Open AccessT cell receptor Vβ variable gene family expression in human peripheral blood lymphocytes at the mRNA and membrane protein level
Author(s) -
HU H. Z.,
WEGER R. A.,
BOSBOOMKALSBEEK K.,
TILANUS M. G. J.,
ROZING J.,
SCHUURMAN H. J.
Publication year - 1992
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.1365-2249.1992.tb03083.x
Subject(s) - immunology , messenger rna , gene , gene expression , biology , receptor , peripheral blood , cell surface receptor , cell , microbiology and biotechnology , genetics
SUMMARY A polymerase chain reaction (PCR) was developed, using combinations of an oligonucleotide primer for a T cell receptor Vβ gene family and one for the constant Cβ gene segments, to assess the expression of each of 20 Vβ gene families in RNA after reverse transcription into cDNA. The detection was done after agarosc gel electrophoresis of PCR products and ethidium bromide staining. The positive identification of the PCR products was done by hybridization with a Jβ oligonucleotide probe. For T cell lines, a signal was observed in the Vβ8 combination for Jurkat cells, Vβ 5a in HSB cells, Vβ 2 and Vβ12a in Molt‐3 cells and Vβ 2, Vβ 5a and Vβ12a in Molt‐4 cells. Using mixtures of RNA from different cell lines, the sensitivity of the method was in the range of 0·1–0·5%. In peripheral blood mononuclear cells from four donors, taken at three different occasions, all Vβ families were detectable. The intensity of the PCR product varied between various Vβ gene families. Flow cytometric analysis of blood mononuclear cells from the same donors with a restricted series of Vβ gene family‐specific antibodies also revealed the presence of all families. The approach to assess Vβ gene family expression in heterogeneous populations opens the possibility to study T cell receptor variable gene expression in relation to physiology and pathologic processes.