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Related Experiment Videos

Clonal evolution of lymphoblastoid cell lines.

Julie L Ryan1, William K Kaufmann, Nancy Raab-Traub

  • 1Department of Dermatology, University of Rochester James P. Wilmot Cancer Center, Rochester, NY, USA.

Laboratory Investigation; a Journal of Technical Methods and Pathology
|October 21, 2006
PubMed
Summary

Epstein-Barr virus (EBV)-immortalized B lymphocytes rapidly become monoclonal in cell culture. These lymphoblastoid cell lines (LCLs) are not ideal controls for normal human B cells due to their single-clone origin.

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Area of Science:

  • Immunology
  • Virology
  • Cell Biology

Background:

  • Lymphoblastoid cell lines (LCLs) are Epstein-Barr virus (EBV)-immortalized B lymphocytes used in research.
  • Understanding LCL clonal evolution is crucial for interpreting experimental results.

Purpose of the Study:

  • To investigate the kinetics of clonal evolution in EBV-infected B lymphocytes.
  • To assess the suitability of LCLs as 'normal' B cell controls.

Main Methods:

  • Tracking immunoglobulin heavy chain (IGH) and T-cell receptor gamma (TRG) gene rearrangements via PCR.
  • Monitoring EBV clonality using Southern blot analysis of viral terminal repeat fragments.
  • Culturing LCLs from five different human donors over time.

Main Results:

Related Experiment Videos

  • All five LCL cultures rapidly evolved to monoclonal B cell populations within 8 weeks.
  • T lymphocytes disappeared from cultures during the observation period.
  • No correlation was found between clonal emergence speed and proliferation rate or EBV fragment size.

Conclusions:

  • EBV-driven B lymphocytes in LCLs undergo rapid clonal expansion, progressing from polyclonal to monoclonal states.
  • Established LCLs do not represent the polyclonal B cell repertoire of healthy individuals.
  • LCLs primarily reflect the progeny of a single B lymphocyte, limiting their use as general 'normal' controls.