Journal: International Journal of Molecular Sciences

Article Title: Roles of OX40 and OX40 Ligand in Mycosis Fungoides and Sézary Syndrome

doi: 10.3390/ijms222212576

Figure Lengend Snippet: OX40 and OX40 ligand (OX40L) expression in patients with cutaneous T-cell lymphoma (CTCL). ( A , B ) Quantitative RT-PCR was performed to measure expression levels of OX40 and OX40L using mRNA extracted from MF/SS lesional skin ( n = 57; 49 MF cases and 8 SS cases) and normal skin ( n = 13). ( C ) Correlations between soluble IL-2 receptor (sIL-2R) and OX40 or OX40L mRNA expression in MF/SS patients. ( D ) Positive correlations between OX40 and OX40L mRNA expression in MF/SS lesional skin. ( E ) Serum soluble OX40 and soluble OX40L levels in CTCL patients. ( F ) MF/SS patients with high OX40 mRNA expression exhibited poor prognosis. Means are presented as bars. ** p < 0.01.

Article Snippet: Anti-human OX40 (2 μg/mL; AF3388; R&D Systems) and anti-human OX40L (2 μg/mL; clone MM0505-8S23; Novus Biologicals, Centennial, CO, USA) were used as neutralizing antibodies.

Techniques: Expressing, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Roles of OX40 and OX40 Ligand in Mycosis Fungoides and Sézary Syndrome

doi: 10.3390/ijms222212576

Figure Lengend Snippet: OX40 and OX40L expression in lesional skin of MF/SS was determined by immunohistochemistry (original magnification ×400, scale bar = 100 µm). ( A ) Representative results are shown. ( B ) Counts of OX40 and OX40L-positive cells. Ten cases of patch/plaque MF, nine cases of tumor MF/SS, and seven cases of healthy subjects. ** p < 0.01.

Article Snippet: Anti-human OX40 (2 μg/mL; AF3388; R&D Systems) and anti-human OX40L (2 μg/mL; clone MM0505-8S23; Novus Biologicals, Centennial, CO, USA) were used as neutralizing antibodies.

Techniques: Expressing, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: Roles of OX40 and OX40 Ligand in Mycosis Fungoides and Sézary Syndrome

doi: 10.3390/ijms222212576

Figure Lengend Snippet: OX40 and OX40L expression was analyzed by flow cytometry in CD4 + CD7 − T cells from 6 Sézary syndrome (SS) patients and CD4 + T cells from 6 healthy controls. Comparison of OX40 and OX40L-positive cell ratios in peripheral blood CD4 + CD7 − T cells in patients with SS or in healthy controls peripheral blood CD4 + T cells. n.s.: not significant. * p < 0.05.

Article Snippet: Anti-human OX40 (2 μg/mL; AF3388; R&D Systems) and anti-human OX40L (2 μg/mL; clone MM0505-8S23; Novus Biologicals, Centennial, CO, USA) were used as neutralizing antibodies.

Techniques: Expressing, Flow Cytometry, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Roles of OX40 and OX40 Ligand in Mycosis Fungoides and Sézary Syndrome

doi: 10.3390/ijms222212576

Figure Lengend Snippet: Involvement of OX40 and OX40L in CTCL cell lines. ( A ) OX40 and OX40L expression was analyzed by flow cytometry in human CTCL cell lines (Hut78, MyLa, and HH cells). ( B ) HH, Hut, and MyLa cells (1.0 × 10 5 /well) were cultured with anti-OX40 (5 μg/mL) and/or anti-OX40L (5 μg/mL) neutralizing antibodies for 48 hours. ( C ) Hut and HH cells (1.0 × 10 5 /well) were cultured with anti-OX40 neutralizing antibody (5 μg/mL) for 24 hours. Apoptosis (Annexin V + and 7-AAD − ) was evaluated with Annexin V and 7-AAD staining for flow cytometric analysis. * p < 0.05, ** p < 0.01.

Article Snippet: Anti-human OX40 (2 μg/mL; AF3388; R&D Systems) and anti-human OX40L (2 μg/mL; clone MM0505-8S23; Novus Biologicals, Centennial, CO, USA) were used as neutralizing antibodies.

