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Detection of residual host cells after bone marrow transplantation using non-isotopic in situ hybridization and karyotype analysis


, : Detection of residual host cells after bone marrow transplantation using non-isotopic in situ hybridization and karyotype analysis. Bone Marrow Transplantation 11(4): 279-284

Karyotype analysis and interphase cytogenetics by means of non-radioactive in situ hybridization (NISH) with Y and X-specific probes were used to detect residual host cells in BM of 18 male patients who had received a BMT from their sisters. All patients but one had a malignant blood disease; 17 patients were clinically in continuous remission at the time of the investigations and throughout the follow-up period. No host cells were detected by karyotype analysis in patients who were in clinical remission. NISH with a biotinylated Y-specific probe showed residual host cells in 16 of the 17 patients in clinical remission. In samples from three patients the existence of host cells was further confirmed by revealing the XY-containing cells with two-color fluorescence in situ hybridization (FISH) using X- and Y-specific probes simultaneously. Hybridizations with a Y-specific probe on direct BM smears showed gt 3% residual host cells in 50% of the samples studied at or later than 2 months post-BMT. On conventional cytogenetic preparations lt 1% Y-specific cells were detected in all but two samples at gtoreq 2 months post-BMT. There was not difference in the proportion of host cells between patients conditioned with total body irradiation and CY and those who received busulphan and CY. The recipient's stromal or epithelial cells in the aspirates probably account for most of the host cells detected. In conclusion, small numbers of residual host cells detected by interphase cytogenetics with a Y-specific probe do not indicate an imminent relapse. Conventional cytogenetic preparations are preferable to direct smears in studying the presence of residual host hematopoietic cells because of probably less contamination by stromal cells.

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PMID: 8485475


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