Journal: Leukemia & lymphoma

Article Title: CD40 ligand is necessary and sufficient to support primary diffuse large B-cell lymphoma cells in culture: a tool for in vitro preclinical studies with primary B-cell malignancies

doi: 10.3109/10428194.2011.654337

Figure Lengend Snippet: Expression of CD40 in primary dog B cell lymphomas. (A) A heat map of selected gene expression in dog B-cell and T-cell lymphomas. Expressions of genes, chosen for prototypical expression in B-cells and T-cells, were shown as a heat map where color represents fluorescence intensity of hybridized probes as indicated at the bottom and therefore the amount of gene transcript in any given sample. Generally, Red/Orange represents high expression, Yellow/Green represents moderate expression and Blue/Black represents little or no expression. (B) DLBCL samples were stained for CD22 and CD40 (using CD40L-CD8 fusion protein). A representative result using an isotype control antibody and streptavidin (SA control) is shown as a negative control (left panel). One representative sample of 3 different primary dog B-cell lymphomas is shown, including the mean fluorescence intensity (MFI) of the FITC channel.

Article Snippet: The human DLBCL cell line SU-DHL4 was obtained from Dr. Lee Honigberg (Pharmacyclics Inc., Sunnyvale, CA).

Techniques: Expressing, Gene Expression, Fluorescence, Staining, Control, Negative Control

Journal: Leukemia & lymphoma

Article Title: CD40 ligand is necessary and sufficient to support primary diffuse large B-cell lymphoma cells in culture: a tool for in vitro preclinical studies with primary B-cell malignancies

doi: 10.3109/10428194.2011.654337

Figure Lengend Snippet: In vitro culture of primary dog DLBCL cells with KtCD40L. (A) Photomicrographs of DLBCL cells alone (left), DLBCL cells cultured with KtCD40L (middle), and KtCD40L alone (right), each shown after seven days in culture. Bars = 100 μm (B) Flow analysis of DLBCL cells before and after culture with KtCD40L for 7 days. Estimates of cell size and complexity were obtained from flow cytometric light scatter properties (upper panels). Viability was determined by 7-AAD exclusion (lower panels). FSC; forward scatter, SSC; side scatter (C) Growth curves of primary dog DLBCL cells (n = 4) cultured with KtCD40L. DLBCL cell numbers were determined at each time point using hemacytometer with dead cell exclusion by trypan blue staining.

Article Snippet: The human DLBCL cell line SU-DHL4 was obtained from Dr. Lee Honigberg (Pharmacyclics Inc., Sunnyvale, CA).

Techniques: In Vitro, Cell Culture, Staining

Journal: Leukemia & lymphoma

Article Title: CD40 ligand is necessary and sufficient to support primary diffuse large B-cell lymphoma cells in culture: a tool for in vitro preclinical studies with primary B-cell malignancies

doi: 10.3109/10428194.2011.654337

Figure Lengend Snippet: Phenotypic characteristics of DLBCL cells cultured with KtCD40L. (A) DLBCL cells cultured with KtCD40L for 48 days were phenotyped using flow cytometry. Gray lines represent unstained samples. One representative sample of three DLBCL cases analyzed is shown. (B) Clonal IgH gene rearrangements of the same size (105 bp) were observed in tumor cells before (Day 0) and after (Day 48) the culture. One representative sample of two DLBCL cases analyzed is shown. (C) Examples of FISH images obtained from one DLBCL case with trisomy of CFA 31. Control dog metaphase chromosomes (a) and an interphase nucleus (b) probed with a marker for CFA 31 (yellow signal), exhibiting the expected chromosomal location and normal copy number status (n = 2) of this region. Three copies of this locus were evident in 51% of DLBCL cells prior to culture (c), and in 48% of DLBCL cells after culture with KtCD40L for 40 days (d). A KtCD40L cell (upper left without a probe signal) and a DLBCL cell (lower right with probe signals) are shown together in (d).

