Epitopes described in "Quantitation, selection, and functional characterization of Epstein-Barr virus-specific and alloreactive T cells detected by intracellular interferon-gamma production and growth of cytotoxic precursors."

Reference
Article Authors:Guenther Koehne; Katherine M Smith; Teresa L Ferguson; Roxanne Y Williams; Glenn Heller; Eric G Pamer; Bo Dupont; Richard J O'Reilly
Article Title:Quantitation, selection, and functional characterization of Epstein-Barr virus-specific and alloreactive T cells detected by intracellular interferon-gamma production and growth of cytotoxic precursors.
Reference Detail
Reference ID:1004262
Abstract:Techniques for the quantitation of virus-specific and alloantigen-reactive T cells vary in their measurement of clinically relevant T-cell effector populations, their sensitivity and quantitative accuracy, and the time required to obtain measurable results. We compared frequencies of Epstein-Barr virus (EBV)-specific and major alloantigen-reactive T cells as measured by flow cytometric analysis of responding T cells producing intracellular interferon-gamma (IFN-gamma) and by limiting-dilution analysis (LDA) of cytotoxic T-cell precursors (CTLp) at sequential time points during the generation of EBV-specific T-cell lines. The expansion of EBV-specific T lymphocytes and the depletion of alloreactive T cells in cultures of T cells sensitized with autologous EBV-transformed targets followed similar kinetics when measured by either method. Frequencies of EBV- specific T cells generating intracellular IFN-gamma exceeded by 25- to 90-fold the frequencies of responding CTLp at each stage of expansion, whereas the frequencies of alloreactive T cells generating intracellular IFN-gamma exceeded by 30- to 220-fold those detected by LDA. The assay that quantitated T cells producing IFN-gamma yielded more reproducible and precise results than LDA. Furthermore, frequencies detected by the enumeration of T cells responding to immunodominant EBNA 3a and EBNA 3c peptides by IFN-gamma production or their capacity to bind peptide-HLA tetramers were strikingly similar and represented significant fractions of T cells generating IFN-gamma in response to autologous EBV B lymphoblastoid cell line. Functional analysis of responding viable T cells, fractionated on the basis of their secretion of IFN-gamma, demonstrated that EBV-specific and alloantigen cytotoxic T cells were predominantly or exclusively detected in the CD8(+)IFN-gamma(+) fraction of T cells. Strikingly, the CD4(+)IFN-gamma(+) cell fractions were not cytotoxic against EBV-transformed or allogeneic targets.
Affiliations:Department of Pediatrics, Bone Marrow Transplantation Service, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA. koehneg@mskcc.org
Date:2002
Reference Type:Literature
PubMed ID:11861290
Journal:Blood
Journal Volume:99
Article Pages:1730-40
Journal ISSN:1528-0020
Article Chemical List:Biological Markers;Epstein-Barr Virus Nuclear Antigens;Interferon-gamma
Article MeSH List:Biological Markers(analysis); CD4-Positive T-Lymphocytes(immunology; metabolism); CD8-Positive T-Lymphocytes(immunology; metabolism); Cell Separation(methods); Cytotoxicity, Immunologic(immunology); Epstein-Barr Virus Nuclear Antigens(immunology); Flow Cytometry(methods); Herpesvirus 4, Human(immunology); Humans; Immunotherapy, Adoptive(methods); Interferon-gamma(analysis; biosynthesis; diagnostic use); Lymphocyte Count; Lymphocyte Subsets(cytology; immunology); T-Lymphocytes(cytology; immunology; metabolism)
Curation Last Updated:2014-05-14 20:46:47