Techniques: Expressing, Flow Cytometry, Cell Culture, Staining

Journal: International Journal of Molecular Sciences

Article Title: Roles of OX40 and OX40 Ligand in Mycosis Fungoides and Sézary Syndrome

doi: 10.3390/ijms222212576

Figure Lengend Snippet: Anti-OX40 and anti-OX40L antibodies suppressed tumor growth. Hut78 cells (5.0 × 10 6 cells) in 100 µL of PBS were injected subcutaneously into the shaved left abdomen of NSG mice. Anti-OX40 or anti-OX40L neutralizing antibodies were injected on days 0, 4, 7, and 11 intraperitoneally. ( A ) Representative images are shown. ( B ) The tumor volume was calculated using the equation: V = π (L1 × L2 2 )/6, where V = volume (mm 3 ), L1 = longest diameter (mm), and L2 = shortest diameter (mm). Values are means ± SEM. *,# p < 0.05 by Mann–Whitney U test compared with control group.

Article Snippet: Anti-human OX40 (2 μg/mL; AF3388; R&D Systems) and anti-human OX40L (2 μg/mL; clone MM0505-8S23; Novus Biologicals, Centennial, CO, USA) were used as neutralizing antibodies.

Techniques: Injection, MANN-WHITNEY, Control

Journal: Retrovirology

Article Title: Increased expression of OX40 is associated with progressive disease in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/1742-4690-10-51

Figure Lengend Snippet: Tax-dependent constitutive expression of OX40 in HTLV-1-infected T-cell lines and Tax-inducible JPX-9 cell line. A. Representative histograms of OX40 expression in 6 HTLV-1 infected T-cell lines (HUT-102, MT-1, MT-2, MT-4, C5/MJ, SLB-1) and two HTLV-1-uninfected T-cell lines (CEM-OX40L and CEM-OX40). Shaded histograms represent the isotype control. Tax+ or Tax- means whether these cells express Tax (Tax+) or not (Tax-). B. Representative histograms of OX40L expression in 6 HTLV-1 infected T-cell lines (HUT-102, MT-1, MT-2, MT-4, C5/MJ, SLB-1) and two HTLV-1-uninfected T-cell lines (CEM-OX40L and CEM-OX40). Shaded histograms are isotype controls. C. Flow cytometric analysis of expression of OX40 after induction of Tax in JPX-9 cells. D. Flow cytometric analysis of expression of OX40L after induction of Tax in JPX-9 cells. E. Soluble OX40 and OX40L levels in cell culture supernatant and cell lysate from 6 HTLV-1 infected T-cell lines (HUT-102, MT-1, MT-2, MT-4, C5/MJ, SLB-1) and three HTLV-1-uninfected T-cell lines (CEM-mock, CEM-OX40L and CEM-OX40). F. Soluble OX40 and OX40L levels in cell culture supernatant and cell lysate from JPX-9 cell line treated with CdCl 2 along with the induction of viral transactivator Tax.

Article Snippet: Recombinant human OX40 ligand/TNFSF4 and recombinant human OX40/TNFRSF4/Fc Chimera were purchased from R&D Systems (Minneapolis, MN) and used for the standard curve in sOX40L and sOX40 ELISA, respectively.

Techniques: Expressing, Infection, Cell Culture

Journal: Retrovirology

Article Title: Increased expression of OX40 is associated with progressive disease in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/1742-4690-10-51

Figure Lengend Snippet: OX40 is specifically expressed on the surface of T cells naturally infected with HTLV-1 that have the potential to produce pro-inflammatory cytokines. A. OX40 was detected on CD4 + T cells of HAM/TSP patient with anti-OX40 mAb (clones B-7B5) after 16 hours in vitro cultivation in the absence of any growth factors or mitogen. B. OX40L was not detected on CD4 + T cells of HAM/TSP patient with anti-OX40L mAb (clones 5A8) after 16 hours in vitro cultivation in the absence of any growth factors or mitogen. C. Tax protein was detected in CD4 + T cells of HAM/TSP patient after 16 hours in vitro cultivation. D. OX40 was expressed almost exclusively in naturally infected CD4 + T cells that also expressed Tax in HAM/TSP patient. E. Both HTLV-1 tax and OX40 mRNA expression in CD4 + T cells was increased after 16 hours in vitro cultivation. F. The frequency of pro-inflammatory cytokine positive cells within the OX40 + CD4 + and Tax + CD4 + populations from HTLV-1 infected individuals are significantly higher than OX40 - CD4 + and Tax - CD4 + T cells, respectively (p<0.001, Student’s t- test). One representative experiment of HAM/TSP patient (HAM/TSP1) is shown.