Article Snippet: The human DLBCL cell line SU-DHL4 was obtained from Dr. Lee Honigberg (Pharmacyclics Inc., Sunnyvale, CA).

Techniques: Cell Culture, Flow Cytometry, Control, Marker

Journal: Leukemia & lymphoma

Article Title: CD40 ligand is necessary and sufficient to support primary diffuse large B-cell lymphoma cells in culture: a tool for in vitro preclinical studies with primary B-cell malignancies

doi: 10.3109/10428194.2011.654337

Figure Lengend Snippet: Feeder cell-free culture of primary DLBCL cells with recombinant shuCD40L. (A) The MTS cell proliferation assay was performed at 24-hour intervals on primary dog DLBCL cells stimulated with increasing amounts of shuCD40L. Data from 4 independent primary dog DLBCL samples are shown. Data were normalized to results from 5 × 104 cells (0 hour) to show fold increase of cell numbers during the culture period. (B) MTS cell proliferation assay was performed on primary DLBCL cells stimulated with 100 ng/mL shuCD40L in the presence of increasing amounts of anti-human CD40L-IgA. Three different primary DLBCL samples were tested for 72 hours in duplicate conditions and data were normalized to results of cells cultured with 100 ng/mL shuCD40L without neutralizing antibodies. Data shown represent the mean ± SD (n = 3) and statistical significance was tested against the condition of 100 ng/mL shuCD40L without neutralizing antibodies using ANOVA (*P < 0.01, **P < 0.001). (C) Human B-ALL samples were stained for CD22 and CD40. CD22+ tumor cells (> 90% CD22+ cells in both samples) were gated and CD40 expression was analyzed. Gray lines represent staining with an isotype control antibody. (D) The MTS cell proliferation assay using primary human B-ALL cells was performed as described above. Data from 2 independent samples are shown.

Article Snippet: The human DLBCL cell line SU-DHL4 was obtained from Dr. Lee Honigberg (Pharmacyclics Inc., Sunnyvale, CA).

Techniques: Recombinant, Proliferation Assay, Cell Culture, Staining, Expressing, Control

Journal: Leukemia & lymphoma

Article Title: CD40 ligand is necessary and sufficient to support primary diffuse large B-cell lymphoma cells in culture: a tool for in vitro preclinical studies with primary B-cell malignancies

doi: 10.3109/10428194.2011.654337

Figure Lengend Snippet: IC 50 of 22KDEL and Bic3 for tumor cells

Article Snippet: The human DLBCL cell line SU-DHL4 was obtained from Dr. Lee Honigberg (Pharmacyclics Inc., Sunnyvale, CA).

Techniques:

Journal: Leukemia & lymphoma

Article Title: CD40 ligand is necessary and sufficient to support primary diffuse large B-cell lymphoma cells in culture: a tool for in vitro preclinical studies with primary B-cell malignancies

doi: 10.3109/10428194.2011.654337

Figure Lengend Snippet: Comparable assessment of cytotoxicity using dog and human B-cell malignancies. (A) Four independent primary dog DLBCL samples; #1–3 are the same as in Figure 4, #4 is from a different dog, and (B) two human primary B-ALL samples from different patients than those shown in Figure 4, were cultured for 72 hours with a log serial dilutions (0.01–100 nM) of 22KDEL and Bic3 immunotoxins in the presence of shuCD40L. Experiments were performed in duplicate and cell numbers were determined using the MTS assay. Results with 10 nM immunotoxins are shown, and a summary result of the IC50 for 22KDEL and Bic3 is shown in Table I.

Article Snippet: The human DLBCL cell line SU-DHL4 was obtained from Dr. Lee Honigberg (Pharmacyclics Inc., Sunnyvale, CA).

