Chromosome Banding Analysis of Peripheral Blood Lymphocytes Stimulated with IL‑2 and CpG Oligonucleotide DSP30 in Patients with Chronic Lymphocytic Leukemia


Authors: K. Štěpanovská 1;  G. Vaňková 1;  V. Némethová 1;  L. Tomášiková 1;  P. Šmuhařová 1;  E. Divíšková 1;  V. Vallová 2;  P. Kuglík 2;  K. Plevová 1;  A. Oltová 1;  M. Doubek 1,3;  Š. Pospíšilová 1,3;  J. Mayer 1,3
Authors‘ workplace: Interní hematologická a onkologická klinika, Centrum molekulární bio­logie a genové terapie, LF MU a FN Brno2 Oddělení lékařské genetiky, laboratoř molekulární cytogenetiky, LF MU a FN Brno3 CEITEC –  Středoevropský technologický institut, MU, Brno 1
Published in: Klin Onkol 2013; 26(4): 263-270
Category: Original Articles

Overview

Background:
Chromosomal aberrations play an important role as prognostic factors in chronic lymphocytic leukemia (CLL). These aberrations are mostly detected by fluorescent in situ hybridization (FISH), as chromosomal banding analysis has been scarce due to low proliferative activity of malignant B-lymphocytes in vitro. In 2006, a new method using stimulation with IL-2 and CpG oligonucleotide DSP30 for metaphase generation in CLL was published [1]. The objective of our study was to verify the efficacy of stimulation and to evaluate if the method is suitable for routine diagnostics.

Patients and Methods:
In total, peripheral blood samples of 369 CLL patients were analyzed in parallel by chromosomal banding analysis and by FISH probes for 13q14, 11q22–23, CEP12 and 17p13.

Results:
Out of 369 patients, 307 (83%) were successfully stimulated for metaphase generation. Chromosomal aberrations were detected in 243 (79%) out of 307 patients evaluated by chromosomal banding analysis. Other aberrations that are not included into standard FISH panel were detected in patients’ karyotypes, e.g. del(6q), del(14q), t(14;18)(q32;q21), t(11;14)(q13;q32) and t(18;22)(q21;q11). One hundred and three (42%) patients showed complex aberrant karyotype not detected by FISH analysis.

Conclusion:
Stimulation with IL-2 and oligonucleotide DSP30 is an efficient method how to induce proliferation of malignant B-lymphocytes and allows detection of a substantial number of chromosomal aberrations in addition to those detected by standard FISH panel. Using this method in routine diagnostics is helpful particularly in identification of patients with complex aberrant karyotype.

Key words:
cytogenetics – chronic lymphocytic leukemia – interleukin-2 – CpG-ODN DSP30 – fluorescent in situ hybridization – cultivation – chromosome aberration


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