Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Debate Round Table
  • Published:

Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Geneva experience

Leukemia volume 17, pages 228–231 (2003)Cite this article

Stem cell transplantation (SCT) is an established treatment for various hematological disorders. Usually, the patient's hematopoiesis is ablated by the combination of the conditioning regimen and the allogeneic effect of the graft and the presence of recipient cells in the blood can be a sign of graft failure or of the recurrence of the original disease. Therefore, assays capable of determining the donor or recipient origin may help to assess the success or failure of the transplantation. Information about the quantity of recipient cells in the blood may help to adjust the therapy during first weeks after transplant (graft take) or to diagnose a relapse in patients transplanted for leukemia or myelodysplastic syndrome (MDS). Furthermore, the response of a patient in relapse to a donor lymphocyte infusion can be monitored to decide whether a second dose of allogeneic lymphocytes should be given or not.

In our transplantation unit, all patients transplanted with an allogeneic graft are monitored for the occurrence of mixed chimerism. For patients having received a myelobablative conditioning regimen, the first blood sample is tested between day 15 and 20, followed by a second sample around day 30. Then, when the graft take has occurred, patients are tested at regular intervals (at 2, 3, 6 and 12 months) during the first year and approximately once per year thereafter. Patients having received a non-myeloablative conditioning regimen are monitored more frequently. During the first 2 months after transplantation, they are tested weekly, then every other week until day 100, every other month for up to 1 year and three to four times per year thereafter. Patients treated with DLI are monitored on a weekly basis after the start of the adoptive therapy until full donor chimerism is achieved. In case of suspicion of relapse, chimerism analysis is also part of the diagnostic procedure and is continuously compared to other clinical and molecular markers such a BCR-ABL in patients with recurrent chronic myeloid leukemia (CML). Here, as a contribution to the current debate in Leukemia1,2,3,4,5,6,7,8,9,10 we present our in-house method as applied to 111 stem cell-transplanted patients.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

References

  1. Lion, T & Muller-Berat, N Chimerism testing after allogeneic stem cell transplantation: importance of timing and optimal technique for chimerism testing in different clinical-biological situations. Leukemia, (1999). 13, 2059

    Article  CAS  Google Scholar 

  2. Lion, T Chimerism testing after allogeneic stem cell transplantation: importance of timing and optimal technique for testing in different clinical-biological situations. Leukemia, (2001). 15, 292

    Article  Google Scholar 

  3. Thiede, C, Bornhauser, M, Oelschlagel, U, Brendel, C, Leo, R, Daxberger, H, Mohr, B, Florek, M, Kroschinsky, F, Geissler, G, Naumann, R, Ritter, M, Prange-Krex, G, Lion, T, Neubauer, A & Ehninger, G Sequential monitoring of chimerism and detection of minimal residual disease after allogeneic blood stem cell transplantation (BSCT) using multiplex PCR amplification of short tandem repeat-markers. Leukemia, (2001). 15, 293–302.

    Article  CAS  Google Scholar 

  4. Thiede, C & Lion, T Quantitative analysis of chimerism after allogeneic stem cell transplantation using multiplex PCR amplification of short tandem repeat markers and fluorescence detection. Leukemia, (2001). 15, 303–306.

    Article  Google Scholar 

  5. Lion, T & Muller-Berat, N Chimerism testing after allogeneic stem cell transplantation: importance of timing and optimal technique for chimerism testing in different clinical–biological situations. Leukemia, (2001). 15, 1966

    Article  Google Scholar 

  6. Mattsson, J, Uzunel, M, Tammik, L, Aschan, J & Ringden, O Leukemia lineage-specific chimerism analysis is a sensitive predictor of relapse in patients with acute myeloid leukemia and myelodysplastic syndrome after allogeneic stem cell transplantation. Leukemia, (2001). 15, 1976–1985.

    Article  CAS  Google Scholar 

  7. Gonzalez, M, Lopez-Perez, R, Garcia-Sanz, R, Perez-Simon, JA & San Miguel, JF Debate round-table: comments concerning chimerism studies. Leukemia, (2001). 15, 1986

    Article  CAS  Google Scholar 

  8. Klingebiel, T, Niethammer, D, Dietz, K & Bader, P Progress in chimerism analysis in childhood malignancies – the dilemma of biostatistical considerations and ethical implications. Leukemia, (2001). 15, 1989–1991.

    Article  CAS  Google Scholar 

  9. Biondi, A & Balduzzi, A Questions on chimerism analysis after stem cell transplantation. Leukemia, (2001). 15, 1995

    Article  CAS  Google Scholar 

  10. Lawler, M Prospective chimerism analysis, the time is now but can we respond?. Leukemia, (2001). 15, 1992–1994.

    Article  CAS  Google Scholar 

  11. Jeffreys, AJ, Wilson, V, Neumann, R & Keyte, J Amplification of human minisatellites by the polymerase chain reaction: towards DNA fingerprinting of single cells. Nucleic Acids Res, (1988). 16, 10953–10971.

