Journal: International Journal of Molecular Sciences

Article Title: Persistence of Chronic Lymphocytic Leukemia Stem-like Populations under Simultaneous In Vitro Treatment with Curcumin, Fludarabine, and Ibrutinib: Implications for Therapy Resistance

doi: 10.3390/ijms25041994

Figure Lengend Snippet: Clinical data of studied CLL patients.

Article Snippet: Immunophenotyping characterization was performed using the following monoclonal antibodies: CD19-AF700, CD19-APC, CD19-PE, CD19-PE-Cy7 (clone LT19) (Sysmex, Kobe, Japan); CD5-APC, CD5-PacB (clone L17F12) (Sysmex); CD38-PE (clone HIT2) (Sysmex); CD5-PE (clone UCHT2) (Beckton Dickinson, Franklin Lakes, NJ, USA).

Techniques: Diagnostic Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Persistence of Chronic Lymphocytic Leukemia Stem-like Populations under Simultaneous In Vitro Treatment with Curcumin, Fludarabine, and Ibrutinib: Implications for Therapy Resistance

doi: 10.3390/ijms25041994

Figure Lengend Snippet: Strategy of analysis of samples and primary cell cultures. Plots exemplifying data obtained at the flow cytometer. For every patient, 5 different analyses, corresponding to columns, were performed: time zero, directly analyzing peripheral blood; and control and treatments with curcumin, drug, and drug plus curcumin, analyzing primary cultures after incubation with corresponding treatments. For every analysis, three parameters corresponding to rows were obtained: cell membrane integrity (upper row), CD19 and CD5 expression (middle row), and ALP activity (lower row).

Article Snippet: Immunophenotyping characterization was performed using the following monoclonal antibodies: CD19-AF700, CD19-APC, CD19-PE, CD19-PE-Cy7 (clone LT19) (Sysmex, Kobe, Japan); CD5-APC, CD5-PacB (clone L17F12) (Sysmex); CD38-PE (clone HIT2) (Sysmex); CD5-PE (clone UCHT2) (Beckton Dickinson, Franklin Lakes, NJ, USA).

Techniques: Flow Cytometry, Control, Incubation, Membrane, Expressing, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Persistence of Chronic Lymphocytic Leukemia Stem-like Populations under Simultaneous In Vitro Treatment with Curcumin, Fludarabine, and Ibrutinib: Implications for Therapy Resistance

doi: 10.3390/ijms25041994

Figure Lengend Snippet: Curcumin synergistic effect combined with fludarabine and ibrutinib on CLL primary cell cultures. Primary cultures were directly established from CLL patients’ peripheral blood and incubated at 37 °C in 5% CO 2 with different treatments. Curcumin, fludarabine, and ibrutinib concentrations used were 5 µM, 10 µM, and 10 µM, respectively. Incubation time ranged from 2 to 6 days. ( A ) Curcumin showed no significant effect on the leukemic fraction of samples of the fludarabine group, nor alone, nor combined with the drug. Fludarabine showed a significant effect in reducing the leukemic fraction. ( B ) Fludarabine showed a strong significant effect in reducing leukemic cells with high ALP activity, and combination with curcumin showed a significant synergistic effect in increasing this reduction. Curcumin did not show a significant effect when used alone. ( C ) Curcumin showed no significant synergistic effect combined with ibrutinib on the leukemic fraction, which was strongly reduced by the drug. However, curcumin significantly reduced the leukemic fraction when used alone. ( D ) Curcumin and ibrutinib showed q significant effect in reducing leukemic cells with high ALP activity in the ibrutinib group. Curcumin produced a strong and significant synergistic effect when combined with ibrutinib. Analysis was performed using flow cytometry. Among cells with intact cell membranes (PI-negative), cells positive for both CD19 and CD5 were selectively identified (leukemic cells). From these, cells with high ALP activity (stem-like) were selected ( B , D ). Substantial interpatient variability is associated with clinical differences among patients. Results were statistically analyzed using non-parametric Student’s t -test (Wilcoxon test). Values obtained for curcumin or drugs are compared with values obtained for the control. Values obtained for drug combined with curcumin are compared with values obtained for drug without curcumin. * p < 0.05; ** p < 0.001.

Article Snippet: Immunophenotyping characterization was performed using the following monoclonal antibodies: CD19-AF700, CD19-APC, CD19-PE, CD19-PE-Cy7 (clone LT19) (Sysmex, Kobe, Japan); CD5-APC, CD5-PacB (clone L17F12) (Sysmex); CD38-PE (clone HIT2) (Sysmex); CD5-PE (clone UCHT2) (Beckton Dickinson, Franklin Lakes, NJ, USA).

Techniques: Incubation, Activity Assay, Produced, Flow Cytometry, Control