Capture of antigen-specific T lymphocytes from human blood by selective immortalization to establish long-term T-cell lines maintaining primary cell characteristics.
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.
Barsov
Eugene V
EV
AIDS Vaccine Program, SAIC-Frederick, Inc., NCI Frederick, P.O. Box B, Frederick, MD 21702, USA. barsov@ncifcrf.gov
Andersen
Hanne
H
Coalter
Vicky J
VJ
Carrington
Mary
M
Lifson
Jeffrey D
JD
Ott
David E
DE
D000595:Amino Acid Sequence; D000941:Antigens / Q000008:administration & dosage / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002461:Cell Line, Transformed; D049109:Cell Proliferation; D004268:DNA-Binding Proteins / Q000235:genetics; D004168:Diphtheria Toxoid / Q000008:administration & dosage; D016355:Genes, pX; D005822:Genetic Vectors; D006514:Hepatitis B Surface Antigens / Q000008:administration & dosage / Q000235:genetics; D006801:Humans; D007519:Isoantigens; D010446:Peptide Fragments / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D019098:Telomerase / Q000235:genetics; D014161:Transduction, Genetic
D000941:Antigens; D004268:DNA-Binding Proteins; D004168:Diphtheria Toxoid; D006514:Hepatitis B Surface Antigens; D007519:Isoantigens; D010446:Peptide Fragments; D019098:Telomerase