Open AccessThe human T cell receptor Vβ repertoire of normal peripheral blood lymphocytes before and after mitogen stimulation
Author(s) -
WONG F. S.,
HIBBERD M. L.,
WEN L.,
MILLWARD B. A.,
DEMAINF A. G.
Publication year - 1993
Publication title -
clinical & experimental immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.329
H-Index - 135
eISSN - 1365-2249
pISSN - 0009-9104
DOI - 10.1111/j.1365-2249.1993.tb03405.x
Subject(s) - phytohaemagglutinin , flow cytometry , t cell receptor , microbiology and biotechnology , biology , stimulation , t cell , t lymphocyte , receptor , beta (programming language) , immunology , antigen , mitogen activated protein kinase , in vitro , endocrinology , immune system , genetics , computer science , programming language
SUMMARY Milogen stimulation of T cells in vitro has been employed in the analysis of the T cell antigen receptor (TCR) repertoire and as a method of generating T cell lines and clones. It has been suspected for some time that mitogen stimulation may bias the repertoire. We have addressed this problem employing a semi‐quantitative technique utilizing the polymerase chain reaction (PCR) and flow cytometry. Using this PCR method and a panel of primers to 22 Vβ subgroups, the Vβ repertoire of both unstimulated and phytohaemagglulinin (PHA)‐stimulated peripheral T cells from eight healthy individuals was investigated. The samples were also analysed by flow cytometry using anti‐Vβ2, Vβ5 and Vβ8 MoAbs. A significant increase in the expression of Vβ6, Vβ7.2 and Vβ 10.1 was found in all eight samples of PHA‐stimulated T cells compared with unstimulatcd T cells using the PCR method. In contrast, no differences were found between unstimulaled and PHA‐stimulated T cells by flow cytometry. These results question the validity of using milogen‐stimulated T cells to investigate TCR gene usage.