Techniques: Cell Culture, MTS Assay

Journal: Translational Oncology

Article Title: Melatonin increases the chemosensitivity of diffuse large B-cell lymphoma cells to epirubicin by inhibiting P-glycoprotein expression via the NF-κB pathway

doi: 10.1016/j.tranon.2020.100876

Figure Lengend Snippet: Effect of melatonin (MLT) and epirubicin (EPI) combination of DLBCL cells proliferation. (A) Human SUDHL-10 and SUDHL-6 cells were treated with MLT or EPI alone or their combination at the indicated doses. At 48 h after treatment, the cell viability was determined by a CCK8 assay. The cells treated with vehicle control DMSO were used as the referent group with cell viability set at 100%. (B) The IC50 values of EPI for cell viability inhibition in cells treated with or without MLT determined. The data were presented as mean ± SD of three separate experiments. The level of significant was indicated by * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The human DLBCL cell lines SUDHL-10 and SUDHL-6 were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: CCK-8 Assay, Control, Inhibition

Journal: Translational Oncology

Article Title: Melatonin increases the chemosensitivity of diffuse large B-cell lymphoma cells to epirubicin by inhibiting P-glycoprotein expression via the NF-κB pathway

doi: 10.1016/j.tranon.2020.100876

Figure Lengend Snippet: Melatonin suppressed the function of P-glycoprotein in DLBCL cells. SUDHL-10 and SUDHL-6 cells were treated with MLT or RPMI 1640 (control) for 48 h. Then cells were incubated with 1 μg/mL Rhodamine-123 (Rho-123) dye or EPI for 90 min in dark. (A) The effect of MLT on intracellular Rho-123 accumulation in SUDHL-10 and SUDHL-6 cells was measured by flow cytometry. (B) Quantitatively analysis of intracellular Rho-123 fluorescence. (C) The effect of MLT on intracellular EPI accumulation in SUDHL-10 and SUDHL-6 cells was measured by flow cytometry. (D) Quantitatively analysis of intracellular EPI fluorescence. Each column shows the mean ± SD of three independent experiments. (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: The human DLBCL cell lines SUDHL-10 and SUDHL-6 were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Control, Incubation, Flow Cytometry, Fluorescence

Journal: Translational Oncology

Article Title: Melatonin increases the chemosensitivity of diffuse large B-cell lymphoma cells to epirubicin by inhibiting P-glycoprotein expression via the NF-κB pathway

doi: 10.1016/j.tranon.2020.100876

Figure Lengend Snippet: Epirubicin induced the up-regulation of P-glycoprotein expression by activating the NF-κB signaling pathway. (A) HE staining and IHC analysis of P-gp and P65 in DLBCL tissue; representative pictures of each antibody staining are shown. (B) Scatterplot showing the positive correlation between P-gp and P65 expression in patients; Spearman's coefficient tests were performed to assess statistical significance. (C) P-gp and P65 protein expression were determined by Western blotting after 48 h treatment with EPI at various indicated concentrations. (D) Cells were treated with or without NF-κB-selective inhibitor QNZ (500 nM) for 8 h after pretreatment with a higher concentration of EPI (100 ng/mL) for 48 h. P-gp and P65 protein expression were determined by Western blotting.

Article Snippet: The human DLBCL cell lines SUDHL-10 and SUDHL-6 were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Expressing, Staining, Paraffin-embedded Immunohistochemistry, Western Blot, Concentration Assay

Journal: Translational Oncology

Article Title: Melatonin increases the chemosensitivity of diffuse large B-cell lymphoma cells to epirubicin by inhibiting P-glycoprotein expression via the NF-κB pathway

doi: 10.1016/j.tranon.2020.100876

Figure Lengend Snippet: The schematic diagram of the molecular mechanisms of melatonin enhancing the sensitivity of DLBCL cells to epirubicin. The co-treatment of epirubicin and melatonin increased proliferation suppression and apoptosis induction might be achieved through activating the cytochrome c /caspase-dependent apoptotic signaling and inhibiting the NF-κB signaling. The red symbol (┤) indicates negative regulation. Black colored arrow (→) indicates direct or indirect positive regulation.

Article Snippet: The human DLBCL cell lines SUDHL-10 and SUDHL-6 were obtained from the China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: