Generation of T Cell Hybridomas From Naturally Occurring FoxP3+ Regulatory T Cells

Methods Mol Biol. 2011;707:39-44. doi: 10.1007/978-1-61737-979-6_3.

Abstract

Generation of regulatory T cells (or Treg) derived hybridomas offers a tool to study their antigen specificity. T cells hybridomas are produced by fusing TCR α-β-thymoma BW5147 with highly dividing T cell population. In vitro anergy of Tregs is an obstacle in generation of highly dividing Treg population for their fusion. In this chapter, we describe a simple and efficient method to generate large number of blasting Treg and their successful fusion with thymoma BW5147. The resultant hybridomas lose Treg-specific transcription factor FoxP3, respond to antigenic stimulation by producing IL-2, and thus allow the evaluation of antigen specific, Tregs-derived TCRs.

MeSH terms

  • Animals
  • Antigens / immunology
  • Forkhead Transcription Factors / metabolism*
  • Hybridomas / cytology*
  • Mice
  • T-Lymphocytes, Regulatory / cytology*
  • T-Lymphocytes, Regulatory / immunology
  • T-Lymphocytes, Regulatory / metabolism
  • Thymoma / metabolism

Substances

  • Antigens
  • Forkhead Transcription Factors
  • Foxp3 protein, mouse