1. Materials and methods:
Immunophenotypic analysis of multiple myeloma patients at the diagnosis until autologous stem cell transplantation. F. Akhundova, L. Mendeleeva, S.Lugovskaya, E. Naumova, A. Shcherbakova, E. Parovichnikova,V. Savchenko. Research Center for Hematology (Moscow, RU), Russian Medical Academy of Postgraduate Educaton (Moscow, RU).
Introduction
Results:
Immunophenotyping (IPT), using multiparametric flow cytometry (MFC) nowadays is demanded and necessary method in oncohematology. IFT by MFC is a very sensitive and powerful approach, corresponding to the detection level of neoplastic cells 1:104, permitting us to consider this method more sensitive than others for evaluating minimal residual disease (MRD).
The objective of this study was definition of dynamics of quantitative expression of aberrant antigens, such as CD 19, CD 117, CD 56 on myeloma plasma cells (M-PCs) CD 138+/CD38+ at the diagnosis until undergoing autologous stem cell transplantation (ASCT), using MFC.
IPT were performed 24 patients with newly diagnosed MM. Autologous stem cells were prepared for 8 of them; 5 of them were undergoing 1st ASCT, and 3 of these 5 them were undergoing 2nd ASCT. Among remaining 16 initial patients, 6 relapsed and 10 refused ASCT.
Antigen title
At the
diagnosis
Before SCM
b/m, %
During SCM CPSC, %
Prior 1st
ASCT, %
Prior 2nd
138/38
19.32±4.05
( )
0.57±0.17 ( )
0.27±0.07 ( )
0.92±0.37 ( )
0.59±0.08 ( )
138/19
26.74±7.84
( )
32.48±14.44( )
1.89
42.89± ( )
11.72± ( )
38/19
26.29±7.84
32.76± ( )
3.58±1.69 ( )
40.45± ( )
11.72± ( )
138/117
42.84±8.71
( )
30.88± ( )
34.67± ( )
15.23± ( )
38/117
42.08±8.33
36.31± ( )
15.27± ( )
138/56
80.88±7.3
( )
74.15± ( )
100.0
( )
51.82± ( )
52.97± ( )
38/56
81.51±6.97
( )
73.98± ( )
51.35± ( )
52.64± ( )
Materials and methods:
We have conducted prospective analysis of 24 newly diagnosed patients with multiple myeloma (MM) between November 2010 and June Among them were14 males and 10 females, age (50.6±2.45). Diagnosis was determined according to International Myeloma Working Group criteria (IMWG 2008). IPT of bone marrow cells was performed at the diagnosis stage; before stem cell mobilization (SCM) i.e. after completion of induction chemotherapy (ICT),( included 4-6 cycles of treatment regimens: VD, VCD, PAD); during SCM stage ( we used concentrate of peripheral blood stem cells (CPSC), harvested for ASCT) and prior to 1st and 2nd ASCT. SCM included cyclophosphamide 4 g/m2 plus G-CSF 5µg/kg/day. Conditioning prior to ASCT included melphalan 200 mg/m2.
Quantitation of neoplastic cells was performed by using Beckman Coulter FC-500 flow cytometer. IPT studies were performed on erythrocyte-lysed BM aspirate and samples of CPSC, using 4-colour panel: CD 138 FITC/38 PE/45 PC7/19 PerCP; CD 138 FITC/38 PE/CD45 PC7/СD 117 PerCP (c-kit)-Cy5.5; CD 138 FITC/38 PerCP-Cy 5.5/CD 45 PC7/56 PE (monoclonal antibodies BD Biosciences). M-PCs were identified according to CD 138+/CD38+ coexpression. IPT of newly diagnosed patients was performed based on events, versus patients during high-dose chemotherapy based on events. All the IPT results were compared to immunofixation and protein electrophoresis of serum and urine and with morphology assessment of bone marrow aspirates.
Statistical interpretation of the results, represented in the table, was analysed using MS EXCEL program, data presented as M±m.
Image 1. At the diagnosis ( )
Image 2. Before ASCT ( )
Conclusion:
Detection of M-PCs at the diagnosis and monitoring of MRD during and after the high-dose treatment allows us to evaluate the effect of performed treatment, prevent the relapse of the disease; as well as dynamic monitoring for aberrant antigen expression allows us to determine the prognostic value of the disease. According to the results of immunophenotyping we can plan the future treatment or modify therapy standards.
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Utility of nine-color, 11-Parameter flow cytometry for detection of plasma cells neoplasms: A comparison with bone marrow morphologic findings and concurent
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