresentation of certain gene segments.
    • Microlocus plasmids provide a valuable tool for studying intrinsic recombination potential.