New insights on human T cell development by quantitative T cell receptor gene rearrangement studies and gene expression profiling.
To gain more insight into initiation and regulation of T cell receptor (TCR) gene rearrangement during human T cell development, we analyzed TCR gene rearrangements by quantitative PCR analysis in nine consecutive T cell developmental stages, including CD34+ lin- cord blood cells as a reference. The same stages were used for gene expression profiling using DNA microarrays. We show that TCR loci rearrange in a highly ordered way (TCRD-TCRG-TCRB-TCRA) and that the initiating Ddelta2-Ddelta3 rearrangement occurs at the most immature CD34+CD38-CD1a- stage. TCRB rearrangement starts at the CD34+CD38+CD1a- stage and complete in-frame TCRB rearrangements were first detected in the immature single positive stage. TCRB rearrangement data together with the PTCRA (pTalpha) expression pattern show that human TCRbeta-selection occurs at the CD34+CD38+CD1a+ stage. By combining the TCR rearrangement data with gene expression data, we identified candidate factors for the initiation/regulation of TCR recombination. Our data demonstrate that a number of key events occur earlier than assumed previously; therefore, human T cell development is much more similar to murine T cell development than reported before.
Dik
Willem A
WA
Department of Immunology, Erasmus MC, 3015 GE Rotterdam, Netherlands.
Pike-Overzet
Karin
K
Weerkamp
Floor
F
de Ridder
Dick
D
de Haas
Edwin F E
EF
Baert
Miranda R M
MR
van der Spek
Peter
P
Koster
Esther E L
EE
Reinders
Marcel J T
MJ
van Dongen
Jacques J M
JJ
Langerak
Anton W
AW
Staal
Frank J T
FJ
D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology*; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000235:genetics / Q000276:immunology*; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor / Q000235:genetics / Q000276:immunology*; D016694:Gene Rearrangement, delta-Chain T-Cell Antigen Receptor / Q000235:genetics / Q000276:immunology*; D019674:Genes, T-Cell Receptor beta / Q000235:genetics / Q000276:immunology; D006801:Humans; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000276:immunology*
D015703:Antigens, CD; D016692:Receptors, Antigen, T-Cell, gamma-delta