Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A diphtheria toxin-based nanoparticle achieves specific cytotoxic effect on CXCR4 + lymphoma cells without toxicity in immunocompromised and immunocompetent mice.

doi: 10.1016/j.biopha.2022.112940

Figure Lengend Snippet: Fig. 1. T22-DITOX-H6 specific cytotoxicity in human CXCR4+ DLBCL cell lines. (A) Cell viability assays performed after 48 h of T22-DITOX-H6 incubation at different concentrations (0.05–5.00 nM) in Toledo and U-2932 cells. (B) Competition assays in Toledo and U-2932 cells with or without pretreating with the antagonist of CXCR4 AMD3100 followed by the nanoparticle incubation for 48 h (ratio 10 AMD3100: 1 T22-DITOX-H6). (C-D) CXCR4 determination in SUDHL-2 and CXCR4+ SUDHL-2 cells by flow cytometry and Western blot. In panel D, α/β tubulin antibody is used as endogen control. (E) Cell viability assays performed in SUDHL-2 and CXCR4+ SUDHL-2 cells after 48 h of T22-DITOX-H6 incubation (0.10–5.00 nM). *p ≤0.05.

Article Snippet: Then, membranes containing Toledo and U-2932 cell lysates were incubated with the following primary antibodies: 1:2000 PARP (556494, BD Biosciences), 1:1000 caspase-3 (610322, BD Biosciences) and 1:1000 cleaved caspase-3 (9661, Cell Signaling Technology, Danvers, MA, USA); SUDHL-2 and CXCR4+ SUDHL-2 membranes were incubated with 1:1000 CXCR4 (1009, Prosci, Fort Collins, CO, USA).

Techniques: Incubation, Flow Cytometry, Western Blot, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A diphtheria toxin-based nanoparticle achieves specific cytotoxic effect on CXCR4 + lymphoma cells without toxicity in immunocompromised and immunocompetent mice.

doi: 10.1016/j.biopha.2022.112940

Figure Lengend Snippet: Fig. 2. Cell death mechanism induced by T22-DITOX-H6 in human CXCR4+ DLBCL cell lines. (A) Cell viability assays, pretreating Toledo and U-2932 cells with 100 μM of z-VAD-FMK pan caspase inhibitor, followed by the incubation of 5 nM of T22-DITOX-H6 for 48 h. (B) Western Blot analysis for the detection of pro- caspase-3, cleaved-caspase-3, full length Poly (ADP-ribose) polymerase (PARP) and cleaved PARP in Toledo and U-2932 cells, treated with buffer or 5 nM T22- DITOX-H6 for 24 h and 48 h. α/β tubulin antibody is used as endogen control. (C) Representative dot-plot from Annexin-PI assay showing the percent of viable cells (V), early apoptotic cells (EA) and late apoptotic cells (LA) in Toledo an U-2932 cells treated with buffer or 5 nM T22-DITOX-H6 at different time points (15 h, 24 h and 48 h) and (D) their respective quantification in triplicates. (E) DAPI staining images (1000X) of Toledo and U-2932 cells treated with buffer or 2.5 nM T22- DITOX-H6 for 48 h. White arrows point out apoptotic bodies. *p ≤0.05.

Article Snippet: Then, membranes containing Toledo and U-2932 cell lysates were incubated with the following primary antibodies: 1:2000 PARP (556494, BD Biosciences), 1:1000 caspase-3 (610322, BD Biosciences) and 1:1000 cleaved caspase-3 (9661, Cell Signaling Technology, Danvers, MA, USA); SUDHL-2 and CXCR4+ SUDHL-2 membranes were incubated with 1:1000 CXCR4 (1009, Prosci, Fort Collins, CO, USA).

Techniques: Incubation, Western Blot, Control, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A diphtheria toxin-based nanoparticle achieves specific cytotoxic effect on CXCR4 + lymphoma cells without toxicity in immunocompromised and immunocompetent mice.

doi: 10.1016/j.biopha.2022.112940

Figure Lengend Snippet: Fig. 3. In vivo antineoplastic effect of T22-DITOX-H6 in a disseminated CXCR4+ DLBCL mouse model. (A) BLI emitted by Toledo-Luci cells from four representative mice of each group (buffer and T22-DITOX-H6) at days 28, 30 and 33. (B) BLI quantification of mice treated with buffer or T22-DITOX-H6 during the follow-up (n = 9/group). (C) Body weight of mice treated with buffer or T22-DITOX-H6 during the follow-up (n = 9/group). *** p < 0.005. BLI: bioluminescence imaging; iv: intravenous.

