Antigen triggering selectively increases TCRBV gene transcription.
When the TCR binds Ag it is phosphorylated, internalized, and degraded. We wished to examine whether this process was accompanied by a specific concomitant increase in TCR mRNA levels. To do this, PBMC and a T cell clone were cultured with a series of superantigens and an alloantigen. Only T cells specifically responding to an antigenic stimulus had increased levels of TCR beta-chain variable (TCRBV)-specific mRNA. This response was apparent after 48 h, peaked around 72 h, and was still elevated after 7 days. Increased gene transcription appeared to be driven solely by Ag as specific Ag depletion prevented culture supernatants transferring this effect. The level of TCRBV mRNA elevation was not influenced by the stimulating Ag, but appeared dependent on the gene encoding the stimulated TCR. Reporter gene assays, using cloned TCRBV gene promoters, confirmed both that TCR gene transcription rises after stimulation and that basal and stimulated levels of TCR transcription vary between different TCRBV genes. These data conclusively demonstrate that there is no direct relationship between TCRBV mRNA and T cell number, and that future repertoire studies must take both factors into account.
Department of Molecular and Cellular Pathology, University of Dundee, Ninewells Hospital and Medical School, Dundee, United Kingdom.
G P
GP
Lennon
J E
JE
Sillibourne
E
E
Furrie
M J
MJ
Doherty
R A
RA
Kay
D000328:Adult; D002478:Cells, Cultured; D002999:Clone Cells; D004768:Enterotoxins / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D019674:Genes, T-Cell Receptor beta / Q000276:immunology*; D006801:Humans; D007163:Immunosorbent Techniques; D007424:Intracellular Fluid / Q000276:immunology / Q000378:metabolism; D007519:Isoantigens / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic / Q000276:immunology; D012333:RNA, Messenger / Q000096:biosynthesis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000276:immunology; D018089:Superantigens / Q000276:immunology* / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic / Q000276:immunology*
D004768:Enterotoxins; D007519:Isoantigens; D012333:RNA, Messenger; D018089:Superantigens; C034959:enterotoxin C, staphylococcal