Simultaneous detection of X and Y chromosomes by two-colour fluorescence in situ hybridization in combination with immunophenotyping of single cells to document chimaerism after sex-mismatched bone marrow transplantation

Bone Marrow Transplantation
M J van TolH J Tanke

Abstract

A powerful approach to documenting engraftment after allogeneic BMT is the quantification of the degree of chimaerism in distinct haematopoietic cell lineages. This cannot be achieved by the recently developed, quantitative, modifications of PCR amplification of highly polymorphic DNA markers, unless this technique is applied to separated cell populations. Here, we report the development of a new method, in which cells are simultaneously characterized by enzymatic immunophenotyping and identified for their origin by two-colour fluorescence in situ hybridization with X and Y chromosome-specific DNA probes (XY-FISH/immunostaining). The method enables the rapid, reliable and quantitative analysis of chimaerism within distinct cell lineages after sex-mismatched BMT, without the requirement for cell separation techniques. This is illustrated by investigation of the pattern of chimaerism in patients receiving a sex-mismatched BMT for the treatment of primary immunodeficiencies. The results obtained with the quantitative XY-FISH/immuno staining method show a good correlation with the data generated by the semi-quantitative analysis of PCR amplified minisatellites in FACS-sorted cell fractions. In addition, XY-FISH/immunostaining was s...Continue Reading

Citations

Dec 12, 2001·British Journal of Haematology·J ScopesF M Gibson
Jun 15, 2011·Current Opinion in Organ Transplantation·James M MathewJoshua Miller
Sep 7, 2006·Cytogenetic and Genome Research·C J YeH H Q Heng
Jun 4, 2004·American Journal of Pharmacogenomics : Genomics-related Research in Drug Development and Clinical Practice·Christian Thiede
May 13, 2005·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·Jorge ReyesThomas E Starzl
Aug 17, 2004·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·George V MazariegosJorge Reyes
May 21, 2016·Pediatric Transplantation·Thomas Green, Jonathan Hind
May 9, 2000·Memórias do Instituto Oswaldo Cruz·J C Voltarelli
Aug 18, 2001·Bone Marrow Transplantation·J M Mathew, J Miller
Feb 15, 2019·Current Opinion in Organ Transplantation·Armando GanozaAjai Khanna

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