Ex vivo IFN-gamma secretion by circulating CD8 T lymphocytes: implications of a novel approach for T cell monitoring in infectious and malignant diseases.

Details

Serval ID
serval:BIB_9A06E84F1CD5
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Ex vivo IFN-gamma secretion by circulating CD8 T lymphocytes: implications of a novel approach for T cell monitoring in infectious and malignant diseases.
Journal
Journal of Immunology
Author(s)
Pittet M.J., Zippelius A., Speiser D.E., Assenmacher M., Guillaume P., Valmori D., Liénard D., Lejeune F., Cerottini J.C., Romero P.
ISSN
0022-1767
Publication state
Published
Issued date
06/2001
Peer-reviewed
Oui
Volume
166
Number
12
Pages
7634-7640
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't - Publication Status: ppublish
Abstract
To elucidate the functional heterogeneity of Ag-specific T lymphocyte populations, we combined labeling of lymphocytes with MHC/peptide tetramers and a cell surface affinity matrix for IFN-gamma. Magnetic cell sorting of IFN-gamma-positive lymphocytes allowed the selective enrichment and identification of live Ag-specific cytokine-secreting cells by flow cytometry. Naive, memory, and effector Ag-specific populations were evaluated in healthy HLA-A2 individuals. Significant fractions of influenza- and CMV-specific cells secreted IFN-gamma upon challenge with cognate peptide, consistent with an effector/memory status. The sensitivity of the approach allowed the detection of significant numbers of CMV-specific IFN-gamma-secreting cells ex vivo (i.e., without Ag stimulation). This was not apparent when using previously described assays, namely, ELISPOT or intracellular IFN-gamma staining (cytospot). CD8+ T cells specific for the melamoma-associated Ag Melan-A/MART-1 did not produce IFN-gamma upon challenge with cognate peptide, reminiscent with their naive functional state in healthy individuals. In contrast, CD45RA(low) Melan-A/MART-1 tumor-specific cells from three of three melanoma patients presented levels of activity similar to those found for influenza- or CMV virus-specific lymphocytes, compatible with a functional differentiation into competent effector/memory T lymphocytes in vivo. Notably, a sizable fraction of Melan-A/MART-1-specific cells from a patient secreted IFN-gamma ex vivo following peptide-based vaccination. Thus, the high sensitivity of the assay provides a valuable tool to monitor effector T cell responses in different clinical situations.
Keywords
Antigens, Neoplasm, CD8-Positive T-Lymphocytes/cytology, CD8-Positive T-Lymphocytes/immunology, Cell Differentiation/immunology, Cytomegalovirus/immunology, Enzyme-Linked Immunosorbent Assay, Epitopes, T-Lymphocyte/analysis, Flow Cytometry, HLA-A2 Antigen/analysis, HLA-A2 Antigen/immunology, Humans, Immunomagnetic Separation, Immunophenotyping, Infant, Newborn, Influenza A virus/immunology, Interferon-gamma/blood, Interferon-gamma/secretion, Lymphocyte Count, Melanoma/immunology, Melanoma/pathology, Monitoring, Immunologic/methods, Neoplasm Proteins/analysis, Neoplasm Proteins/immunology, Peptide Fragments/analysis, Peptide Fragments/immunology, Sensitivity and Specificity, Viral Matrix Proteins/analysis, Viral Matrix Proteins/immunology
Pubmed
Web of science
Create date
28/01/2008 11:14
Last modification date
20/08/2019 15:01
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