Journal: bioRxiv

Article Title: Adjuvant Discovery via a High Throughput Screen using Human Primary Mononuclear Cells

doi: 10.1101/2022.06.17.496630

Figure Lengend Snippet: A. Distribution of z scores, indicating induction of the cytokines (IFN, IL-10, and TNF). B. Distribution of z scores, representing the percentage of monocytes with activated HLA-DR and/or CD80/86 receptors. C. Distribution of z scores, representing the percentage of T-Cells with activated Ox40 receptors. D. Distribution of z scores, representing the percentage of B-Cells with activated HLA-DR and/or CD80/86 receptors. E. graphical representation of small molecule hit rate in varying number of donors. natural log of hit rate. F. Strong negative correlation between number of donors. Each dot represents one of the 2,296 compounds from the primary screen. The dashed line represents the donor threshold of a z score of 2 and representative z scores were chosen such that any compound above the threshold represents a hit in the screen.

Article Snippet: CD134 (OX40) Antibody, anti-human, APC, REAfinity TM (Clone- REA621) , Miltenyi Biotec , Order Number: 130-122-227.

Techniques:

Journal: bioRxiv

Article Title: Adjuvant Discovery via a High Throughput Screen using Human Primary Mononuclear Cells

doi: 10.1101/2022.06.17.496630

Figure Lengend Snippet: Key resource table

Article Snippet: CD134 (OX40) Antibody, anti-human, APC, REAfinity TM (Clone- REA621) , Miltenyi Biotec , Order Number: 130-122-227.

Techniques: Recombinant, Sterility, Injection, Modification, Staining, Saline, Enzyme-linked Immunosorbent Assay, Multiplex Assay, Software

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: Biomedical research (Tokyo, Japan)

Article Title: Blockade of CD80/CD86-CD28 co-stimulation augments the inhibitory function of peptide antigen-specific regulatory T cells.

doi: 10.2220/biomedres.45.115

Figure Lengend Snippet: Fig. 3 Expression of CD27, OX40, ICOS and 4-1BB on Foxp3 + CD4 + or Foxp3 − CD4 + DO 11.10 T cells. Expression of in- dicated co-stimulatory signal receptors on Foxp3 + CD4 + Treg cells and Foxp3 − CD4 + Tconv cells included in freshly isolat- ed DO 11.10, primed DO 11.10, and DO 11.10 iTS cells was examined by flow cytometry. The positive staining gate was set based on isotype-matched control antibody staining. Representative histograms from three independent experiments are presented.

Article Snippet: To assess the iTS population, the following mAbs were used: peridinin chlorophyll protein (PerCP)/Cyanine (Cy) 5.5-conjugated anti-mouse CD3 mAb (17A2, BioLegend), phycoerythrin (PE)/Cy7-conjugated anti-mouse CD4 mAb (GK1.5, BioLegend), fluorescein isothiocyanate (FITC)-conjugated anti-mouse TCR DO 11.10 mAb (KJ1-26, BioLegend), PE-conjugated anti-human CD2 mAb (RPA-2.10, Bio Legend), PE-conjugated anti-mouse CD28 mAb (37.51, Bio Legend), allophycocyanin (APC)-conjugated anti-mouse CD27 mAb (LG.3A10, BioLegend), APC-conjugated anti-mouse OX40 (CD134) mAb (OX-86; Proteintech, Rosemont, IL, USA), PE-conjugated anti-mouse ICOS (CD278) mAb (7E.17G9, Bio Legend), and APC-conjugated anti-mouse 4-1BB (CD137) mAb (17B5, Bio Legend).

Techniques: Expressing, Flow Cytometry, Staining, Control

Journal: Toxicology and applied pharmacology

Article Title: Decreased immune response in monkeys administered a human T-effector cell agonist (OX40) antibody.

doi: 10.1016/j.taap.2020.115285

Figure Lengend Snippet: Fig. 4. Decreased TDAR responses and increased soluble OX40 levels at 1 and 10 mg/kg/week of BMS 986178. A. TDAR responses. Data points represent individual monkey KLH-specific IgM and KLH-specific IgG (n = 3/sex/group) endpoint titer depicted in Log10 scale (y- axis). Horizontal bars represent group means. Day 1 samples were collected prior to dosing with KLH, while samples collected on Day 8, 15, 22, and 29 were 1, 2, 3, and 4 weeks post-KLH immunization, respectively. B. Serum soluble OX40 (sOX40) levels. Data points represent mean pg/mL of sOX40 detected in serum of vehicle and BMS 986178 dosed monkeys. Group means are displayed as horizontal black lines; n = 3/sex/group.

Article Snippet: Serum samples collected during the pretreatment period and prior to the second, third, and fifth weekly dose were analyzed for soluble OX40 using an electrochemiluminescence immunoassay method, with an OX40 specific antibody that does not compete with BMS-986178 (Bristol Myers Squibb) and a biotin-labeled polyclonal OX40 antibody (R&D Systems; Cat.# BAF3388) and Sulfo-tag-conjugated streptavidin (MesoScale Discovery Cat.# R32AD-1).

Techniques:

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: These sections were stained with either sheep anti-human OX40L polyclonal antibody (LS-B14343; LSBio, Seattle, WA, USA), rabbit anti-human OX40 polyclonal antibody (R&D Systems, Minneapolis, MN, USA) or isotype-matched control antibodies followed by ABC staining (Vector Lab, Burlingame, CA, USA).

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: These sections were stained with either sheep anti-human OX40L polyclonal antibody (LS-B14343; LSBio, Seattle, WA, USA), rabbit anti-human OX40 polyclonal antibody (R&D Systems, Minneapolis, MN, USA) or isotype-matched control antibodies followed by ABC staining (Vector Lab, Burlingame, CA, USA).

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: These sections were stained with either sheep anti-human OX40L polyclonal antibody (LS-B14343; LSBio, Seattle, WA, USA), rabbit anti-human OX40 polyclonal antibody (R&D Systems, Minneapolis, MN, USA) or isotype-matched control antibodies followed by ABC staining (Vector Lab, Burlingame, CA, USA).

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: These sections were stained with either sheep anti-human OX40L polyclonal antibody (LS-B14343; LSBio, Seattle, WA, USA), rabbit anti-human OX40 polyclonal antibody (R&D Systems, Minneapolis, MN, USA) or isotype-matched control antibodies followed by ABC staining (Vector Lab, Burlingame, CA, USA).

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: These sections were stained with either sheep anti-human OX40L polyclonal antibody (LS-B14343; LSBio, Seattle, WA, USA), rabbit anti-human OX40 polyclonal antibody (R&D Systems, Minneapolis, MN, USA) or isotype-matched control antibodies followed by ABC staining (Vector Lab, Burlingame, CA, USA).

Techniques: Injection, MANN-WHITNEY, Control