Article Snippet: Then, membranes containing Toledo and U-2932 cell lysates were incubated with the following primary antibodies: 1:2000 PARP (556494, BD Biosciences), 1:1000 caspase-3 (610322, BD Biosciences) and 1:1000 cleaved caspase-3 (9661, Cell Signaling Technology, Danvers, MA, USA); SUDHL-2 and CXCR4+ SUDHL-2 membranes were incubated with 1:1000 CXCR4 (1009, Prosci, Fort Collins, CO, USA).

Techniques: In Vivo, Imaging

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A diphtheria toxin-based nanoparticle achieves specific cytotoxic effect on CXCR4 + lymphoma cells without toxicity in immunocompromised and immunocompetent mice.

doi: 10.1016/j.biopha.2022.112940

Figure Lengend Snippet: Fig. 4. Ex vivo antineoplastic effect of T22-DITOX-H6 in a disseminated CXCR4+ DLBCL mouse model. BLI quantification and representative images of lymphoma- infiltrated organs (cervical LNs, renal LNs, backbone, hindlimbs and cranium) from Toledo-Luci mice treated either with buffer or T22-DITOX-H6 at the end of the experiment. *p ≤0.05, ***p ≤0.005. BLI: bioluminescence imaging; LNs: lymph nodes.

Article Snippet: Then, membranes containing Toledo and U-2932 cell lysates were incubated with the following primary antibodies: 1:2000 PARP (556494, BD Biosciences), 1:1000 caspase-3 (610322, BD Biosciences) and 1:1000 cleaved caspase-3 (9661, Cell Signaling Technology, Danvers, MA, USA); SUDHL-2 and CXCR4+ SUDHL-2 membranes were incubated with 1:1000 CXCR4 (1009, Prosci, Fort Collins, CO, USA).

Techniques: Ex Vivo, Imaging

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A diphtheria toxin-based nanoparticle achieves specific cytotoxic effect on CXCR4 + lymphoma cells without toxicity in immunocompromised and immunocompetent mice.

doi: 10.1016/j.biopha.2022.112940

Figure Lengend Snippet: Fig. 5. T22-DITOX-H6 specific cytotoxicity in a murine CXCR4+ lymphoma cell line. (A-B) CXCR4 determination by immunohistochemistry and flow cytometry in murine A-20 cells. Scale bars: main picture at 50 µm (400X) and the inset at 20 µm (1000X). (C) Cell viability assays measuring the T22-DITOX-H6 cytotoxicity in A- 20 cells at different concentrations of the nanoparticle (0.25–5.00 nM) for 48 h. (D) Competition assays in A-20 cells with or without the pretreatment of the CXCR4 antagonist AMD3100 followed by the nanoparticle incubation for 48 h (ratio 10 AMD3100: 1 T22-DITOX-H6). MFI: mean fluorescence intensity. (E) DAPI staining pictures (1000X) of A-20 cells treated with buffer or 5 nM T22-DITOX-H6 for 48 h. White arrows point out apoptotic bodies. *p ≤0.05.

Article Snippet: Then, membranes containing Toledo and U-2932 cell lysates were incubated with the following primary antibodies: 1:2000 PARP (556494, BD Biosciences), 1:1000 caspase-3 (610322, BD Biosciences) and 1:1000 cleaved caspase-3 (9661, Cell Signaling Technology, Danvers, MA, USA); SUDHL-2 and CXCR4+ SUDHL-2 membranes were incubated with 1:1000 CXCR4 (1009, Prosci, Fort Collins, CO, USA).

Techniques: Immunohistochemistry, Flow Cytometry, Incubation, Fluorescence, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: A diphtheria toxin-based nanoparticle achieves specific cytotoxic effect on CXCR4 + lymphoma cells without toxicity in immunocompromised and immunocompetent mice.

doi: 10.1016/j.biopha.2022.112940

Figure Lengend Snippet: Fig. 6. T22-DITOX-H6 anti-tumor effect, and on-target and off-target toxicity in a SC mouse model bearing murine CXCR4+ lymphoma cells. (A) A-20 SC tumor volume from mice treated with buffer (n = 5) or T22-DITOX-H6 (n = 4) during the whole experiment. (B) AUC quantification of tumor volume from 14 to 20 days in buffer and nanoparticle-treated mice. (C) Body weight follow-up of mice bearing A-20 SC tumors, treated with buffer and T22-DITOX-H6. (D) Hematoxylin and eosin staining from on target and off-target organs (spleen, liver, kidneys and BM). Scale bars: 50 µm. *p ≤0.05. AUC: area under the curve; BM: bone marrow; iv: intravenous.

