The human T cell receptor Vβ repertoire of normal peripheral blood lymphocytes before and after mitogen stimulation

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Abstract
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.