Article Snippet: Recombinant human OX40 ligand/TNFSF4 and recombinant human OX40/TNFRSF4/Fc Chimera were purchased from R&D Systems (Minneapolis, MN) and used for the standard curve in sOX40L and sOX40 ELISA, respectively.

Techniques: Infection, Clone Assay, In Vitro, Expressing

Journal: Retrovirology

Article Title: Increased expression of OX40 is associated with progressive disease in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/1742-4690-10-51

Figure Lengend Snippet: Increased expression of OX40 in vivo in rapidly progressive HAM/TSP patients. A. The plasma levels of soluble OX40 (sOX40) measured by ELISA. The plasma levels of sOX40 in typical HAM/TSP patients (chronic HAM: n=20), asymptomatic carriers (ACs: n=9) and normal uninfected healthy controls (NCs: n=13). B. No correlation between the plasma levels of sOX40 and HTLV-1 proviral load (tax copies/10,000PBMCs) from 29 HTLV-1 infected individuals (20 chronic HAM/TSP patients and 9 ACs). Data were analyzed by Spearman rank correlation. C. The cerebrospinal fluid (CSF) levels of sOX40 in rapidly progressive HAM/TSP patients (n=3), chronic HAM/TSP patients (n=22) and other neurological diseases including multiple sclerosis (MS) (n=12), aseptic meningitis (n=8), systemic lupus erythematosus (SLE) with neurological manifestations (n=5), chronic inflammatory demyelinating polyneuropathy (CIDP) (n=9), Guillain-Barré syndrome (GBS) (n=6), and amyotrophic lateral sclerosis (ALS) (n=9). Chronic HAM/TSP means typical cases fulfilling diagnostic criteria and rapidly progressive HAM/TSP is defined by patients’ incapacity to walk unaided within three months after symptoms’ onset. D. The levels of sOX40 in the CSF from HTLV-1 infected other inflammatory neurological diseases (HTLV-1+ OINDs), i.e. any inflammatory neurological disorders except for HAM/TSP which occurred in HTLV-1 infected individuals, was not significantly different from that of chronic HAM/TSP, whereas the levels of sOX40 from HTLV-1+ OINDs was significantly increased than that of non-infected OINDs (HTLV-1- OINDs). HTLV-1+ OINDs: 1 multiple sclerosis (MS), 1 SLE with neurological manifestations, 4 aseptic meningitis. HTLV-1- OINDs: 9 MS, 5 SLE with neurological manifestations, 7 CIDP, 5 GBS.

Article Snippet: Recombinant human OX40 ligand/TNFSF4 and recombinant human OX40/TNFRSF4/Fc Chimera were purchased from R&D Systems (Minneapolis, MN) and used for the standard curve in sOX40L and sOX40 ELISA, respectively.

Techniques: Expressing, In Vivo, Enzyme-linked Immunosorbent Assay, Infection, Diagnostic Assay

Journal: Retrovirology

Article Title: Increased expression of OX40 is associated with progressive disease in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/1742-4690-10-51

Figure Lengend Snippet: The expression of pro-inflammatory cytokines in peripheral blood mononuclear cells of HTLV-1 infected individuals

Article Snippet: Recombinant human OX40 ligand/TNFSF4 and recombinant human OX40/TNFRSF4/Fc Chimera were purchased from R&D Systems (Minneapolis, MN) and used for the standard curve in sOX40L and sOX40 ELISA, respectively.

