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.

  • Technical Report
  • Published:

Purging in BCR-ABL-positive acute lymphoblastic leukemia using immunomagnetic beads: comparison of residual leukemia and purging efficiency in bone marrow vs peripheral blood stem cells by semiquantitative polymerase chain reaction

Bone Marrow Transplantation volume 25, pages 97–104 (2000)Cite this article

Abstract

Twenty autologous bone marrow (BM) and 25 peripheral blood stem cell (PBSC) grafts were collected from a total of 40 consecutive patients with BCR-ABL+ acute lymphoblastic leukemia (ALL) in first (n = 37) or second (n = 3) complete morphological remission and subsequently purged with a cocktail of anti-CD19, -CD10, AB4 MoAbs and immunomagnetic beads (IMB). Residual BCR-ABL-positive cells before purging were detected in 19 of 20 BM grafts at a median of 4 (range 0–6) logs and in 17 of 25 evaluable PBSC grafts at a median of 1 (range 0–3) log above the limit of detection assessed by a semiquantitative limiting log10-dilution RT-PCR (P < 0.0001). imb purging depleted a median of 2.5 (range 1–4) log of residual bcr-abl+ cells from BM and a median of 1 (range 0–2) log from PBSC grafts, achieving RT-PCR negativity in 1/20 BM and 12/25 PBSC grafts after purging. Cell recoveries were 62% and 86% (P < 0.0001) of mnc and 74% and 97% (P = 0.065) of CD34+ cells after BM and PBSC purging, respectively. BM purging was superior using the triple MoAb cocktail which depleted 2.64 ± 0.4 log (n = 14) compared to 1.6 ± 0.4 log (n = 5) using the MoAb cocktail not including AB4 (P = 0.02). We conclude that unpurged BM grafts contain 2–3 log more residual BCR-ABL+ cells than unpurged PBSC grafts and that purging efficacy is superior in BM compared to PBSC grafts, but median titers in purged BM grafts still exceed those in purged PBSC grafts. Bone Marrow Transplantation (2000) 25, 97–104.

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

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  1. Bloomfield CD, Goldman AI, Alimena G et al. Chromosomal abnormalities identify high-risk and low-risk patients with acute lymphoblastic leukemia Blood 1986 67: 415–420

    CAS  PubMed  Google Scholar 

  2. Secker-Walker LM, Craig JM, Hawkins JM, Hoffbrand AV . Philadelphia positive acute lymphoblastic leukemia in adults: age distribution, bcr breakpoint and prognostic significance Leukemia 1990 5: 196–199

    Google Scholar 

  3. Maurer J, Jannsen JWG, Thiel E et al. Detection of chimeric BCR-ABL genes in acute lymphoblastic leukemia by the polymerase chain reaction Lancet 1991 337: 1055–1058

    Article  CAS  PubMed  Google Scholar 

  4. Hoelzer D . Therapy of the newly diagnosed adult with acute lymphoblastic leukemia Hematol Oncol Clin North Am 1993 7: 139–160

    Article  CAS  PubMed  Google Scholar 

  5. Hoelzer D . Treatment of acute lymphoblastic leukemia Semin Hematol 1994 31: 1–15

    CAS  PubMed  Google Scholar 

  6. Barrett AJ, Horowitz MM, Ash RC et al. Bone marrow transplantation for Philadelphia chromosome-positive acute lymphoblastic leukemia Blood 1992 79: 3067–3070

    CAS  PubMed  Google Scholar 

  7. Chao NJ, Blume KG, Forman SJ, Snyder DS . Long-term follow-up of allogeneic bone marrow recipients for Philadelphia chromosome-positive acute lymphoblastic leukemia Blood 1995 85: 3353–3354

    CAS  PubMed  Google Scholar 

  8. Stockschläder M, Hegewisch S, Krüger W et al. Outcome of 14 patients with Philadelphia chromosome-positive ALL receiving autologous or allogeneic bone marrow transplantation Bone Marrow Transplant 1995 15: S71 (Abstr. 306)

    Google Scholar 

  9. Radich J, Gehly G, Lee A et al. Detection of bcr-abl transcripts in Philadelphia chromosome-positive acute lymphoblastic leukemia after marrow transplantation Blood 1997 89: 2602–2609

    CAS  PubMed  Google Scholar 

  10. Sierra J, Radich J, Hansen JA et al. Marrow transplants from unrelated donors for treatment of Philadelphia chromosome-positive acute lymphoblastic leukemia Blood 1997 90: 1410–1414

    CAS  PubMed  Google Scholar 

  11. Martin H, Atta J, Fauth F et al. Autologous BMT/PBSCT in patients with BCR-ABL-positive acute lymphoblastic leukemia Bone Marrow Transplant 1998 21: S2 (Abstr. 5)

