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Evaluation of mixed chimerism by in vitro amplification of dinucleotide
repeat sequences using the polymerase chain reaction
M Lawler, P Humphries and SR McCann
Department of Genetics, Trinity College, Dublin, Ireland.
The influence of mixed hematopoietic chimerism (MC) after allogeneic bone
marrow transplantation remains unknown. Increasingly sensitive detection
methods have shown that MC occurs frequently. We report a highly sensitive
novel method to assess MC based on the polymerase chain reaction (PCR).
Simple dinucleotide repeat sequences called microsatellites have been found
to vary in their repeat number between individuals. We use this variation
to type donor-recipient pairs following allogeneic BMT. A panel of seven
microsatellites was used to distinguish between donor and recipient cells
of 32 transplants. Informative microsatellites were subsequently used to
assess MC after BMT in this group of patients. Seventeen of the 32
transplants involved a donor of opposite sex; hence, cytogenetics and Y
chromosome-specific PCR were also used as an index of chimerism in these
patients. MC was detected in bone marrow aspirates and peripheral blood in
18 of 32 patients (56%) by PCR. In several cases, only stored slide
material was available for analysis but PCR of microsatellites or Y
chromosomal material could be used successfully to assess the origin of
cells in this archival material. Cytogenetic analysis was possible in 17
patients and MC was detected in three patients. Twelve patients received
T-cell-depleted marrow and showed a high incidence of MC as revealed by PCR
(greater than 80%). Twenty patients received unmanipulated marrow, and
while the incidence of MC was lower (44%), this was a high percentage when
compared with other studies. Once MC was detected, the percentages of
recipient cells tended to increase. However, in patients exhibiting MC who
subsequently relapsed, this increase was relatively sudden. The overall
level of recipient cells in the group of MC patients who subsequently
relapsed was higher than in those who exhibited stable MC. Thus, while the
occurrence of MC was not indicative of a poor prognosis per se, sudden
increases in the proportions of recipient cells may be a prelude to graft
rejection or relapse.
Volume 77,
Issue 11,
pp. 2504-2514,
06/01/1991
Copyright © 1991 by The American Society of Hematology
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