Quantification of chimerism by real-time polymerase chain reaction.

Czech version

Authors: O. Horký; J. Mayer; D. Dvořáková
Authors‘ workplace: Centrum molekulární biologie a genové terapie, Interní hematoonkologická klinika, Fakultní nemocnice Brno
Published in: Transfuze Hematol. dnes,13, 2007, No. 2, p. 73-78.
Category: Comprehensive Reports, Original Papers, Case Reports

Overview

Real time polymerase chain reaction (RQ-PCR) represents one of the most sensitive methods for quantitative analysis of chimerism. This method is at least ten times more sensitive than commonly performed method of fragment analysis (FA) with fluorescence detection. Application of method that enables detection of microchimerism (mixed chimerism under 1%), can recognize relaps significantly sooner and in more cases than by using less sensitive methods. Performance of RQ-PCR within patients with sudden recurrence of autologous haematopoiesis showed microchimerism also in previous samples, formerly judged according to FA as complete donor chimerism (CDC). Furthermore, retrospective analysis of CDC samples from patient with positive residual disease repeatedly revealed microchimerism. Performance of more sensitive method can significantly contribute to early clinical intervention, nevertheless the predictable value of mixed chimerism and especially microchimerism must be further investigated.

Key words:
chimerism, microchimerism, real-time PCR, fragment analysis

Sources

1. Thomas ED. Bone marrow transplantation: a review. Semin Hematol 1999; 36: 95–103.

2. Antin JH, Childs R, Filipovich AH, et al. Establishment of complete and mixed donor chimerism after allogeneic lymphohematopoietic transplantation: recommendations from a workshop at the 2001 Tandem Meetings of the International Bone Marrow Transplant Registry and the American Society of Blood and Marrow Transplantation. Biol Blood Marrow Transplant 2001; 7: 473–485.

3. Thiede C, Bornhäuser M, Ehninger G. Strategies and clinical implications of chimerism diagnostics after allogeneic hematopoietic stem cell transplantation. Acta Haematol 2004; 112: 16–23.

4. Bader P, Niethammer D, Willasch A, Kreyenberg H, Klingebiel T. How and when should we monitor chimerism after allogeneic stem cell transplantation? Bone Marrow Transplant 2005; 35: 107–119.

5. Horký O, Dvořáková D, Mayer J. Kvantitativní analýza chimerismu po alogenní transplantaci hematopoetických buněk. Trans Hemat dnes 2004; 10: 70–75.

6. Lion T. Summary: Reports on quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection. Leukemia 2003; 17: 252–254.

7. Alizadeh M, Bernard M, Danic B, et al. Quantitative assessment of hematopoietic chimerism after bone marrow transplantation by real-time quantitative polymerase chain reaction. Blood 2002; 99: 4618–4625.

8. Elmaagacli AH, Hlinka M, Steckel NC, Beelen DW.Anew sensitive quantitative real-time PCR method for chimerism analyses can help to identify patients at high risk for leukemic relapse after allogeneic stem cell transplant earlier than STR-PCR chimerism analyses. Bone Marrow Transplant 2003; S123

9. Thiede C, Kellermann T, Schwerdtfeger R, et al. Real-time PCR for the SRY-gene allows sensitive and quantitative chimerism analysis after allogeneic blood stem cell transplantation: clinical results in 43 patients. Bone Marrow Transplant 2003; 31: S31.

10. Uzunel M, Sundin U, Mattsson J, Schaffer M, Hauzenberger D, Ringden O. Sensitive and quantitative chimerism analysis in ALL patients with real-time PCR. Bone Marrow Transplant 2004; 33: S101.

11. Baak U, Marinets O, Rieger O, et al. Real-time quantitative Y chromosome specific PCR vs. XY-FISH vs. fluorescent STRPCR for chimerism analysis after allogeneic sex-mismatched PBSCT. ASH Annnual Meeting Abstracts 2004; 104: 5047

12. Jiménez-Velasco A, Barrios M, Román-Gómez J, et al. Reliable quantification of hematopoietic chimerism after allogeneic transplantation for acute leukemia using amplification by realtime PCR of null aleles and insertion/deletion polymorphisms. Leukemia 2005; 19: 336–343.

