TY - JOUR
T1 - Characterization of the T cell receptor repertoire of neonatal T cells by RT-PCR and single strand conformation polymorphism analysis
AU - Alfani, E.
AU - Migliaccio, A. R.
AU - Sanchez, M.
AU - Passarelli, A. M.
AU - Migliaccio, G.
N1 - Funding Information:
This study was supported by institutional funds from Istituto Superiore di Sanità, Rome, Italy, and by contract No. BIO4-CT96–0646 of the European Community. Dr E Mannella, director of the Italian Red Cross in Rome, and Dr H Valenzise, Department of Obstetrician and Gynecology, University of Tor Vergata, Rome, are gratefully acknowledged for providing adult peripheral blood and fetal blood, respectively. Prof G D’Agnolo, chairman of the Laboratory of Cell Biology, is gratefully acknowledged for the continuous support of the study.
PY - 2000
Y1 - 2000
N2 - 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.
AB - 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.
KW - Cord blood
KW - Cord blood transplantation
KW - Graft-versus-host disease
KW - Single strand conformation polymorphism
KW - T cell receptor
UR - http://www.scopus.com/inward/record.url?scp=0033937807&partnerID=8YFLogxK
U2 - 10.1038/sj.bmt.1702434
DO - 10.1038/sj.bmt.1702434
M3 - Article
C2 - 10918409
AN - SCOPUS:0033937807
VL - 26
SP - 83
EP - 89
JO - Bone Marrow Transplantation
JF - Bone Marrow Transplantation
SN - 0268-3369
IS - 1
ER -