Journal: Scientific Reports

Article Title: The oncolytic adenovirus VCN-01 promotes anti-tumor effect in primitive neuroectodermal tumor models

doi: 10.1038/s41598-019-51014-1

Figure Lengend Snippet: Characterization of VCN-01 in PNET cell lines in vitro . ( A ) Expression of adenoviral receptors CAR, α v β 3 integrin and α v β 5 integrin in CNS-PNET cell lines PFSK-1 (left) and SK-PN-DW (right). Graph shows the percentage of stained cells for each receptor (Mean ± SD; n = 3). ( B ) PFSK-1 and SK-PN-DW (200,000 cells) infected with the GFP expressing vector AdTLRGDK at MOIs of 0, 0.1, 1, 10 or 100 PFU/cell. Graph indicates the percentage of GFP positive cells expression measured by flow cytometry at 24 h (white bars) or 48 h (black bars) after the infection (Mean ± SD; n = 3). ( C ) Detection of the viral proteins E1A and fiber in whole-cell lysates 48 h after being from PFSK-1 and SK-PN-DW from VCN-01 infected PFSK-1 and SK-PN-DW cultures. Grb2 was used as loading control protein. Blots from different parts of the same gel have been grouped to improve clarity of the image. ( D ) Viral titers in PFSK-1 and SK-PN-DW (50,000 cells/well) cultures 72 h after being infected with VCN-01 at MOIs 1 and 10. Bars represent total PFUs contained in the lysates (Mean ± SD; n = 3).

Article Snippet: PNET cell lines PFSK-1 (ATCC, Manassas, VA, USA; CRL-2060 TM ) and SK-PN-DW (ATCC, CRL-2139 TM ) were stained with unlabeled monoclonal antibodies recognizing the adenoviral receptors CAR (Merck Millipore, Temecula, CA, USA; 05–644) α v β 3 integrin (Merck Millipore; CBL544) and α v β 5 integrin (R&D Systems Inc., Minneapolis, MN, USA; MAB2528), and donkey anti-mouse IgG1-AlexaFluor488 (Thermo Fisher Scientific, Waltham, MA, USA; A-21202) as secondary antibody.

Techniques: In Vitro, Expressing, Staining, Infection, Plasmid Preparation, Flow Cytometry, Control

Journal: Biomolecules

Article Title: Macrophage-like THP-1 Cells Derived from High-Density Cell Culture Are Resistant to TRAIL-Induced Cell Death via Down-Regulation of Death-Receptors DR4 and DR5

doi: 10.3390/biom12020150

Figure Lengend Snippet: Micrographs of THP-1ad cells after 1 day of incubation with 20 μM of Cilengitide and without Cilengitide (control); staining of the cytoplasm with Calcein AM (green), and of cell nuclei with Hoechst 33342 (blue) ( a ). Percentage of THP-1ad cells attached to the surface of culture dishes 1 day after incubation with anti-αVβ3 and anti-αVβ5 antibodies, with isotypic control Ig G1, and Cilengitide relative to control (90% of attached cells) ( b ). *— p < 0.05 in comparison with control.

Article Snippet: THP-1ad cells were seeded in the amount of 3 × 10 4 per well of a 96-well plate in 100 μL of growth medium supplemented with anti-αVβ3 monoclonal antibody (ab78289, Abcam, Cambridge, UK) or anti-αVβ5 monoclonal antibody (MAB2528, R&DSystems Minneapolis, MN, USA), or with isotypic control to these antibodies (FITC anti-mouse IgG1, Biolegend, San Diego, CA, USA) at a concentration of 17 μg/mL, or with Cilengitide (Selleckchem, Pittsburgh, PA, USA) [ ] at concentration of 20 μM.

Techniques: Incubation, Staining

Journal: Cancer Science

Article Title: Myeloid suppressor cell‐associated immune dysfunction in CSA1M fibrosarcoma tumor‐bearing mice

doi: 10.1111/j.1349-7006.2007.00465.x

Figure Lengend Snippet: Tumor burden caused splenomegaly and altered cell percentages in spleen. Male BALB/c mice were inoculated with CSA1M tumor cells as described in Materials and Methods. Eight to 10 weeks after cell inoculation, mice were killed and spleens were harvested. (a) The numbers of splenocytes were determined by trypan blue exclusion assay. (b) Splenocytes were stained for CD4+, CD8+, B220+, or Mac‐1+ and analyzed with flow cytometry. Data are expressed as mean ± standard error of the mean (SEM) of three independent experiments that give similar results. *P < 0.05, **P < 0.01, ***P < 0.001 versus normal control.

Article Snippet: CSA1M fibrosarcoma, ( 12 ) which was induced in BALB/c mouse with the Schmidt‐Ruppin strain of Rous sarcoma virus and shown to produce no Rous sarcoma virus, was kindly provided by Dr T. Yoshida, Hamamatsu Medical College, Hamamatsu, Japan.

Techniques: Trypan Blue Exclusion Assay, Staining, Flow Cytometry, Control

Journal: Cancer Science

Article Title: Myeloid suppressor cell‐associated immune dysfunction in CSA1M fibrosarcoma tumor‐bearing mice

doi: 10.1111/j.1349-7006.2007.00465.x

Figure Lengend Snippet: Tumor growth did not affect T cell function in vitro but impaired it in vivo. (a) Splenic CD4+ or CD8+ T cells (3 × 106/mL) from normal mice or mice at late CSA1M tumor‐bearing stage were cultured without or with ConA (5 µg/mL) for 48 h and pulsed with [3H]‐thymidine. The incorporated radioactivity was measured. Data are mean ± standard deviation (SD) of triplicate cultures. (b) Splenic CD4+ or CD8+ T cells (3 × 106/mL) were cultured without or with ConA (5 µg/mL) for 24 h and culture supernatants were assayed for interferon‐γ (IFN‐γ) and interleukin‐4 (IL‐4) by enzyme‐linked immunosorbent assay (ELISA). (c) 2,4‐Dinitrofluorobenzene (DNFB)‐induced ear swelling. Mice were initially sensitized with DNFB on days 0 and 1, and then challenged with DNFB on day 9. Ear swelling was calculated as the difference between the weights of left (DNFB‐treated) and right (untreated) ear punches 30 h after challenge. Data are expressed as mean ± standard deviation (SD) n = 8 mice/group. Three independent experiments were carried out that give similar results. *P < 0.001 versus normal control.

Article Snippet: CSA1M fibrosarcoma, ( 12 ) which was induced in BALB/c mouse with the Schmidt‐Ruppin strain of Rous sarcoma virus and shown to produce no Rous sarcoma virus, was kindly provided by Dr T. Yoshida, Hamamatsu Medical College, Hamamatsu, Japan.

Techniques: Cell Function Assay, In Vitro, In Vivo, Cell Culture, Radioactivity, Standard Deviation, Enzyme-linked Immunosorbent Assay, Control