Journal: Pharmaceutics

Article Title: Enhanced Antitumor Efficacy of Cytarabine and Idarubicin in Acute Myeloid Leukemia Using Liposomal Formulation: In Vitro and In Vivo Studies

doi: 10.3390/pharmaceutics16091220

Figure Lengend Snippet: IC 50 values of liposomes and combined free drugs in CCRF-CEM cells.

Article Snippet: The results showed that the IC50 of the free cytarabine-idarubicin combination on CCRF-CEM cells was 216.12 nM, while the inhibitory effect of liposomes on the cells was significantly weaker than that of free drugs with an IC50 value of 5826 nM ( , A,B), because the drugs in the liposomes must be released gradually rather than directly interacting with cells to exert their killing effect.

Techniques: Liposomes

Journal: International Journal of Molecular Sciences

Article Title: C-C Chemokine Receptor 7 Promotes T-Cell Acute Lymphoblastic Leukemia Invasion of the Central Nervous System via β2-Integrins

doi: 10.3390/ijms25179649

Figure Lengend Snippet: Characterization of CCL19, CCL21, and CCL19 8-83 in CCR7-CEM cells. Chemotaxis of CCRF-CEM cells was assessed using a Boyden Chamber assay. ( A ) CCL19 or ( B ) CCL21 at the indicated concentrations or 50 nM ( C ) CCL19 or ( D ) CCL21 was added to the lower wells. CCRF-CEM cells ( A , B ) alone or ( C , D ) with CCL19 8-83 at the indicated concentration in the upper wells. Cells were allowed to migrate for 3 h across fibronectin-coated, 5 µm pore polycarbonate membranes. The percentage of cells migrated [(number of cells loaded/number of cells in bottom well) × 100] is shown. CCRF-CEM cells were induced to internalize ( E ) CCR7/CCL19 or ( F ) CCR7/CCL21 for the indicated times ± CCL19 8-83 and CCR7 remaining on the cell surface was assayed by flow cytometry. Data points represent the mean ±SD for n = 3+ independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 as determined using one-way ANOVA ( A – D ) or two-way ANOVA ( E , F ).

Article Snippet: Calcium Mobilization assays CCRF-CEM cells were resuspended at 1.5 × 10 7 cells/mL in Hanks Balanced Salt Solution (HBSS)/Fluo-4 AM (Invitrogen, F14201) for 45 min with gentle inversion every 5 min. CCL19 8-83 was added for five minutes to the blocked cells, and the cells were stimulated with either CCL19 or CCL21.

Techniques: Chemotaxis Assay, Boyden Chamber Assay, Concentration Assay, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: C-C Chemokine Receptor 7 Promotes T-Cell Acute Lymphoblastic Leukemia Invasion of the Central Nervous System via β2-Integrins

doi: 10.3390/ijms25179649

Figure Lengend Snippet: CCL19 8-83 blocks CCL19-induced calcium mobilization. ( A , D , G ) CCRF-CEM cells were pre-loaded with Fluo-4 AM at 37 °C, rinsed, and pre-incubated with CCL19 8-83 for five minutes before being stimulated with CCL19, CCL21, or CCL19 8-83 . Calcium mobilization was normalized to the ionomycin control maximum; data are plotted as the percentage of ionomycin control. ( B , E , H ), the maximal peak normalized percentage of Ca 2+ mobilized and ( C , F , I ), time (seconds) to reach peak Ca 2+ mobilization. Data points represent the mean ± SD of n = 3 independent experiments. ** p < 0.01 (unpaired two-tailed Student’s t -test).

Article Snippet: Calcium Mobilization assays CCRF-CEM cells were resuspended at 1.5 × 10 7 cells/mL in Hanks Balanced Salt Solution (HBSS)/Fluo-4 AM (Invitrogen, F14201) for 45 min with gentle inversion every 5 min. CCL19 8-83 was added for five minutes to the blocked cells, and the cells were stimulated with either CCL19 or CCL21.

Techniques: Incubation, Control, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: C-C Chemokine Receptor 7 Promotes T-Cell Acute Lymphoblastic Leukemia Invasion of the Central Nervous System via β2-Integrins

doi: 10.3390/ijms25179649

Figure Lengend Snippet: CCL19 8-83 inhibits CCL19-induced chemotaxis of CCRF-CEM across an intact HBEC-5i monolayer. ( A ) Trans-Endothelial Electrical Resistance (TEER) was measured every two days over 10 days. Data points represent the mean ± SD; n = 9. ( B ) HBEC-5i were plated on glass coverslips and allowed to form monolayers over several days. Cells were permeabilized and stained with anti-ZO-1-Alexa Fluor 647 (direct conjugate) and nuclei were stained with Hoechst and imaged at 40X (upper panel) or digitally zoomed (lower panel) to observe tight junctions. ( C ). T-ALL cell trans-endothelial cell migration to CCL19 but not CCL21 was inhibited in the presence of CCL19 8-83. Cells were pre-incubated in the presence of CCL19 8-83 and allowed to migrate to CCL19 across an HBEC-5i monolayer for 16 h, and cells that migrated to the lower well were counted. Data is the mean ± SD of n = 3 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Scale bar = 10 µm.

Article Snippet: Calcium Mobilization assays CCRF-CEM cells were resuspended at 1.5 × 10 7 cells/mL in Hanks Balanced Salt Solution (HBSS)/Fluo-4 AM (Invitrogen, F14201) for 45 min with gentle inversion every 5 min. CCL19 8-83 was added for five minutes to the blocked cells, and the cells were stimulated with either CCL19 or CCL21.

Techniques: Chemotaxis Assay, Staining, Migration, Incubation

Journal: International Journal of Molecular Sciences

Article Title: C-C Chemokine Receptor 7 Promotes T-Cell Acute Lymphoblastic Leukemia Invasion of the Central Nervous System via β2-Integrins

doi: 10.3390/ijms25179649

Figure Lengend Snippet: CCL19 8-83 inhibits β2 integrin-mediated T-ALL migration across a cerebral microvascular endothelial cell monolayer. CCRF-CEM cells were pre-incubated with 10 µg/mL AIIB2 (anti-β1 integrin) ± CCL19 8-83 or 10 µg/mL CBR-LFA1/2 (anti-β2 integrin) for five minutes prior to loading in the top well. Cells were allowed to invade and transmigrate over 16 h, and migrated cells normalized to the number of cells that migrated to CCL19 alone. n = 3 independent experiments. n = 3 independent experiments. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (unpaired two-tailed Student’s t -test).

Article Snippet: Calcium Mobilization assays CCRF-CEM cells were resuspended at 1.5 × 10 7 cells/mL in Hanks Balanced Salt Solution (HBSS)/Fluo-4 AM (Invitrogen, F14201) for 45 min with gentle inversion every 5 min. CCL19 8-83 was added for five minutes to the blocked cells, and the cells were stimulated with either CCL19 or CCL21.

Techniques: Migration, Incubation, Two Tailed Test