    Google Scholar 

  12. Brenner MK, Rill DR, Moen RC et al. Gene-marking to trace origin of relapse after autologous bone marrow transplantation Lancet 1993 341: 85–86

    Article  CAS  PubMed  Google Scholar 

  13. Martin H, Atta J, Zumpe P et al. Purging of peripheral blood stem cells yields BCR-ABL-negative autografts in patients with BCR-ABL-positive acute lymphoblastic leukemia Exp Hematol 1995 23: 1612–1618

    CAS  PubMed  Google Scholar 

  14. Atta J, Martin H, Bruecher J et al. Residual disease and immunomagnetic bead purging in patients with BCR-ABL-positive acute lymphoblastic leukemia Bone Marrow Transplant 1996 18: 541–548

    CAS  PubMed  Google Scholar 

  15. Maurer J, Bartram CR, Ludwig WD et al . Frequency of BCR/ABL m-RNA in adult lymphoblastic leukemia (ALL) as determined in a prospective study Ann Hematol 1993 67: (Suppl.) A80 (Abstr. 312)

    Google Scholar 

  16. Yokota S, Hansen-Hagge TE, Ludwig WD et al. Use of polymerase chain reactions to monitor minimal residual disease in acute lymphoblastic leukemia patients Blood 1991 77: 331–339

    CAS  PubMed  Google Scholar 

  17. Gobrecht O, Gobel U, Graubner U et al. Effect of dose intensity and therapy-induced leukocytopenia in intensive therapy on the prognosis of acute lymphatic leukemia in childhood. Results in 213 patients of the COALL-85 study Klin Padiatr 1992 204: 230–235

    Article  CAS  PubMed  Google Scholar 

  18. Mitterbauer G, Fodinger M, Scherrer R et al. PCR-monitoring of minimal residual leukaemia after conventional chemotherapy and bone marrow transplantation in BCR-ABL-positive acute lymphoblastic leukaemia (published erratum appears in Br J Haematol 1995; 90: 492) Br J Haematol 1995 89: 937–941

    Article  CAS  PubMed  Google Scholar 

  19. Funderud S, Erikstein B, Asheim HC et al. Functional properties of CD19+ B lymphocytes positively selected from buffy coats by immunomagnetic separation Eur J Immunol 1990 20: 201–206

    Article  CAS  PubMed  Google Scholar 

  20. Campana DE, Janossy G . Leukemia diagnosis and testing of complement fixing antibodies for bone marrow purging in ALL Blood 1986 68: 1264–1271

    CAS  PubMed  Google Scholar 

  21. Kvalheim G, Sorensen O, Fodstad O et al. Immunomagnetic removal of B-lymphoma cells from human bone marrow: a procedure for clinical use Bone Marrow Transplant 1988 3: 31–41

    CAS  PubMed  Google Scholar 

  22. Wang MY, Fodstad O, Ludwig WD et al. An effective, direct immunomagnetic procedure for purging acute lymphoblastic leukemia cells from human bone marrow Pathol Oncol Res 1995 1: 32–37

    Article  PubMed  Google Scholar 

  23. Pegoraro L, Matera L, Ritz J et al. Establishment of a Ph1-positive human cell line (BV173) J Natl Cancer Inst 1983 70: 447–451

    CAS  PubMed  Google Scholar 

  24. Dhut S, Gibbons B, Chaplin T, Young BD . Establishment of a lymphoblastoid cell line, SD-1, expressing the p190 bcr-abl chimaeric protein Leukemia 1991 5: 49–55

    CAS  PubMed  Google Scholar 

  25. Chomczynski P, Sacchi N . Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction Anal Biochem 1987 162: 156–159

    Article  CAS  PubMed  Google Scholar 

  26. Martin H, Atta J, Bruecher J et al. In patients with BCR-ABL-positive ALL in CR peripheral blood contains less residual disease than bone marrow: implications for autologous BMT Ann Hematol 1994 68: 85–87

    Article  CAS  PubMed  Google Scholar 

  27. Tsuchiya H, Adachi N, Asou N et al. Responses to granulocyte colony-stimulating factor (G-CSF) and granulocyte–macrophage CSF in Ph1-positive acute lymphoblastic leukemia with myeloid surface markers (letter) (see comments) Blood 1991 77: 411–413

    CAS  PubMed  Google Scholar 

  28. Komada Y, Zhou YW, Zhang SL et al. Role in growth regulation of cytokines and cytokine receptors in acute lymphoblastic leukaemia expressing myeloid markers Br J Haematol 1993 84: 408–415

