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Updated: Apr 24, 2026

Generation of Human Alloantigen-specific T Cells from Peripheral Blood
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Generation of Human Alloantigen-specific T Cells from Peripheral Blood

Published on: November 21, 2014

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TCR+CD4-CD8- T cells in antigen-specific MHC class I-restricted T-cell responses after allogeneic hematopoietic stem

Raija K Ahmed1, Thomas Poiret, Aditya Ambati

  • 1*Swedish Institute for Infectious Disease Control, SMI †Division of Therapeutic Immunology, Labmed ‡Department of Microbiology, Tumor and Cell Biology ¶Department of Pediatrics, Astrid Lindgen Barnsjukhuset §Center for allogeneic stem cell transplantation (CAST), Karolinska University Hospital, Stockholm, Sweden ∥Department of Immunobiology and Internal Medicine, Yale University, New Haven, CT #Unit of Immunotherapy of Human Tumors, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy.

Journal of Immunotherapy (Hagerstown, Md. : 1997)
|September 9, 2014
PubMed

Human TCRαβ(+) CD4(-)CD8(-) double-negative (DN) T cells represent a minor subset in peripheral blood, yet are important in infectious diseases and autoimmune responses. We examined the frequency of DN T cells in 17 patients after allogeneic hematopoietic stem cell transplantation (aHSCT) at 1, 2, 3, 6, and 12 months post-aHSCT and show that these cells increase early after aHSCT and decrease with time after aHSCT. DN T cells reside in the terminally differentiated effector (CD45RA(+)CCR7(-)) T-cell population and are polyclonal, determined by T-cell receptor Vβ CDR3 analysis. Gene expression analysis of ex vivo sorted DN T cells showed a distinct set of gene expression, including interleukin-8, as compared with CD4(+) or CD8(+) T cells. DN T cells contributed to MHC class I-restricted EBV-directed immune responses, defined by antigen-specific cytokine production and by detection of HLA-A*02:01-restricted EBV BMLF-1 (GLCTLVAML), LMP-2A (CLGGLLTMV), and HLA-A*24:02-restricted EBV BRLF-1 (DYCNVLNKEF) and EBNA3 (RYSIFFDY)-specific T cells. We created retroviral-transfected Jurkat cell lines with a Melan-A/MART-1-specific TCR(+) and the CD8α chain to study TCR(+) DN T cells in response to their nominal MHC class I/peptide ligand. We show that DN T cells exhibit increased TCRζ chain phosphorylation as compared with the TCR(+)CD8(+) transgenic T-cell line. DN T cells contribute to antigen-specific T-cell responses and represent an effector T-cell population that may be explored in immunotherapeutic approaches against viral infections or transformed cells.

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