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Molecular analysis of T-cell receptor repertoire in bone marrow transplant
recipients: evidence for oligoclonal T-cell expansion in graft-versus-host
disease lesions
X Liu, V Chesnokova, SJ Forman and DJ Diamond
Department of Hematology and Bone Marrow Transplantation, City of Hope
Medical Center, Duarte, CA 91010, USA.
We have analyzed the T-cell receptor (TCR) V beta repertoire using
polymerase chain reaction (PCR) in a cohort of eight patients receiving
allogeneic bone marrow transplantation (BMT) from related and unrelated
donors at the City of Hope. Results of PCR studies from graft-versus- host
disease (GVHD) skin lesions show a bias in the usage of TCR V beta
families, whereas examination of peripheral blood (PB) withdrawn at the
same time did not reveal a similar phenomenon. In one such family, TCR V
beta 2 is predominantly expressed in 7 of 7 biopsy specimens examined. V
beta 2 TCR expression from these patients was analyzed more extensively
using a combination of individual TCR gene cloning, followed by sequence
analysis. We found evidence of oligoclonal expansion of single V beta
2-bearing TCRs in GVHD lesions, and in the PB of some patients after
diagnosis of GVHD. In contrast, GVHD-negative biopsy samples showed no
evidence for clonotypic TCR amplification. Sequence-specific TCR CDR3
region probes were derived from analysis of the predominant expressed TCR
in GVHD lesions, and used to probe Southern blots of amplified V beta 2 TCR
mRNA from PB and tissue from BMT recipients and their respective donors. In
most cases the probes are highly specific in detecting TCR expression from
GVHD lesions alone, although in several instances expression could be
detected in PB after GVHD diagnosis. These data provide supporting evidence
for the hypothesis that acute GVHD is associated with expansion of T-cell
clones expressing antigen-specific TCRs that may contribute to the disease
pathology.
Volume 87,
Issue 7,
pp. 3032-3044,
04/01/1996
Copyright © 1996 by The American Society of Hematology
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