Real time-PCR assay estimating the naive T-cell pool in whole blood and dried blood spot samples: pilot study in young adults.

Details

Serval ID
serval:BIB_186EE6D0020A
Type
Article: article from journal or magazin.
Collection
Publications
Title
Real time-PCR assay estimating the naive T-cell pool in whole blood and dried blood spot samples: pilot study in young adults.
Journal
Journal of Immunological Methods
Author(s)
Lang P.O., Mitchell W.A., Govind S., Aspinall R.
ISSN
1872-7905 (Electronic)
ISSN-L
0022-1759
Publication state
Published
Issued date
2011
Peer-reviewed
Oui
Volume
369
Number
1-2
Pages
133-140
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Abstract
Because of their central role orchestrating the immune response, the decrease in repertoire number and diversity of naïve T-cells is a significant feature of immnosenescence. Reflecting the effective naive T-cell pool, quantifying the sj-TREC ratio (number of signal joint T-cell receptor excision circles/10(5) T-cells) in blood samples suffers however from constraints. The most limiting one is the absolute requirement of the flow cytometry analysis of peripheral blood samples for the T-cell numeration. In order to make this ratio more accessible for clinical and epidemiological studies addressing how changes in responsiveness of the immune system lead to an increased susceptibility to various diseases and poorer response to vaccination, we have developed a rapid and simple method for the quantification of the sj-TREC ratio in whole blood and in dried blood spot (DBS) samples. This novel method is a QPCR analysis using fluorescently labelled sequence-specific probes both for quantifying sj-TREC and T-cell count and therefore eliminating the absolute necessity of the flow cytometer analysis. In this pilot study, we have compared the sj-TREC ratio we obtained with this novel method in whole blood and in DBS samples of 10 healthy volunteers with those obtained with the technique of reference and found that they are comparable.
Keywords
Adult, Blood Specimen Collection, Humans, Immunity, Innate, Middle Aged, Pilot Projects, Reverse Transcriptase Polymerase Chain Reaction/methods, T-Lymphocytes/immunology
Pubmed
Web of science
Create date
15/04/2015 9:21
Last modification date
20/08/2019 13:48
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