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: 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: eLife

Article Title: Single-cell glycomics analysis by CyTOF-Lec reveals glycan features defining cells differentially susceptible to HIV

doi: 10.7554/eLife.78870

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-Human CD134/OX40 (ACT35) (Mouse, Monoclonal) , Fluidigm , Cat# 3158012B , CyTOF (1:25).

Techniques: Recombinant, Plasmid Preparation, 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