A comparison of flow cytometry detection of minimal residual disease and chimerism kinetics in chronic lymphocytic leukemia patients after allogeneic hematopoietic stem cell transplantation

Adriana Plesa, Xavier Thomas, Quoc Hung Le, Anne-Sophie Michallet, Valérie Dubois, Charles Dumontet, Mauricette Michallet

.

DOI: 10.4236/jbise.2011.43024                

Abstract

Determination of minimal residual disease (MRD) remains crucial for the follow-up after therapy in chronic lymphocytic leukemia (CLL) patients. Chimerism was assessed by short tandem repeat (STR)-PCR and single nucleotide polymorphisms (SNP)-PCR, and MRD by a multicolor flow cytometric approach in 12 consecutive patients with CLL after they received allogeneic stem cell transplantation (SCT). Overall, 11 patients achieved MRD flow negativity [10 had full donor chimerism (FDC) and one had mixed chimerism (MC)]. Only one patient remained with MRD flow positivity and displayed MC. Fifty-six samples were concomitantly studied by both chimerism and MRD flow. A significant correlation was observed between MRD flow data and chimerism in both PB and BM by using a mixed effect linear regression (p < 0.001). Flow cytometry approach of MRD can be easily combined with chimerism during the follow-up post-allogeneic SCT. Both techniques appeared complementary for guiding post-transplant immunomodulation.

Keywords

Chronic lymphocytic leukemia, allogeneic stem cell transplantation, minimal residual disease, chimerism

