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Monitoring of cellular chimerism in patients after sex-mismatched bone marrow transplantation: technical report

100%
Jolkowska J., Ladon D., Wachowiak J., Witt M.
Journal of Applied Genetics
|
2000
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vol. 41
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issue 3
209-212
EN
Numerous hematological diseases, in particular leukemias, can be treated successfully with allogeneic bone marrow transplantation (allo-BMT). Highly polymorphic microsatellite markers and X, Y-chromosome-specific sequences provide useful genetic markers for detection of complete or mixed chimerism in patients after allo-BMT. Chimerism can be monitored successfully using polymerase chain reaction technique (PCR) and cytogenetic analysis, especially fluorescent in situ hybridization (FISH). It is still unclear whether individuals with mixed chimerism after bone marrow transplantation have an increased risk of developing leukemic relapse or graft rejection. Molecular study of cellular chimerism can also be used for quantitative assessment of the amount of donor's cells in a recipient after bone marrow transplantation and for monitoring of minimal residual disease (MRD) or disease relapse. We report application of three different DNA-typing techniques: automated DNA sizing technology, fluorescent in situ hybridization and also Y-specific DNA probing for analysis of post-BMT chimerism in a case of sex-mismatched bone marrow transplantation. Key words: allo-BMT, chimerism, FISH, PCR.
2

Implementation of the standard strategy for identification of Ig/TCR targets for minimal residual disease diagnostics in B-cell precursor ALL pediatric patients: Polish experience

76%
Dawidowska M., Jolkowska J., Szczepanski T., Derwich K., Wachowiak J., Witt M.
Archivum Immunologiae et Therapiae Experimentalis
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2008
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vol. 56
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issue 6
409-418
EN
Introduction: Minimal residual disease (MRD), detected based on immunoglobulin and T-cell receptor (Ig/TCR) gene rearrangements as markers of residual leukemic cells, is currently the most reliable prognostic factor in acute lymphoblastic leukemia (ALL). A feasibility study is presented of the standard strategy for the identification of Ig/TCR targets for MRD diagnostics in Polish ALL patients by identifying Ig/TCR gene rearrangement pattern using standard primer sets and protocols. Materials and Methods: The PCR-heteroduplex approach based on BIOMED-1 and BIOMED-2 protocols (recommended as the European standard) was used to detect IGH, IGK-Kde, TCRD, TCRG, and TCRB rearrangements in 58 Polish B-cell precursor ALL patients. Sequencing and homology analysis between the obtained and germline Ig/TCR sequences enabled identification of the rearrangements.The U-Gauss test was used for statistical analysis of the Ig/TCR rearrangement pattern in Polish patients compared with relevant data on other nationalities. Results: The following pattern was identified: IGH: 83% (VH-JH: 74%, DH-JH: 9%), IGK-Kde: 41%, TCRD: 78% (incomplete TCRD: 55%, Vdelta2-Ddelta3: 45%, Ddelta2-Ddelta3: 21%, Vdelta2-Jalpha: 35%), TCRG: 50%, and TCRB: 13%. Considerable convergence of the Ig/TCR pattern in Polish patients and those of other nationalities (mainly West Europeans) was demonstrated. Statistically relevant differences were only found between the incidence of DH-JH in Polish (9%) and Dutch patients (24%; p<0.05) and Polish and Italian patients (19%; p<0.05), VH-JH in Polish (74%) and Chilean patients (100%; p<0.05), and TCRG in Polish (50%) and Brazilian patients (69%; p<0.05). Conclusions: The convergence of Ig/TCR patterns in Polish and European patients indicates that the strategy for Ig/TCR target identification based on standard primers and protocols might be directly used for the construction of Polish standards and recommendations for MRD diagnostics.
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