SULM – Schweizerische Union für Labormedizin | Union Suisse de Médecine de Laboratoire | Swiss Union of Laboratory Medicine

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T.A. Ermakova1 , A.G. Turkina1 , N.V. Tsvetaeva1 , E.U. Chelusheva1 , L.S. Dyachenko1 , N.N. Tsyba1 , S.V. Kolodey1 , N.D. Khoroshko1

1National Research Center for Hematology,Russia

Objectives: the goal was to distinguish between mono and polyclonal hematopoiesis in chronic myeloid leukemia (CML) patients and in patients with other hematological diseases (HD-s) for assessing a trustfull therapy and prognosis of the disease.
Patients and Methods: the study included a total of 18 controls and 42 patients comprising 4 Ph(+) and 10 Ph(-) CML patients (Philadelphia-chromosome positive or negative).Ten CML Ph(+) patients previously treated with Imatinib or Interferon were observed in ongoing clinical and hematological remission. Five patients taking Imatinib demonstrated a response in 6 - 12 months and 2 patients in 3 months.We applied 3 methods to prove the restoration of Ph(-) hematopoiesis in leukocytes (leu), bone marrow (b/m): 1. HUMARA-assay to evaluate clonality;2.Real-time PCR (qPCR) to measure levels of bcr/abl gene; 3.Cytogenetic method to reveal Ph(+) cells.
For comparison we investigated patients with other HD-s including refractory anaemia, aplastic anaemia, partial red cell aplasia, lymphoma.To specify mono/polyclonal nature of hematopoiesis in leu we used the HUMARA-assay, evaluating the lesion ratios (l.r.) of methylated patterns of polymorphic sites of HUMARA gene located on X chromosome.
Results: the treatment of 10 CML patients with follow-up of 3-7 years led to a response of b/m based on restored hystology, cytogenetic data with 0.0-5.0% Ph(+) cells and complete molecular remission tested by qPCR. HUMARA-assay revealed polyclonal hematopoiesis with l.r.fluctuating between 0.59-1.33,normal range.Four Ph(+)CML patients showed monoclonality exceeding l.r. = 2.7. In other HD-s mono and polyclonality were observed.
Conclusion: restoration of Ph(-) hematopoiesis in CML female patients treated with Imatinib or Interferon can be detected by HUMARA-assay, equally well with qPCR and cytogenetic analysis and taken together give full picture of Ph(-) CML. Estimation of mono/polyclonal hematopoiesis is a preliminary prognostic tool in HD-s.

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