anti ox40l antibody  (R&D Systems)

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: Western blotting analysis was conducted on the lysates of Hut78 cells. ( A ) Hut78 cells were treated with anti-OX40L neutralizing antibody (5 μg/mL) or isotype control for 0, 6, 12, or 24 h. Phosphorylation of AKT, ERK1/2, p38 MAPK, and JNK were measured. ( B ) Changes in phosphorylation of various signal transduction substances. * 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: Western Blot, Control, Phospho-proteomics, Transduction

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: Journal of Ophthalmic Inflammation and Infection

Article Title: OX40 ligand expression abrogates the immunosuppressive function of retinal pigment epithelium

doi: 10.1186/1869-5760-3-12

Figure Lengend Snippet: OX40L gene identification by PCR. PCR performed on purified plasmid with and without OX40L insert. Note the presence of OX40L in lanes 7 and 9. (Lane 6, low range DNA ladder; lane 7, eYFP-C1 + OX40L insert; lane 8, eYFP-C1 plasmid; lane 9, eYFP-N1 +OX40L insert; lane 10, eYFP-N1 plasmid).

Article Snippet: Biotinylated polyclonal goat anti-human OX40L antibody (Catalog # BAF1054) was obtained from R & D systems (Minneapolis, MN, USA).

Techniques: Purification, Plasmid Preparation

Journal: Journal of Ophthalmic Inflammation and Infection

Article Title: OX40 ligand expression abrogates the immunosuppressive function of retinal pigment epithelium

doi: 10.1186/1869-5760-3-12

Figure Lengend Snippet: Time trial after vector transfection. (a) Bar graph representing the expression pattern of the fluorescent protein (YFP) and OX40L after transfection of eYFP-C1-OX40L into RPE at five time points (time point 1 = 30 min, time point 2 = 1 h, time point 3 = 4 h, time point 4 = 6 h, and time point 5 = 24 h). Time-dependent increased expression is observed. (b) Flow cytometric analysis at 4 h after transfection of RPE cells with eYFP-C1-OX40L, with YFP expression (bold) in the left panel and OX40L expression (bold) in the right panel (the thinner lines represent RPE that was not transfected). (c) Bar graph expression pattern of YFP and OX40L after transfection of eYFP-N1-OX40L into RPE cells at the same time points. (d) Flow cytometric analysis at 4 h after transfection of RPE cells with eYFP-N1-OX40L.

Article Snippet: Biotinylated polyclonal goat anti-human OX40L antibody (Catalog # BAF1054) was obtained from R & D systems (Minneapolis, MN, USA).

Techniques: Plasmid Preparation, Transfection, Expressing

Journal: Journal of Ophthalmic Inflammation and Infection

Article Title: OX40 ligand expression abrogates the immunosuppressive function of retinal pigment epithelium

doi: 10.1186/1869-5760-3-12

Figure Lengend Snippet: RT-PCR demonstrated expression of OX40L in the RNA of ARPE cells stimulated by pro-inflammatory cytokines. Lanes 1 and 2 are two different DNA ladders; lanes 3 and 4 represent two different concentrations of the same RNA from stimulated RPE cells.

Article Snippet: Biotinylated polyclonal goat anti-human OX40L antibody (Catalog # BAF1054) was obtained from R & D systems (Minneapolis, MN, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Journal of Ophthalmic Inflammation and Infection

Article Title: OX40 ligand expression abrogates the immunosuppressive function of retinal pigment epithelium

doi: 10.1186/1869-5760-3-12

Figure Lengend Snippet: ARPE-19 and PBMC co-culture study results. (a) OX40L-C + PBMC abrogates the immunosuppressive effect of RPE alone. No difference was observed between OX40L (C- or N- vectors) or GITR ligand. An approximately 20% to 30% reversal of RPE-mediated immunosuppression is seen when stimulated pBMCs were co-cultured with RPE-OX40L vector. (b) A repeat experiment shows that concomitant transfection of both eYFP-C1-OX40L and eYFP-C1-GITRL vectors into RPE cells resulted into an additive reversal of immunosuppression of stimulated pBMCs, when comparing it to either RPE transfected with OX40L-C or GITRL alone. (pBMC unstim, unstimulated peripheral blood mononuclear cells; pBMC stim, stimulated pBMC with anti-CD3 and anti-CD28; RPE, retinal pigment epithelial cells without transfection; OX40L-C, RPE cells transfected with eYFP-C1-OX40L; OX40L-N, RPE cells transfected with eYFP-N1-OX40L; GITRL, RPE cells transfected with eYFP-C1-GITRL).

Article Snippet: Biotinylated polyclonal goat anti-human OX40L antibody (Catalog # BAF1054) was obtained from R & D systems (Minneapolis, MN, USA).

Techniques: Co-Culture Assay, Cell Culture, Plasmid Preparation, Transfection