13. Kroger N, Zagrivnaja M, Schwartz S, et al. Kinetics of plasmacell chimerism after allogeneic stem cell transplantation by highly sensitive real-time PCR based on sequence polymorphism and its value to quantify minimal residual disease in patients with multiple myeloma. Exp Hematol 2006; 34: 688–694.

14. Koldehoff M, Steckel NK, Hlinka M, Beelen DW, Elmaagacli AH. Quantitative analysis of chimerism after allogeneic stem cell transplantation by real time polymerase chain reaction with single nucleotide polymorphism, standard tandem repeats, and Y chromosome-specific sequences. Am J Hematol 2006; 81: 735–746.

15. Oliver DH, Thompson RE, Griffin CA, Eshleman JR. Use of single nucleotide polymorphisms (SNP) and real-time polymerase chain reaction for bone marrow engraftment analysis. J Mol Diagn 2000; 2: 202–208.

16. Sachidanandam R, Weissman D, Schmidt SC, et al. A map of human genome sequence variation containing 1.42 million single nucleotide polymorphisms. Nature 2001; 409: 928–933.

17. Fehse B, Chukhlovin A, Kuhlcke K, et al. Real-time quantitative Y chromosome-specific PCR (QYCS-PCR) for monitoring hematopoietic chimerism after sex-mismatched allogeneic stem cell transplantation. J Hematother Stem Cell Res 2001; 10: 419–425.

18. Wilhelm J, Reuter H, Tews B, Pingoud A, Hahn M. Detection and quantification of insertion/deletion variations by allele-specific real-time PCR: application for genotyping and chimerism analysis. Biol Chem 2002; 383: 1423–1433.

19. Naparstek E, Vainas O, Eshel R. Real-time PCR SNP assay for monitoring chimerism after allogeneic stem cell transplantation. Blood 2003; 102: Abstract 5460.

20. Maas F, Schaap N, Kolen S, et al. Quantification of donor and recipient hemopoietic cells by real-time PCR of single nucleotide polymorphisms. Leukemia 2003; 17: 621–629.

21. Lucas KG, Sun Q, Chorney K, Erickson T, Nelson JL. Microchimerism detection by HLA-specific real-time polymerase chain reaction analysis in recipients of allogeneic Epstein Barr virus specific cytotoxic T lymphocytes. ASH Annual Meeting Abstracts 2004; 104: 2648.

22. Rabascio C, Raia V, Muratori E, et al. Quantitative PCR for chimerism kinetics in non-myeloablative allo-transplants. Bone Marrow Transplant 2005; R1181.

23. Buno I, Barrios M, Navarro G, et al. A methodological algorithm for chimerism quantification after stem cell transplantation. Bone Marrow Transplant 2005; P598.

24. Jiménez-Velasco A, Barrios M, Roman-Gomez J, et al. Differential prognostic significance of bone marrow and peripheral blood mixed chimerism determined by real-time quantitative PCR after SCT in patients with acute leukemia. Bone Marrow Transplant 2005; P599.

25. Harries LW, Wickham CL, Evans JC, Rule SA, Joyner MV, Ellard S. Analysis of haematopoietic chimerism by quantitative real-time polymerase chain reaction. Bone Marrow Transplant 2005; 35: 283–290.

26. Masmas TN, Madsen HO, Petersen SL, et al. Evaluation and automation of hematopoietic chimerism analysis based on realtime quantitative polymerase chain reaction. Biol Blood Marrow Transplant 2005; 11: 558–566.

27. Masmas TN, Petersen SL, Madsen HO, et al. Kinetics of early donor chimerism after nonmyeloablative haematopoietic stem cell transplantation monitored with high-resolution real time quantitative PCR. ASH Annual Meeting Abstracts 2005; 106: 3669.

