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


, : T cell receptor V beta variable gene family expression in human peripheral blood lymphocytes at the mRNA and membrane protein level. Clinical and Experimental Immunology 88(2): 335-340

A polymerase chain reaction (PCR) was developed, using combinations of an oligonucleotide primer for a T cell receptor V.beta. gene family and one for the constant C.beta. gene segments, to assess the expression of each of 20 V.beta. 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 J.beta. oligonucleotide probe. For T cell lines, a signal was observed in the V.beta.8 combination for Jurkat cells, V.beta.5a in HSB cells, V.beta.2 and V.beta.12a in Molt-3 cells and V.beta.2, V.beta.5a and V.beta.12a 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 V.beta. families were detectable. The intensity of the PCR product varied between various V.beta. gene families. Flow cytometric analysis of blood mononuclear cells from the same donors with a restricted series of V.beta. gene family-specific antibodies also revealed the presence of all families. The approach to assess V.beta. gene family expression in heterogeneous populations opens the possibility to study T cell receptor variable gene expression in relation to physiology and pathologic processes.

US$29.90

PMID: 1533355


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