CD4 and CD8 expression and T cell antigen receptor gene rearrangement in early intrathymic precursor cells.
The earliest T precursor population in the adult mouse thymus, considered to have the surface phenotype CD4lo8-3-44+25-Thy-1lo c-kit+ (termed the low CD4 precursor), has been shown to have the capacity to produce B cells and dendritic cells, as well as T cells, and to have the T cell antigen receptor (TCR) C beta gene region in germ-line configuration. Because of evidence that this precursor population may have low levels of CD8 as well as CD4 on the cell surface, it was isolated, stained for surface CD4 and CD8 and assayed for the expression of messenger RNA (mRNA) for CD4 and CD8 by the reverse transcriptase polymerase chain reaction (RT-PCR). The low CD4 precursors gave definite, moderate levels of staining for both CD8 and CD4, in contrast to downstream precursors which showed only marginal staining and so could be considered as genuine CD4-8-3- triple negatives. The low CD4 precursor expressed a significant level of mRNA for CD4, indicating that the surface CD4 was produced by these cells. However, the low CD4 precursor did not express a detectable level of mRNA for CD8, suggesting that the surface CD8 was acquired from other cells. Since the low CD4 precursor population was found already to express mRNA for enzymes involved in TCR gene rearrangement, including in this study terminal deoxynucleotidyl transferase (TdT), a PCR procedure was used to assay early precursors for D-J rearrangements at the TCR beta gene locus. However, the low CD4 precursor had the TCR beta D-J genes in germ-line configuration, D-J gene rearrangements being first detected several stages downstream in the CD3-4-8-44-25-+ precursor population. We conclude that a transient synthesis of CD4, but not of CD8, characterize these early thymus precursors. Although they have initiated synthesis of some recombination-associated enzymes, full commitment to the T lineage and TCR gene rearrangement is a later event.
Ismaili
J
J
Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia.
Antica
M
M
Wu
L
L
D000818:Animals; D001483:Base Sequence; D015704:CD4 Antigens / Q000096:biosynthesis*; D016827:CD8 Antigens / Q000096:biosynthesis*; D002454:Cell Differentiation; D019070:Cell Lineage; D005260:Female; D015329:Gene Rearrangement, T-Lymphocyte; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D016130:Immunophenotyping; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000166:cytology*; D013950:Thymus Gland / Q000166:cytology* / Q000254:growth & development
D015704:CD4 Antigens; D016827:CD8 Antigens