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Dighiero, G. and Hamblin, T.J. (2008) Chronic lymphocytic leukaemia. Lancet, 371, 1017-1029. doi:10.1016/S0140-6736(08)60456-0
[2]	Moreton, P., Kennedy, B., Lucas, G., Leach, M., Rassam, A.M.B., Haynes, A., et al. (2005) Eradication of minimal residual disease in B-cell chronic lymphocytic leukaemia after Alemtuzumab therapy is associated with prolonged survival. Journal of Clinical Oncology, 23, 2971-2979. doi:10.1200/JCO.2005.04.021
[3]	Maloum, K., Sutton, L., Baudet, S., Laurent, C., Bonnemye, P., Magnac, C., et al. (2002) Novel flow- cytometric analysis based on B CD5+ subpopulations for the evaluation of minimal residual disease in chronic lymphocytic leukaemia. British Journal of Haematology, 119, 970-975.
[4]	Dreger, P., Ritgen, M., Bottcher, S., Schmitz, N. and Kneba, M. (2005) The prognostic impact of minimal residual disease assessment after stem cell transplantation for chronic lymphocytic leukaemia: is achievement of molecular remission worthwhile? Leukemia, 19, 1135-1138. doi:10.1038/sj.leu.2403800
[5]	B?ttcher, S., Ritgen, M., Pott, C., Bruggemann, M., Raff, T., Stilgenbauer, S., et al. (2004) Comparative analysis of minimal residual disease detection using four-color flow cytometry, consensus IgH-PCR, and quantitative IgH PCR in CLL after allogeneic and autologous stem cell transplantation. Leukemia, 18, 1637-1645. doi:10.1038/sj.leu.2403478
[6]	Ritgen, M., B?ttcher, S., Stilgenbauer, S., Bunjes, D., Schubert, J., Cohen, S., et al. (2008) Quantitative MRD monitoring identifies distinct GVL response patterns after allogeneic stem cell transplantation for chronic lymphocytic leukemia: results from the GCLLSG CLL3X trial. Leukemia, 22, 1377-1386. doi:10.1038/leu.2008.96
[7]	Rawstron, A.C., Villamor, N., Ritgen, M., Bottcher, S., Ghia, P., Zehnder, J.L., et al. (2007) International standardized approach for flow cytometric residual disease monitoring in chronic lymphocytic leukemia. Leukemia, 21, 956-964.
[8]	Cheson, B.D., Bennett, J.M., Grever, M., Kay, N., Keating, M.J., O’Brien, S., et al. (1996) National Cancer Institured-sponsored Working Group guidelines for chronic lymphocytic leukemia: Revised guidelines for diagnosis and treatment. Blood, 87, 4990-4997.
[9]	Moreno, C., Villamor, N., Colomer, D., Esteve, J., Giné, E., Muntanola, A., et al. (2006) Clinical significance of minimal residual disease, as assessed by different tech- niques, after stem cell transplantation for chronic lym- phocytic leukemia. Blood, 107, 4563-4569. doi:10.1182/blood-2005-09-3634
[10]	Maloum, K., Charlotte, F., Divine, M., Cazin, B., Lesty, C., Merle-Béral, H., for the French Cooperative Group on CLL. (2006) A comparison of the sensitivity of flow cytometry and bone marrow biopsy in the detection of minimal residual disease in chronic lymphocytic leuke- mia. Haematologica, 91, 860-861.
[11]	Dreger, P., Corradini, P., Kimby, E., Michallet, M., Milligan, D., Schetelig, J., et al. (2007) Chronic Leukemia Working Party of the EBMT. Indications for allogeneic stem cell transplantation in chronic lymphocytic leukemia: the EBMT transplant consensus. Leukemia, 21, 12-17. doi:10.1038/sj.leu.2404441
[12]	Binet, J.L., Auquier, A., Dighiero, G., Chastang, C., Piguet, H., Goasguen, J, et al. (1981) A new prognostic classification of chronic lymphocytic leukemia derived from a multivariate survival analysis. Cancer, 48, 198- 206.doi:10.1002/1097-0142(19810701)48:1<198::AID-CNCR2820480131>3.0.CO;2-V
[13]	Thiede, C., Florek, M., Bornhauser, M., Ritter, M., Mohr, B., Brendel, C., et al. (1999) Rapid quantification of mixed chimerism using multiplex amplification of short tandem repeat markers and fluorescence detection. Bone Marrow Transplant, 23, 1055-1060. doi:10.1038/sj.bmt.1701779
[14]	Alizadeh, M., Bernard, M., Danic, B., Dauriac, C., Birebent, B., Lapart, C., et al. (2002) Quantitative assessment of hematopoietic chimerism after bone marrow transplanta- tion by real-time quantitative polymerase chain reaction. Blood, 99, 4618-4625. doi:10.1182/blood.V99.12.4618
[15]	Pinheiro, J.C., Bates, D.M. (2000) Mixed-effects models in S and S-PLUS. Springer, New-York.
[16]	Keating, M. J., O’Brien, S., Albitar, M., Lerner, S., Plunkett, W., Giles, F., et al. (2005) Early results of a chemoim- munotherapy regimen of fludarabine, cyclophosphamide, and rituximab as initial therapy for chronic lymphocytic leukemia. Journal of Clinical Oncology, 23, 4079-4088. doi:10.1200/JCO.2005.12.051
[17]	Keating, M.J., Flinn, I., Jain, V., Binet, J.L., Hillmen, P., Byrd, J., et al. (2002) Therapeutic role of alemtuzumab (Campath-1H) in patients who have failed fludarabine: results of a large international study. Blood, 99, 3554- 3561. doi:10.1182/blood.V99.10.3554
[18]	Michallet, M., Archimbaud, E., Rowlings, P.A., Deeg, H. J., Gahrton, G., Montserrat, E., et al. (1996) HLA-iden- tical sibling bone marrow transplants for chronic lym- phocytic leukemia. Ann Intern Med, 124, 311-315.
[19]	Rabinowe, S. N., Soiffier, R. J., Gribben, J., Daley, H., Freedman, A. S., Daley, J., et al. (1993) Autologous and allogeneic bone marrow transplantation for poor progno- sis patients with B-cell chronic leukemia. Blood, 4, 1366- 1376.
[20]	Brown, J.R., Kim, H.T., Li, S., Stephans, K., Fisher, D.C., Cutler, C., et al. (2006) Predictors of improved progression-free survival after nonmyeloablative allogeneic stem cell transplantation for advanced chronic lymphocytic leukemia. Biol Blood Marrow Transplant, 12, 1056-1064. doi:10.1016/j.bbmt.2006.06.004
[21]	Khouri, I.F., Lee, M.S., Saliba, R.M., Anderlini, P., Couriel, D., Hosing, C., et al. (2004) Nonablative allogeneic stem cell transplantation for chronic lymphocytic leukemia: Impact of rituximab on immunomodulation and survival. Experimental Hematology, 32, 28-35. doi:10.1016/j.exphem.2003.09.021
[22]	Provan, D., Bartlett-Pandite, L., Zwicky, C., Neuberg, D., Maddocks, A., Corradini, P., et al. (1996) Eradication of PCR detectable chronic lymphocytic cells is associated with improved outcome after bone marrow transplantation. Blood, 88, 2228-2235.
[23]	Uzunel, M., Mattsson, J., Brune, M., Johansson, J.E., Aschan, J. and Ringden, O. (2003) Kinetics of minimal residual disease and chimerism in patients with chronic myeloid leukemia after nonmyeloablative conditioning and allogeneic stem cell transplantation. Blood, 101, 469- 472. doi:10.1182/blood-2002-02-0571