Quantitative and qualitative changes in V-J alpha rearrangements during mouse thymocytes differentiation: implication for a limited T cell receptor alpha chain repertoire.
Knowledge of the complete nucleotide sequence of the mouse TCRAD locus allows an accurate determination V-J rearrangement status. Using multiplex genomic PCR assays and real time PCR analysis, we report a comprehensive and systematic analysis of the V-J recombination of TCR alpha chain in normal mouse thymocytes during development. These respective qualitative and quantitative approaches give rise to four major points describing the control of gene rearrangements. (a) The V-J recombination pattern is not random during ontogeny and generates a limited TCR alpha repertoire; (b) V-J rearrangement control is intrinsic to the thymus; (c) each V gene rearranges to a set of contiguous J segments with a gaussian-like frequency; (d) there are more rearrangements involving V genes at the 3' side than 5' end of V region. Taken together, this reflects a preferential association of V and J gene segments according to their respective positions in the locus, indicating that accessibility of both V and J regions is coordinately regulated, but in different ways. These results provide a new insight into TCR alpha repertoire size and suggest a scenario for V usage during differentiation.
Pasqual
Nicolas
N
Laboratoire d'Immunochimie, Commissariat à l'Energie Atomique, Institut National de la Santé et de la Recherche Médicale, Unité 548, Université Joseph Fourier, 38054 Grenoble Cedex 9, France.
Gallagher
Maighréad
M
Aude-Garcia
Catherine
C
Loiodice
Mélanie
M
Thuderoz
Florence
F
Demongeot
Jacques
J
Ceredig
Rod
R
Marche
Patrice Noël
PN
Jouvin-Marche
Evelyne
E
D000818:Animals; D002454:Cell Differentiation; D015332:Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D013950:Thymus Gland / Q000166:cytology*
D016693:Receptors, Antigen, T-Cell, alpha-beta