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Southern blot patterns, frequencies, and junctional diversity of T-cell
receptor-delta gene rearrangements in acute lymphoblastic leukemia
TM Breit, IL Wolvers-Tettero, A Beishuizen, MA Verhoeven, ER van Wering and JJ van Dongen
Department of Immunology, Erasmus University/University Hospital Dijkzigt,
Rotterdam, The Netherlands.
Southern blot analysis of T-cell receptor (TCR)-delta gene rearrangements
is useful for diagnostic studies on the clonality of lymphoproliferative
diseases. We have developed 18 new TCR-delta gene probes by use of the
polymerase chain reaction (PCR) techniques. Application of these probes for
detailed analysis of the TCR-delta genes in normal control samples, 138
T-cell acute lymphoblastic leukemias (T-ALL), and 91 precursor B-ALL
allowed us to determine the TCR-delta gene restriction map for five
restriction enzymes, as well as the Southern blot restriction enzyme
patterns of all theoretically possible TCR-delta gene rearrangements. Based
on this information, it appeared that 97% of all 213 detected TCR-delta
gene rearrangements in our series of ALL could be detected by use of the
TCRDJ1 probe and that the majority (76%) of the 213 rearrangements could be
identified precisely. In T-ALL, we found a strong preference for the
complete rearrangements V delta 1-J delta 1 (33%), V delta 2-J delta 1
(10%), and V delta 3-J delta 1 (7%) and the incomplete rearrangement D
delta 2- J delta 1 (11%). In precursor B-ALL, the majority of
rearrangements consisted of V delta 2-D delta 3 (72%) and D delta 2-D delta
3 (10%). The junctional diversity of these 6 preferential TCR-delta
rearrangements was analyzed and showed an extensive junctional insertion
(approximately 30 nucleotides) for complete V delta-J delta rearrangements,
whereas incomplete rearrangements had correspondingly smaller junctional
regions. The detailed TCR-delta gene restriction map and probes presented
here, in combination with the Southern blot patterns of TCR-delta gene
rearrangements, are important for TCR-delta gene studies in ALL; all
TCR-delta gene rearrangements can be detected and the majority can be
identified precisely. Identification of rearrangements is a prerequisite
for subsequent PCR analysis of TCR- delta gene junctional regions, eg, for
detection of minimal residual disease during follow-up of ALL patients.
Volume 82,
Issue 10,
pp. 3063-3074,
11/15/1993
Copyright © 1993 by The American Society of Hematology
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