Techniques: Expressing, Infection

Journal: Retrovirology

Article Title: Increased expression of OX40 is associated with progressive disease in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/1742-4690-10-51

Figure Lengend Snippet: Clinical and laboratory findings of HAM/TSP patients for whom paired CSF and plasma samples were tested for soluble OX40 (sOX40)

Article Snippet: Recombinant human OX40 ligand/TNFSF4 and recombinant human OX40/TNFRSF4/Fc Chimera were purchased from R&D Systems (Minneapolis, MN) and used for the standard curve in sOX40L and sOX40 ELISA, respectively.

Techniques:

Journal: Retrovirology

Article Title: Increased expression of OX40 is associated with progressive disease in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/1742-4690-10-51

Figure Lengend Snippet: Expression of OX40 in inflammatory mononuclear cells in spinal cord lesions of HAM/TSP patient with short disease duration and progressive symptoms. We studied autopsy specimens from 9 HAM/TSP patients by immunohistochemical staining. A. No OX40 positive cells are detected in the spinal cord lesion without active inflammation of a HAM/TSP patient with a long duration of illness. Magnification: ×40. B. Many infiltrating mononuclear cells are positively stained by anti-OX40 mAb in the spinal cord lesion with active inflammation of HAM/TSP patient with 2.5 years of illness. Magnification: ×40. C. There was reduced or no OX40L protein expression in spinal cord tissues of HAM/TSP patients. OX40L showed only low background staining and there was no OX40L positive staining on inflammatory mononuclear cells in the spinal cord lesions. Magnification: ×20. D. Positive control staining for OX40L positive CEM-OX40L cells. Magnification: ×20. Bar: 50 μm.

Article Snippet: Recombinant human OX40 ligand/TNFSF4 and recombinant human OX40/TNFRSF4/Fc Chimera were purchased from R&D Systems (Minneapolis, MN) and used for the standard curve in sOX40L and sOX40 ELISA, respectively.

Techniques: Expressing, Immunohistochemical staining, Staining, Positive Control

Journal: Retrovirology

Article Title: Increased expression of OX40 is associated with progressive disease in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis

doi: 10.1186/1742-4690-10-51

Figure Lengend Snippet: Anti-OX40 monoclonal antibody specifically eliminated naturally infected CD4 + T cells via antibody-dependent cell-mediated cytotoxicity (ADCC) in cultured PBMCs. Anti-OX40 mAb (clone B-7B5) reduces the percentage of Tax-positive cells, whereas the isotype control mAb (clone 2C2: anti-HIV-1 gp21, mouse IgG1) has no effect on Tax expression (1st, 2nd, and 3rd panels from left). Culture of PBMCs with anti-CD16/CD32 (Fc receptor) antibody to block Fc receptors abolishes Tax suppression by anti-OX40 mAb (4th panels from left). F(ab’) 2 fragment do not suppress Tax exppression (right panels).

Article Snippet: Recombinant human OX40 ligand/TNFSF4 and recombinant human OX40/TNFRSF4/Fc Chimera were purchased from R&D Systems (Minneapolis, MN) and used for the standard curve in sOX40L and sOX40 ELISA, respectively.

Techniques: Infection, Cell Culture, Expressing, Blocking Assay

Journal: Journal of Immunology Research

Article Title: Increased Expressions of OX40 and OX40 Ligand in Patients with Primary Immune Thrombocytopenia

doi: 10.1155/2019/6804806

Figure Lengend Snippet: Detection of antiplatelet autoantibodies in ITP patients. Twenty-two (41%) of 54 patients with primary ITP were positive for antiplatelet autoantibodies detected by ELISA. The peripheral platelet counts in patients with positive antiplatelet autoantibodies (ITP(+)) and negative antiplatelet autoantibodies (ITP(-)) and in HCs. ∗∗∗ p < 0.001; ∗∗ p < 0.01; ∗ p < 0.05.