    Article  CAS  PubMed  Google Scholar 

  29. van Rhee F, Marks DI, Lin F et al. Quantification of residual disease in Philadelphia-positive acute lymphoblastic leukemia: comparison of blood and bone marrow Leukemia 1995 9: 329–335

    CAS  PubMed  Google Scholar 

  30. Miyamura K, Tanimoto M, Morishima Y et al. Detection of Philadelphia chromosome positive acute lymphoblastic leukemia by polymerase chain reaction: possible eradication of minimal residual disease by marrow transplantation Blood 1992 79: 1366–1370

    CAS  PubMed  Google Scholar 

  31. Brennan C, Weisdorf D, Kersey J et al. Bone marrow transplantation for Philadelphia chromosome-positive acute lymphoblastic leukemia Proc ASCO 1991 10: 222

    Google Scholar 

  32. Fabrega S, Laporte JP, Giarratana MC et al. Polymerase chain reaction: a method for monitoring tumor cell purge by long-term culture in BCR/ABL positive acute lymphoblastic leukemia Bone Marrow Transplant 1993 11: 169–173

    CAS  PubMed  Google Scholar 

  33. Grande M, Barbu V, van den Akker J et al. Autologous bone marrow transplantation in ALL: relapse linked to infusion of tumor cells with the back-up marrow Bone Marrow Transplant 1994 14: 477–480

    CAS  PubMed  Google Scholar 

  34. Gehly GB, Bryant EM, Lee AM et al. Chimeric BCR-abl messenger RNA as a marker for minimal residual disease in patients transplanted for Philadelphia chromosome-positive acute lymphoblastic leukemia Blood 1991 78: 458–465

    CAS  PubMed  Google Scholar 

  35. Annino L, Ferrari A, Cedrone M et al. Adult Philadelphia-chromosome-positive acute lymphoblastic leukemia: experience of treatments during a ten-year period Leukemia 1994 8: 664–667

    CAS  PubMed  Google Scholar 

  36. Preti HA, O'Brien S, Giralt S et al. Philadelphia-chromosome-positive adult acute lymphocytic leukemia: characteristics, treatment results, and prognosis in 41 patients Am J Med 1994 97: 60–65

    Article  CAS  PubMed  Google Scholar 

  37. Carella AM, Frassoni F, Pollicardo N et al. Philadelphia-chromosome-negative peripheral blood stem cells can be mobilized in the early phase of recovery after a myelosuppressive chemotherapy in Philadelphia-chromosome-positive acute lymphoblastic leukaemia Br J Haematol 1995 89: 535–538

    Article  CAS  PubMed  Google Scholar 

  38. Dunlop LC, Powles R, Singhal S et al. Bone marrow transplantation for Philadelphia chromosome-positive acute lymphoblastic leukemia Bone Marrow Transplant 1996 17: 365–369

    CAS  PubMed  Google Scholar 

  39. Wang MY, Fodstad O, Funderud S et al. A direct and effective immunomagnetic method for bone marrow purging in non-T ALL malignancies. The use of an anti-HLA-DR antibody gives sustained engraftment Prog Clin Biol Res 1994 389: 119–124

    CAS  PubMed  Google Scholar 

  40. Vormoor J, Baersch G, Baumann M et al. Flow cytometric identification of candidate normal stem cell populations in CD45-negative B-cell precursor acute lymphoblastic leukaemia Br J Haematol 1998 100: 501–508

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Deutsche Forschungsgemein- schaft (Ho 684/2-1). We appreciate the contribution of the hematologists participating in the German multicenter adult ALL studies.

Author information

Authors and Affiliations

  1. Department of Hematology and Oncology, Johann Wolfgang Goethe-University, Frankfurt/Main, Germany

    J Atta, F Fauth, C Weber, G Lippok, D Hoelzer & H Martin

  2. Institute of Transfusion Medicine, Johann Wolfgang Goethe-University, Frankfurt/Main, Germany

    M Keyser & E Petershofen

Authors
  1. J Atta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F Fauth
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M Keyser
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. E Petershofen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C Weber
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. G Lippok
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. D Hoelzer
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. H Martin
    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

Atta, J., Fauth, F., Keyser, M. et al. Purging in BCR-ABL-positive acute lymphoblastic leukemia using immunomagnetic beads: comparison of residual leukemia and purging efficiency in bone marrow vs peripheral blood stem cells by semiquantitative polymerase chain reaction. Bone Marrow Transplant 25, 97–104 (2000). https://doi.org/10.1038/sj.bmt.1702096

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.bmt.1702096

Keywords

Search

Advanced search

Quick links