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Blood, 15 October 2001, Vol. 98, No. 8, pp. 2456-2465
IMMUNOBIOLOGY
Basic helix-loop-helix proteins E2A and HEB induce immature
T-cell receptor rearrangements in nonlymphoid cells
Anton W. Langerak,
Ingrid
L. M. Wolvers-Tettero,
Ellen J. van
Gastel-Mol,
Monique E. C. M. Oud, and
Jacques J. M. van
Dongen
From the Department of Immunology, Erasmus
University Rotterdam/University Hospital Rotterdam, The
Netherlands.
T-cell receptor (TCR) gene rearrangements are mediated via V(D)J
recombination, which is strictly regulated during lymphoid differentiation, most probably through the action of specific transcription factors. Investigated was whether cotransfection of
RAG1 and RAG2 genes in combination with
lymphoid transcription factors can induce TCR gene
rearrangements in nonlymphoid human cells. Transfection
experiments showed that basic helix-loop-helix transcription
factors E2A and HEB induce rearrangements in the TCRD locus
(D 2-D3 and V2-D3) and TCRG locus ( V 7-J2.3 and V8-J2.3). Analysis of these rearrangements and
their circular excision products revealed some peculiar
characteristics. The V2-D3 rearrangements were formed by direct
coupling without intermediate D2 gene segment usage, and most
D2-D3 recombinations occurred via direct coupling of the
respective upstream and downstream recombination signal sequences
(RSSs) with deletion of the D2 and D3 coding sequences.
Subsequently, the E2A/HEB-induced TCR gene recombination patterns were
compared with those in early thymocytes and acute lymphoblastic
leukemias of T- and B-lineage origin, and it was found that the TCR
rearrangements in the transfectants were early (immature) and not
necessarily T-lineage specific. Apparently, some parts of the
TCRD (V2-D region) and TCRG genes are
accessible for recombination not only in T cells, but also in early
B-cells and even in nonlymphoid cells if the appropriate transcription
factors are present. The transfection system described here appeared to
be useful for studying the accessibility of immunoglobulin and TCR
genes for V(D)J recombination, but might also be applied to study the
induction of RSS-mediated chromosome aberrations.
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