Article Snippet: Plasma levels of sOX40L were analyzed by a Quantikine Human OX40 Ligand ELISA Kit (R&D Systems, Minneapolis, MN, USA) in accordance with the manufacturer's protocol.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Immunology Research

Article Title: Increased Expressions of OX40 and OX40 Ligand in Patients with Primary Immune Thrombocytopenia

doi: 10.1155/2019/6804806

Figure Lengend Snippet: The expression of OX40 on CD4 + T cells in ITP patients. Peripheral blood mononuclear cells (PBMCs) from 54 patients with ITP and 24 healthy controls (HCs) were isolated and stained with labeled antibodies and analyzed by flow cytometry, as described in . (a) The cells were gated initially on lymphocytes and then on CD4 + T cells. (b) The frequency of OX40 + CD4 + T cells among the total CD4 + T cells from HCs and ITP patients. (c) The relationship between the frequency of OX40 + CD4 + T cells and peripheral platelet counts in ITP patients with positive antiplatelet autoantibodies. (d) The relationship between the frequency of OX40 + CD4 + T cells and peripheral platelet counts in ITP patients with negative antiplatelet autoantibodies. ITP(+): ITP patients with positive antiplatelet autoantibodies; ITP(-): ITP patients with negative antiplatelet autoantibodies. ∗∗∗ p < 0.001; ∗∗ p < 0.01; ∗ p < 0.05.

Article Snippet: Plasma levels of sOX40L were analyzed by a Quantikine Human OX40 Ligand ELISA Kit (R&D Systems, Minneapolis, MN, USA) in accordance with the manufacturer's protocol.

Techniques: Expressing, Isolation, Staining, Labeling, Flow Cytometry

Journal: Journal of Immunology Research

Article Title: Increased Expressions of OX40 and OX40 Ligand in Patients with Primary Immune Thrombocytopenia

doi: 10.1155/2019/6804806

Figure Lengend Snippet: Plasma concentrations of soluble OX40L in patients. (a) Concentrations of plasma sOX40L in HCs and in ITP patients with negative and positive antiplatelet autoantibodies. (b) The relationship between the levels of plasma sOX40L and peripheral platelet counts in ITP patients with positive antiplatelet autoantibodies. (c) The relationship between the levels of plasma sOX40L and peripheral platelet counts in ITP patients with negative antiplatelet autoantibodies. ITP(+): ITP patients with positive antiplatelet autoantibodies; ITP(-): ITP patients with negative antiplatelet autoantibodies. ∗∗∗ p < 0.001; ∗∗ p < 0.01; ∗ p < 0.05.

Article Snippet: Plasma levels of sOX40L were analyzed by a Quantikine Human OX40 Ligand ELISA Kit (R&D Systems, Minneapolis, MN, USA) in accordance with the manufacturer's protocol.

Techniques: Clinical Proteomics

Journal: Journal of Immunology Research

Article Title: Increased Expressions of OX40 and OX40 Ligand in Patients with Primary Immune Thrombocytopenia

doi: 10.1155/2019/6804806

Figure Lengend Snippet: The expression of OX40 and OX40L mRNA in PBMCs of patients. (a) The expression levels of OX40 mRNA in PBMCs from 10 ITP patients with positive antiplatelet autoantibodies, 12 ITP patients with negative antiplatelet autoantibodies, and 12 HCs. (b) The expression levels of OX40L mRNA in PBMCs from 10 ITP patients with positive antiplatelet autoantibodies, 12 ITP patients with negative antiplatelet autoantibodies, and 12 HCs. ITP(+): ITP patients with positive antiplatelet autoantibodies; ITP(-): ITP patients with negative antiplatelet autoantibodies. ∗∗∗ p < 0.001; ∗∗ p < 0.01; ∗ p < 0.05.

Article Snippet: Plasma levels of sOX40L were analyzed by a Quantikine Human OX40 Ligand ELISA Kit (R&D Systems, Minneapolis, MN, USA) in accordance with the manufacturer's protocol.

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: Crucial role of OX40/OX40L signaling in a murine model of asthma

doi: 10.3892/mmr.2018.8453

Figure Lengend Snippet: Effects of OX40/OX40L signaling on recruitment of allergen-induced BALF eosinophils in response to OVA treatment. The eosinophil count in the BALF of OVA-treated mice was significantly increased. Furthermore, the number of eosinophils in BALF significantly increased in OX40L protein- and OX40 + T cell-treated mice sensitized and challenged with OVA compared with OVA-treated mice. Data are presented as percent of total cells ± SEM; n=6-8 mice/group; *P<0.05, **P<0.01. BALF, bronchoalveolar lavage fluid; OVA, ovalbumin; OX40L, OX40 ligand.

