Methods for the ex vivo characterization of human CD8+ T subsets based on gene expression and replicative history analysis.

Details

Serval ID
serval:BIB_3E9FE2D4943E
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Methods for the ex vivo characterization of human CD8+ T subsets based on gene expression and replicative history analysis.
Journal
Methods in molecular medicine
Author(s)
Rufer N., Reichenbach P., Romero P.
ISSN
1543-1894
Publication state
Published
Issued date
2005
Volume
109
Pages
265-84
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't - Publication Status: ppublish
Abstract
The generation of an antigen-specific T-lymphocyte response is a complex multi-step process. Upon T-cell receptor-mediated recognition of antigen presented by activated dendritic cells, naive T-lymphocytes enter a program of proliferation and differentiation, during the course of which they acquire effector functions and may ultimately become memory T-cells. A major goal of modern immunology is to precisely identify and characterize effector and memory T-cell subpopulations that may be most efficient in disease protection. Sensitive methods are required to address these questions in exceedingly low numbers of antigen-specific lymphocytes recovered from clinical samples, and not manipulated in vitro. We have developed new techniques to dissect immune responses against viral or tumor antigens. These allow the isolation of various subsets of antigen-specific T-cells (with major histocompatibility complex [MHC]-peptide multimers and five-color FACS sorting) and the monitoring of gene expression in individual cells (by five-cell reverse transcription-polymerase chain reaction [RT-PCR]). We can also follow their proliferative life history by flow-fluorescence in situ hybridization (FISH) analysis of average telomere length. Recently, using these tools, we have identified subpopulations of CD8+ T-lymphocytes with distinct proliferative history and partial effector-like properties. Our data suggest that these subsets descend from recently activated T-cells and are committed to become differentiated effector T-lymphocytes.
Keywords
CD8-Positive T-Lymphocytes, Cell Division, Fibroblasts, Flow Cytometry, Gene Expression, Humans, In Situ Hybridization, Fluorescence, Reverse Transcriptase Polymerase Chain Reaction, T-Lymphocyte Subsets, Telomere
Pubmed
Create date
28/01/2008 11:28
Last modification date
20/08/2019 13:35
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