Characterization of the T cell receptor repertoire of neonatal T cells by RT-PCR and single strand conformation polymorphism analysis

E. Alfani, A. R. Migliaccio, M. Sanchez, A. M. Passarelli, G. Migliaccio

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

We have analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) the individual non-germ line configurations of the T cell receptor (TCR) Vβ chains expressed by T cells from eight individual cord blood specimens. cDNA from each cord blood was amplified using a common primer coupled with a primer specific for each of 22 variable elements of the Vβ chain family and the amplified fragments were separated under high resolution conditions, With cDNA from adult blood (as a control), all of the TCR chains were amplified as a smear consistent with the extensive polyclonality of adult T cells. In contrast, a heterogeneous pattern of amplification was observed with cDNAs from cord blood: only 26.7 ± 21.9% of the 22 Vβ chains analyzed were amplified as a smear. The majority of them were amplified as a discrete number of bands (up to 10) (in 68.2 ± 18.7% of samples) and some of them as a single fragment (4.0 ± 7.8%). Only one of the eight samples analyzed expressed the majority (72.7%) of its Vβ chains as a smear, consistent with an adult-like TCR repertoire. In conclusion, cord blood expressed, on average, a less complex TCR repertoire than adult blood.

Original languageEnglish
Pages (from-to)83-89
Number of pages7
JournalBone Marrow Transplantation
Volume26
Issue number1
DOIs
StatePublished - 2000
Externally publishedYes

Keywords

  • Cord blood
  • Cord blood transplantation
  • Graft-versus-host disease
  • Single strand conformation polymorphism
  • T cell receptor

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