Stimulation of B-cell lymphoproliferations with CpG-oligonucleotide DSP30 plus IL-2 is more effective than with TPA to detect clonal abnormalities

Genetics of Chronic Lymphocytic Leukemia: Practical Aspects and Prognostic Significance

“…Other groups confirmed an improvement of the aberration detection rate in CpG/IL-2 (i.e. an increase of 9-13% of cases with aberrations) compared with TPA stimulated cultures (Put et al, 2009a;Struski et al, 2009). Moreover, the detection of translocations and del(13q) in particular, has been found to be superior after CpG/IL-2 stimulation compared with TPA (Put et al, 2009a).…”

“…The addition of the cytokine IL-2 to TPA stimulated CLL cell cultures was reported to directly stimulate CLL proliferation, even in absence of T-lymphocytes (Touw and Lowenberg, 1985). Although the latter findings provide evidence for the addition of IL-2 to TPA cultures, it is not mandatory for successful CCA (Put et al, 2009a;Struski et al, 2009). …”

“…The influence of CpG/IL-2 on quality of banding and metaphase generation is not clear (Put et al, 2009a;Struski et al, 2009).…”

“…Recently, improved culture methods have been introduced, i.e. CD40 ligand (CD40L)-induced cell cycle stimulation, and the immunostimulatory CpG oligonucleotide (DSP30) (Dicker et al, 2006;Haferlach et al, 2007;Mayr et al, 2006;Put et al, 2009a;Struski et al, 2009). …”

B-chronic lymphocytic leukemia chemoresistance involves innate and acquired leukemic side population cells

“…For proliferation assay, cells were labeled with 1 mM of carboxyfluorescein succinimidyl ester (Invitrogen, Cergy Pontoise, France) at 37 1C during 10 min and were subsequently treated or not with 250 nM of DSP30 (CpG-ODN) (Sequence: 5 0 -TCGTCGCTGTCTCCGCTTCTTCTTGCC-3 0 ) (TIB MOLBIOL, Berlin, Germany) in combination with 50 ng/ml of recombinant glycosylated interleukin (IL)-2 (kind gift from SanofiAventis, Toulouse, France) in RPMI containing 10% fetal calf serum during 7 days. 23,24 Flow cytometry…”