Complete TCR-delta rearrangements and partial (D-J) recombination of the TCR-beta locus in CD34+7+ precursors from human cord blood

J Immunol. 1996 Jun 1;156(11):4120-7.

Abstract

In the neonatal human thymus, early immature precursors co-express CD34 and CD7 cell surface Ags, and we have recently shown that its most primitive CD34+7+1- fraction includes TCR-beta-rearranging cells. Bone marrow and cord blood also contain a CD34+7+ population. Although this population is heterogeneous in terms of both phenotype and differentiation capacities, it may include T cell-committed thymus colonizing precursors (prothymocytes). Recently, it has been shown in the mouse that initiation of TCR-beta rearrangements is not restricted to early thymocytes. In the present study, we examined the TCR-beta and TCR-delta rearrangement and transcription status of cord blood CD34+7- and CD34+7+ subpopulations. RNA and DNA were isolated from these cell subsets purified by two consecutive rounds of fluorescence-activated cell sorting from CD3-depleted cord blood cells. Using D-J(beta) or V-J(beta) primer sets and genomic DNA PCR amplification, we showed that CD34+7+, but not CD34+7-, contained D-J(beta) rearrangements without concomitant V-D(beta) recombination. These partial TCR-beta rearrangements within CD34+7+ progenitors were also confirmed at the transcriptional level. Furthermore, whereas none of the CD34+ cord blood cell fraction expressed TCR-alpha transcripts, mature V(delta)1-C(delta) mRNA could be amplified from the CD34+7+ subset. The TdT gene was also transcriptionally active in CD34+7+ cells, thus confirming their lymphoid progenitor content. These data indicate that cord blood CD34+7+ cells, like CD34+7+1- neonatal thymocytes, can initiate TCR-beta gene recombination, reinforcing the idea that T cell commitment may occur before prothymocyte migration to the thymus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD34 / metabolism
  • Antigens, CD7 / metabolism
  • Base Sequence
  • Cell Differentiation
  • DNA Primers / genetics
  • DNA Probes / genetics
  • Fetal Blood / cytology
  • Fetal Blood / immunology*
  • Gene Rearrangement, beta-Chain T-Cell Antigen Receptor
  • Gene Rearrangement, delta-Chain T-Cell Antigen Receptor*
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / immunology*
  • Humans
  • Immunomagnetic Separation
  • Infant, Newborn
  • Mice
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Receptors, Antigen, T-Cell, alpha-beta / genetics*
  • Recombination, Genetic
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology*
  • Transcription, Genetic

Substances

  • Antigens, CD34
  • Antigens, CD7
  • DNA Primers
  • DNA Probes
  • Receptors, Antigen, T-Cell, alpha-beta