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KMID : 0350519950480041003
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Journal of Catholic Medical College
1995 Volume.48 No. 4 p.1003 ~ p.1015
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Analysis of Engraftment using in Vitro Amplification of Mini-or Mierosatellite Sequences after Allogeneic Bone Marrow Transplantation
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Abstract
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Erythrocyte antigen typing, karyotyping and molecular genetic analysis have been used to document successful and long-term engraftment after allogeneic bone marrow transplantation. Since all these methods are not sensitive enough to allow
detection
of
small numbers of host or donor cells, it has been difficult to assess presence of the donor cells at an early post-transplant or graft failure stage.
Recently, in vitro amplification of mini- or microsatellites loci using polymerase chain reaction (PCR) has been introduced which allows the analysis at very small numbers of host or donor cells in post-transplant state.
In this study, YNZ 22 minisatellite, APO C2 microsatellite and Y-microsatellite were used to assess early engraftment and graft failure on 33 patients with allogeneic bone marrow transplantation.
We determined the sensitivity of this methods for detecting donor specific alleles with mixing experiments, which demonstrated as ranging from 2.4% ~5.5% (YNZ 22) and 0.01% (Y-microsatellite).
Theis sensitivity seemed to be almost as good as polymerase chain reactin-fragment length polymorphism (PCR-FLP) analysis.
Variable number of tandem repeats (VNTR) PCR analysis using YNZ 22 and APO C2 primer was able to confirm engraftment in all patients investigated as early as 14 days after BMT from 10-20 ml peripheral blood. The incidence of mixed chimerism was
30%
(3/10 cases) when tested later than 3 months after transplantation.
Also, we analysed the origin of peripheral blood leukocytes in 9 patients with persistent pancytopenia. 3 patients grafted for leukemia were found to be in microscopic and clinical relapse, although VNTR PCR analysis at this event had shown only
donor
bands. Thus, VNTR PCR analysis seems of little help to predict relapse in our study . In the other patient in whom host cells were not detectable the pancytopenia and marrow hypoplasia were associated with chronic graft-versus-host disease. In 5
patients with rejection, 4 patients had only recipient derived peripheral blood cells. These results have shown the facts that persistence of residual host cells was associated with a high risk of graft rejection. In the other patient with
rejection,
mixed chimerism was detected at 2 and 4 months.
Finally, this study shows that the analysis of mini- and microsatellite VNTR loci by PCR seems to be sensitive method to analyze the origin of hematopoietic cells after allogeneic bone marrow transplantation.
Such analysis performed during early stage and post-transplant pancytopenia will provide important informations into assessment of engraftment.
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