    Article  CAS  Google Scholar 

  12. Lawler, M, Humphries, P & McCann, SR Evaluation of mixed chimerism by in vitro amplification of dinucleotide repeat sequences using the polymerase chain reaction. Blood, (1991). 77, 2504–2514.

    CAS  PubMed  Google Scholar 

  13. Mackinnon, S, Barnett, L, Bourhis, JH, Black, P, Heller, G & O’Reilly, RJ Myeloid and lymphoid chimerism after T-cell-depleted bone marrow transplantation: evaluation of conditioning regimens using the polymerase chain reaction to amplify human minisatellite regions of genomic DNA. Blood, (1992). 80, 3235–3241.

    CAS  PubMed  Google Scholar 

  14. Roux, E, Helg, C, Chapuis, B, Jeannet, M & Roosnek, E Evolution of mixed chimerism after allogeneic bone marrow transplantation as determined on granulocytes and mononuclear cells by the polymerase chain reaction. Blood, (1992). 79, 2775–2781.

    CAS  PubMed  Google Scholar 

  15. Roux, E, Abdi, K, Speiser, D, Helg, C, Chapuis, B, Jeannet, M & Roosnek, E Characterization of mixed chimerism in patients with chronic myeloid leukemia transplanted with T-cell-depleted bone marrow: involvement of different hematologic lineages before and after relapse. Blood, (1993). 81, 243–248.

    CAS  PubMed  Google Scholar 

  16. Roux, E, Helg, C, Dumont-Girard, F, Chapuis, B, Jeannet, M & Roosnek, E Analysis of T cell repopulation after allogeneic bone marrow transplantation: significant differences between recipients of T cell depleted and unmanipulated grafts. Blood, (1996). 87, 3984–3992.

    CAS  PubMed  Google Scholar 

  17. Polymeropoulos, MH, Rath, DS, Xiao, H & Merril, CR Tetranucleotide repeat polymorphism at the human beta-actin related pseudogene H-beta-Ac-psi-2 (ACTBP2). Nucleic Acids Res, (1992). 20, 1432

    Article  CAS  Google Scholar 

  18. Leclair, B, Fregeau, CJ, Aye, MT & Fourney, RM DNA typing for bone marrow engraftment follow-up after allogeneic transplant: a comparative study of current technologies. Bone Marrow Transplant, (1995). 16, 43–55.

    CAS  Google Scholar 

  19. Dubovsky, J, Daxberger, H, Fritsch, G, Printz, D, Peters, C, Matthes, S, Gadner, H & Lion, T Kinetics of chimerism during the early post-transplant period in pediatric patients with malignant and non-malignant hematologic disorders: implications for timely detection of engraftment, graft failure and rejection. Leukemia, (1999). 13, 2060–2069.

    Article  CAS  Google Scholar 

  20. Sullivan, KM, Mannucci, A, Kimpton, CP & Gill, P A rapid and quantitative DNA sex test: fluorescence-based PCR analysis of X-Y homologous gene amelogenin. Biotechniques, (1993). 15, 636–638, 640–641.

    CAS  PubMed  Google Scholar 

  21. Helg, C, Roux, E, Beris, P, Cabrol, C, Wacker, P, Darbellay, R, Wyss, M, Jeannet, M, Chapuis, B & Roosnek, E Adoptive immunotherapy for recurrent CML after BMT. Bone Marrow Transplant, (1993). 12, 125–129.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by a grant of the Swiss National Science Foundation (No. 3100-65′357.01) and by the ‘Dr Henri Dubois-Ferrière-Dinu Lipatti’ Foundation.

Author information

Authors and Affiliations

  1. Division of Hematology, Department of Internal Medicine, University Hospital, Geneva, Switzerland

    Y Chalandon & B Chapuis

  2. Division of Immunology and Allergology, Department of Internal Medicine, University Hospital, Geneva, Switzerland

    S Vischer & E Roosnek

  3. Division of Oncology, Department of Internal Medicine, University Hospital, Geneva, Switzerland

    C Helg

Authors
  1. Y Chalandon
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S Vischer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C Helg
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B Chapuis
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E Roosnek
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chalandon, Y., Vischer, S., Helg, C. et al. Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Geneva experience. Leukemia 17, 228–231 (2003). https://doi.org/10.1038/sj.leu.2402758

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2402758

Keywords

This article is cited by

Search

Advanced search

Quick links