Capture of antigen-specific T lymphocytes from human blood by selective immortalization to establish long-term T-cell lines maintaining primary cell characteristics

Immunol Lett. 2006 May 15;105(1):26-37. doi: 10.1016/j.imlet.2005.11.028. Epub 2005 Dec 21.

Abstract

To establish long-term, antigen-specific T-cell lines and clones, we selectively immortalized antigen-responsive T cells from human peripheral blood mononuclear cells (PBMCs). PBMCs were stimulated with either alloantigen or soluble antigen, then infected with a murine leukemia virus-based retroviral vector carrying an immortalizing gene, either the Tax gene from human T-cell leukemia virus type 1, or the human telomerase-reverse transcriptase gene. Since such vectors can only integrate in dividing cells, only antigen-activated T cells are efficiently transduced. This approach generated immortalized antigen-specific CD4+ and CD8+ T-cell lines that maintained strictly IL-2-dependent growth and HLA-restricted, antigen-specific responsiveness, some of which have been in continuous culture for longer than 1 year, far in excess of the survival of parallel control non-immortalized cultures. Clones derived from these lines showed antigen-specific proliferation with induced cytokine and chemokine production, and, in the case of a CD8+ T-cell clone, antigen-specific cytolytic activity. This approach provides a convenient, reproducible means for generating a stable, continuously renewable source of antigen-specific T lymphocytes for a variety of studies of T cell biology.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Antigens / administration & dosage
  • Antigens / genetics
  • Cell Culture Techniques / methods*
  • Cell Line
  • Cell Line, Transformed
  • Cell Proliferation
  • DNA-Binding Proteins / genetics
  • Diphtheria Toxoid / administration & dosage
  • Genes, pX
  • Genetic Vectors
  • Hepatitis B Surface Antigens / administration & dosage
  • Hepatitis B Surface Antigens / genetics
  • Humans
  • Isoantigens
  • Peptide Fragments / administration & dosage
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology*
  • Telomerase / genetics
  • Transduction, Genetic

Substances

  • Antigens
  • DNA-Binding Proteins
  • Diphtheria Toxoid
  • Hepatitis B Surface Antigens
  • Isoantigens
  • Peptide Fragments
  • Telomerase