Article Snippet: Mouse recombinant OX40L protein was purchased from R&D Systems, Inc. (Minneapolis, MN, USA; catalog no. 1236-OX-025).

Techniques:

Journal: Molecular Medicine Reports

Article Title: Crucial role of OX40/OX40L signaling in a murine model of asthma

doi: 10.3892/mmr.2018.8453

Figure Lengend Snippet: Effects of OX40/OX40L signaling on cytokine secretion in BALF in response to OVA treatment. Expression levels of the cytokines IL-4, IL-6, IL-13, IL-17, TNF-α and IFN-γ were measured by ELISA in the BALF of PBS treated mice and in mice treated with OVA, OVA + OX40L protein, OVA + neutralizing anti-OX40L antibody, and OVA + OX40 + T cells. Data are shown as pg/ml ± SEM, n=6-8 mice/group; *P<0.05, **P<0.01. BALF, bronchoalveolar lavage fluid; IFN, interferon; IL, interleukin; OVA, ovalbumin; OX40L, OX40 ligand; TNF, tumor necrosis factor.

Article Snippet: Mouse recombinant OX40L protein was purchased from R&D Systems, Inc. (Minneapolis, MN, USA; catalog no. 1236-OX-025).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine Reports

Article Title: Crucial role of OX40/OX40L signaling in a murine model of asthma

doi: 10.3892/mmr.2018.8453

Figure Lengend Snippet: Effects of OX40/OX40L signaling effect on pNF-κB expression in BALF in response to OVA treatment. (A) Representative western blotting results from three independent experiments. (B) Ratios of pNF-κB to GAPDH protein bands of PBS treated mice and mice treated with OVA, OVA + OX40L protein, OVA + neutralizing anti-OX40L antibody, and OVA + OX40 + T cells. All values represent mean ± SEM; n=6-8 mice/group; *P<0.05. BALF, bronchoalveolar lavage fluid; NF, nuclear factor OVA, ovalbumin; OX40L, OX40 ligand.

Article Snippet: Mouse recombinant OX40L protein was purchased from R&D Systems, Inc. (Minneapolis, MN, USA; catalog no. 1236-OX-025).

Techniques: Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: Crucial role of OX40/OX40L signaling in a murine model of asthma

doi: 10.3892/mmr.2018.8453

Figure Lengend Snippet: Effects of OX40/OX40L signaling on T cell proliferation and polarization. (A and B) Flow cytometric analysis of OX40 + T cell polarization in vitro . Representative results from 3–4 tests of CD4 + /OX40 + T cells from control PBS-treated, OX40L protein-treated or OX40L protein + neutralizing anti-OX40L antibody-treated groups. Absorbance was detected using a microplate reader at a wavelength of 450 nm in cells treated with (C) different concentrations of OX40L protein (ng/ml) or 200 ng/ml OX40L protein + 200 ng/ml neutralizing anti-OX40L antibody; All values represent mean ± SEM; n=3; *P<0.05 vs. PBS group. (D) Proportion of OX40 + /OX40 + T cells from (A and B), respectively. All values are presented as the mean ± SEM; n=5; *P<0.05. NS, not significant; OX40L, OX40 ligand.

Article Snippet: Mouse recombinant OX40L protein was purchased from R&D Systems, Inc. (Minneapolis, MN, USA; catalog no. 1236-OX-025).

