The human T cell receptor V beta repertoire of normal peripheral blood lymphocytes before and after mitogen stimulation. |
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| Authors: | F S Wong M L Hibberd L Wen B A Millward A G Demaine |
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| Affiliation: | Department of Medicine, King''s College School of Medicine and Dentistry, Denmark Hill, London, UK. |
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| Abstract: | Mitogen 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 V beta subgroups, the V beta repertoire of both unstimulated and phytohaemagglutinin (PHA)-stimulated peripheral T cells from eight healthy individuals was investigated. The samples were also analysed by flow cytometry using anti-V beta 2, V beta 5 and V beta 8 MoAbs. A significant increase in the expression of V beta 6, V beta 7.2 and V beta 10.1 was found in all eight samples of PHA-stimulated T cells compared with unstimulated T cells using the PCR method. In contrast, no differences were found between unstimulated and PHA-stimulated T cells by flow cytometry. These results question the validity of using mitogen-stimulated T cells to investigate TCR gene usage. |
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| Keywords: | T cell receptor phytohaemagglutinin flow cytometry polymerase chain reaction |
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