Article Snippet: Then, membranes containing Toledo and U-2932 cell lysates were incubated with the following primary antibodies: 1:2000 PARP (556494, BD Biosciences), 1:1000 caspase-3 (610322, BD Biosciences) and 1:1000 cleaved caspase-3 (9661, Cell Signaling Technology, Danvers, MA, USA); SUDHL-2 and CXCR4+ SUDHL-2 membranes were incubated with 1:1000 CXCR4 (1009, Prosci, Fort Collins, CO, USA).

Techniques: Staining

Journal: World journal of gastroenterology

Article Title: CXCR4/SDF-1 axis is involved in lymph node metastasis of gastric carcinoma.

doi: 10.3748/wjg.v17.i19.2389

Figure Lengend Snippet: Figure 4 Expression of CXC chemokine receptor-4 in gastric carcinoma tissues and stromal cell-derived factor-1 in lymph nodes. A: CXC chemokine recep- tor-4 (CXCR4) protein was detected by immunohistochemistry in primary gastric carcinoma tissues; B: CXCR4 protein was not detected in normal mucous membrane; C: Stromal cell-derived factor-1 (SDF-1) protein was detected by immunohistochemistry in lymph nodes with gastric cancer cell metastasis; D: SDF-1 protein was not detected in normal lymph nodes (400 ×).

Article Snippet: Subsequently, the slides were pretreated with 1% bovine serum albumin in phosphate-buffered saline (PBS) and incubated with antiSDF-1 antibody (Boster; dilution 1:100) and anti-CXCR4 antibody (Boster; dilution 1:50) for 1 h at room temperature.

Techniques: Expressing, Derivative Assay, Immunohistochemistry, Membrane

Journal: World journal of gastroenterology

Article Title: CXCR4/SDF-1 axis is involved in lymph node metastasis of gastric carcinoma.

doi: 10.3748/wjg.v17.i19.2389

Figure Lengend Snippet: Figure 5 mRNA expression of CXC chemokine receptor-4 in gastric can- cer cells, tumors and normal mucous membranes and of stromal cell- derived factor-1 in lymph nodes. A: CXCR

Article Snippet: Subsequently, the slides were pretreated with 1% bovine serum albumin in phosphate-buffered saline (PBS) and incubated with antiSDF-1 antibody (Boster; dilution 1:100) and anti-CXCR4 antibody (Boster; dilution 1:50) for 1 h at room temperature.

Techniques: Expressing, Derivative Assay

Journal: Cell

Article Title: Mobilization-based chemotherapy-free engraftment of gene-edited human hematopoietic stem cells

doi: 10.1016/j.cell.2022.04.039

Figure Lengend Snippet:

Article Snippet: Anti-human CD184 , Miltenyi Biotec , Cat# 130-117-519; RRID:AB_2734059.

Techniques: Blocking Assay, Plasmid Preparation, Recombinant, Electroporation, CRISPR, Adjuvant, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Cell Culture, Gene Expression

Journal:

Article Title: Bicyclams, Selective Antagonists of the Human Chemokine Receptor CXCR4, Potently Inhibit Feline Immunodeficiency Virus Replication

doi:

Figure Lengend Snippet: Inhibition of binding of the anti-CXCR4 MAb to feline thymocytes in the presence (C) or absence (B) of AMD3100 at 1 μg/ml. In panel A, results obtained with an isotype control MAb are shown. The percentages of fluorescence-positive cells and the mean fluorescence intensity values are indicated in each histogram.

Article Snippet: AMD3100 had no inhibitory effect (at 100 μg/ml) on the binding of feline anti-CD4 MAb (MCA 1347; Serotec, Oxford, United Kingdom) and anti-CD8 MAb (MCA 1350; Serotec) in feline thymocytes. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 4 caption a7 Inhibition of binding of the anti-CXCR4 MAb to feline thymocytes in the presence (C) or absence (B) of AMD3100 at 1 μg/ml.

Techniques: Inhibition, Binding Assay, Fluorescence