Techniques: In Vitro, Control

Journal: Molecular Medicine Reports

Article Title: Crucial role of OX40/OX40L signaling in a murine model of asthma

doi: 10.3892/mmr.2018.8453

Figure Lengend Snippet: Effects of OX40/OX40L signaling on protein expression of cleaved caspase-3 and cell apoptosis. (A) Protein extracts were obtained and subjected to western blotting for cleaved caspase-3 expression analysis. Representative results from three independent experiments are presented. (B) Ratios of cleaved caspase-3 to GAPDH protein bands of PBS-, OVA + OX40L protein-, and OVA + neutralizing anti-OX40L antibody-treated group. n=6-8; *P<0.05. (C) Representative results from 3–4 tests of Annexin V/propidium iodide-positive cells from either PBS-treated, OVA + OX40L-treated and OVA + neutralizing anti-OX40L antibody-treated mice. (D) The proportion of apoptotic cells from PBS, OVA + OX40L and OVA + neutralizing anti-OX40L antibody-treated mice was determined. n=6-8; All values are presented as the mean ± SEM; *P<0.05. OVA, ovalbumin; OX40L, OX40 ligand.

Article Snippet: Mouse recombinant OX40L protein was purchased from R&D Systems, Inc. (Minneapolis, MN, USA; catalog no. 1236-OX-025).

Techniques: Expressing, Western Blot

Journal: Molecules

Article Title: Combined OX40 Agonist and PD-1 Inhibitor Immunotherapy Improves the Efficacy of Vascular Targeted Photodynamic Therapy in a Urothelial Tumor Model

doi: 10.3390/molecules26123744

Figure Lengend Snippet: Figure 1. Combined treatment with OX40 agonist and PD1 inhibitor antibodies following VTP therapy suppresses primary tumor growth and improves survival. Percent survival of mice plotted by Kaplan-Meier methods (p < 0.0001).

Article Snippet: In the OX40 agonist subgroups, InVivoPlus antimouse OX40 CD134-Clone: OX-86 (500 mg/kg, Bioxcell, West Lebanon, NH, USA) was injected IP in a single dose on day 1 after VTP.

Techniques:

Journal: Molecules

Article Title: Combined OX40 Agonist and PD-1 Inhibitor Immunotherapy Improves the Efficacy of Vascular Targeted Photodynamic Therapy in a Urothelial Tumor Model

doi: 10.3390/molecules26123744

Figure Lengend Snippet: Figure 2. Adding PD-1 inhibitor and OX40 agonist to VTP therapy increases T cell infiltration and activation into MB-49 UTUC allografts at 7 days post-treatment and increases CD4+ and CD8+ T cell infiltration into MB-49 UTUC allografted tumors. (A) Heat map showing the mean frequency of each cell population as measured by flow cytometry in control and treated tumors. Each box represents the mean percentage of positive cells among the parent cell population, i.e., CD45+ among live cells, CD4+ or CD8+ cells among CD45+ cells, Foxp3+ Tregs or Teff among CD4+ cells, CD25+/Foxp3+

Article Snippet: In the OX40 agonist subgroups, InVivoPlus antimouse OX40 CD134-Clone: OX-86 (500 mg/kg, Bioxcell, West Lebanon, NH, USA) was injected IP in a single dose on day 1 after VTP.

Techniques: Activation Assay, Cytometry, Control

Journal: Molecules

Article Title: Combined OX40 Agonist and PD-1 Inhibitor Immunotherapy Improves the Efficacy of Vascular Targeted Photodynamic Therapy in a Urothelial Tumor Model

doi: 10.3390/molecules26123744

Figure Lengend Snippet: Figure 3. Adding PD-1 Inhibitor and OX40 agonist to VTP therapy increases T cell infiltration and activation compared to VTP alone, into MB-49 UTUC allografts at 7 days post-treatment. Representative flow cytometry plots showing the percentage of CD4+ and CD8+ cells among CD45+ cells (top row), Ki67+ in CD4+ (second row), Ki67+ in CD8+ (third row), Foxp3+ Tregs and Foxp3−Teff in CD4+ (fourth row), and GrzB+ in CD8+ T cells (bottom row) in control and treated tumors analyzed 7 days after treatment. A fluorescent minus one (FMO) sample was included for each marker as a negative control for gating.

Article Snippet: In the OX40 agonist subgroups, InVivoPlus antimouse OX40 CD134-Clone: OX-86 (500 mg/kg, Bioxcell, West Lebanon, NH, USA) was injected IP in a single dose on day 1 after VTP.

Techniques: Activation Assay, Flow Cytometry, Control, Marker, Negative Control