T cell receptor V beta 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 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.
Hu
H Z
HZ
Department of Pathology, University Hospital, Utrecht, The Netherlands.
de Weger
R A
RA
Bosboom-Kalsbeek
K
K
Tilanus
M G
MG
Rozing
J
J
Schuurman
H J
HJ
D001483:Base Sequence; D001782:Blood Donors; D004277:DNA, Single-Stranded; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000097:blood*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology*; D014407:Tumor Cells, Cultured
D004277:DNA, Single-Stranded; D012333:RNA, Messenger; D016693:Receptors, Antigen, T-Cell, alpha-beta