28. Halaburda K, Guz K, Orzinska A, et al. Real-time PCR for early chimerism evaluation in peripheral blood mononuclear cells of allogeneic stem cell recipients: potential value for prediction of acute graft-versus-host disease. Bone Marrow Transplant 2006; 37: S95.

29. Picardi A, Spagnoli A, Cordone I, et al. Reliable quantification of haematopoietic chimerism after allogeneic stem cell transplantation by real-time quantitative PCR analysis. Bone Marrow Transplant 2006; 37: S322.

30. Eshel R, Vainas O, Shpringer M, Naparstek E. Highly sensitive patient-specific real-time PCR SNP assay for chimerism monitoring after allogeneic stem cell transplantation. Lab Hematol 2006; 12: 39–46.

31. Bai L, Deng YM, Dodds AJ, Milliken S, Moore J, Ma DD. A SYBR green-based real-time PCR method for detection of haemopoietic chimerism in allogeneic haemopoietic stem cell transplant recipients. Eur J Haematol 2006; 77: 425–431.

32. Chiu RW, Murphy MF, Fidler C, Zee BC, Wainscoat JS, Lo YM. Determination of RhD zygosity: comparison of a double amplification refractory mutation system approach and a multiplex real-time quantitative PCR approach. Clin Chem 2001; 47: 667–672.

33. Formánková R, Sedláček P, Kršková L, Říhová H, Šrámková L, Starý J. Chimerism-directed adoptive immunotherapy in prevention and treatment of post-transplant relapse of leukemia in childhood. Haematologica 2003; 88: 117–118.

34. Formánková R, Honzátková L, Sieglová Z, Starý J, Sedláček P, Brdička R. Detailed monitoring of hematopoietic chimerism in a child treated by adoptive immunotherapy for high risk of relapse after BMT for acute myeloid leukemia. Bone Marrow Transplant 2000; 25: 453–456.

35. Formánková R, Honzátková L, Moravcová J, Sieglová Z, Dvořáková R, Nádvorníková S., Vítek A, Lukášová M, Starý J, Brdička R. Prediction and reversion of post-transplant relapse in patients with chronic myeloid leukemia using mixed chimerism and residual disease detection and adoptive immunotherapy. Leuk Res 2000; 24: 339–347.

36. Baron F, Sandmaier BM. Chimerism and outcomes after allogeneic hematopoietic cell transplantation following nonmyeloablative conditioning. Leukemia 2006; 20: 1690–1700.

37. Khan F, Agarwal A, Agarwal S. Significance of chimerism in hematopoietic stem cell transplantation: new variations on an old theme. Bone Marrow Transplant 2004; 34: 1–12.

38. Serrano J, Roman J, Sanchez J, et al. Molecular analysis of lineage-specific chimerism and minimal residual disease by RTPCR of p210(BCR-ABL) and p190(BCR-ABL) after allogeneic bone marrow transplantation for chronic myeloid leukemia: increasing mixed myeloid chimerism and p190(BCR-ABL) detection precede cytogenetic relapse. Blood 2000; 95: 2659–2665.

39. Roman J, Alvarez MA, Torres A. Molecular basis for therapeutic decisions in chronic myeloid leukemia patients after allogeneic bone marrow transplantation. Haematologica 2000; 85: 1072–1082.

40. Horký O, Mayer J, Borský M, Pospíšilová J, Krejčí M, Dvořáková D. Dynamics of lineage-specific chimerism in patients after non-myeloablative hematopoietic stem cell transplantation. Haematologica 2006; 91(S1): 304.

41. Elmaagacli AH. Real-time PCR for monitoring minimal residual disease and chimerism in patients after allogeneic transplantation. Int J Hematol 2002; Suppl. 2: 204–205.

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Article was published in

Transfusion and Haematology Today

2007 Issue 2