T cell receptor Vβ variable gene family expression in human peripheral blood lymphocytes at the mRNA and membrane protein level.

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 agarose 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 Vβ2 combination for Jurkat cells. Vβ5a in HSB cells, Vβ2 and Vβ2a in Molt-3 cells and Vβ2. Vβ5a and Vβ2a in Moit-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. [ABSTRACT FROM AUTHOR]