Calorimetric study as a potential test for choosing treatment of B-cell chronic lymphocytic leukemia

Toward personalized therapy for chronic lymphocytic leukemia

“…1C). The patient's overall reactivity manifested by the changes of parameters related to apoptosis as well as the observation of strong decreases in thermal transition at 95 ± 3°C are consistent with our previously reported data, 18 and consistent with our results reflecting a high-basal Noxa expression in PBMC cells of patient PZ, who reached CR after RCC administration. However, we have observed a low expression of Noxa before drug exposure of patient RJ's cells and its lack after cell exposure to RCM.…”

“…The limited data concerning apoptotic gene expression changes in CLL cells, mainly obtained from in vivo/ex vivo treatment. [16][17][18] In vitroreported studies were conducted on CLL cells exposed to purine analogs and directed mainly to p53-related genes and drug resistance. 16,17 The aim of the present study was to evaluate the ex vivo potential of apoptosis induction in primary tumoral peripheral blood mononuclear cells (PBMCs) by RCM, i.e., rituximab + cladribine + mafosfamide (ex vivo active form of cyclophosphamide) using Vybrant Apoptosis Assay #4, differential scanning calorimetry and western blot (Mcl-1, Bcl-2, procaspase-3, Noxa).…”

“…or with combination of cladribine (C), mafosfamide (M) and rituximab (R) for 48 h, at concentrations described previously. 18,32 Determination of living cell numbers was performed by flow cytometry using Vybrant Apoptosis Assay #4 (Molecular Probes, Inc.). Preparation of whole-cell lysates and nuclear fraction was conducted as previously reported.…”

“…Preparation of whole-cell lysates and nuclear fraction was conducted as previously reported. 18 The control and drug-treated CLL cells were lysed. DSC analysis of the nuclear fraction isolated from leukemic PBMCs after incubation for 48 h with or without anticancer agents was made as described previously.…”

“…DSC analysis of the nuclear fraction isolated from leukemic PBMCs after incubation for 48 h with or without anticancer agents was made as described previously. 18 Nuclear were performed as previously described. 30 Sequence data were made using the international ImMunoGeneTics (IMGT) information system database and tools: the sequence with a nucleotide identity of 100% as compared with the germline was considered unmutated, while the sequence with an identity of 97.57% as mutated.…”