Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: TL1A-DR3 plasma levels are predictive of HIV-1 disease control, and DR3 co-stimulation boosts HIV-1-specific T-cell responses

doi: 10.4049/jimmunol.2000933

Figure Lengend Snippet: sTL1A (A) and sDR3 (B) relative plasma levels (Z-score “communicome” values) in HIV-High (n=47) and HIV-Low (n=49) (Table S1). C-D) Gene expression (Table S1) of TL1A (C) and DR3 (D) in HIV-High (n=16) and HIV-Low (n=30). E-F) Unrelated validation cohorts for gene expression of TL1A (E) and DR3 (F) in dry pellet PBMC samples from seronegatives (SN, n=6) and HIV-infected individuals one year before and after initiation of treatment (untreated [n=11] and treated [n=5]) and in LTNP (n=23) (Table S1). G) Absolute DR3 quantifications in plasma (ELISA) in a confirmatory cohort including seronegatives (SN, n=8) and samples from time points one year before and after initiation of treatment (untreated [n=15] and treated [n=10]) and controllers (n=31), Table S1). The Mann-Whitney test was applied for group comparisons, and p-values < 0.05 were considered significant.

Article Snippet: Then, we co-stimulated the cells with 100 ng/ml of sTL1A (Recombinant Human TL1A/TNFSF15, R&D systems), 500 ng/ml of sDR3 (Recombinant Human DR3/TNFRSF25 Fc Chimera Protein, R&D systems), and /or 100 ng/ml of mAb anti-DR3 (Purified anti-human DR3 (TRAMP) JD3, Biolegend).

Techniques: Clinical Proteomics, Gene Expression, Biomarker Discovery, Infection, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: TL1A-DR3 plasma levels are predictive of HIV-1 disease control, and DR3 co-stimulation boosts HIV-1-specific T-cell responses

doi: 10.4049/jimmunol.2000933

Figure Lengend Snippet: A-B) sDR3 relative plasma levels (Z-score communicome) correlate with T-cell breadth (A) and magnitude (B) of HIV-High (n=46) and HIV-Low (n=41) individuals (Table S1). C) IFN-g-ELISPOT (percentage increase in the magnitude of T-cell) after HIV-specific peptide pool stimulation in the presence of recombinant TL1A and DR3 and specific anti-DR3 mAb in PBMCs from untreated PLWH (Table S1). D) Percentage increase in the magnitude of CEF-specific responses upon co-stimulation with anti-DR3 mAb. E) Increase in magnitude of DNA.HTI-specific T cell responses in isolated splenocytes from HTI-vaccinated mice after stimulation with the mouse DR3 specific antibody 4C12 (agonist DR3). The Spearman rank test was used for correlation analysis and the Wilcoxon signed rank test for group comparisons. p-values <0.05 were considered statistically significant.

Article Snippet: Then, we co-stimulated the cells with 100 ng/ml of sTL1A (Recombinant Human TL1A/TNFSF15, R&D systems), 500 ng/ml of sDR3 (Recombinant Human DR3/TNFRSF25 Fc Chimera Protein, R&D systems), and /or 100 ng/ml of mAb anti-DR3 (Purified anti-human DR3 (TRAMP) JD3, Biolegend).

Techniques: Clinical Proteomics, Enzyme-linked Immunospot, Recombinant, Isolation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Galectin-9 binds to DR3. (a) Immunoprecipitation of recombinant human or mouse Galectin-9 with human DR3.Fc or 4-1BB.Fc. Control, human Galectin-3. (b) Immunoprecipitation of recombinant human or mouse Galectin-9 with mouse DR3.Fc. Observed molecular masses were h/m4-1BB.Fc or DR3.Fc – 50–58 kDa, m/hGalectin-9 – 36 kDa. Data in (a–b) are representative of at least three different experiments each. (c–f) Binding response of increasing concentrations (0.48–2000 nM) of hGalectin-9 (c, d) or mGalectin-9 (e, f) to immobilized hDR3.Fc (c, e) or mDR3.Fc (d, f), measured by SPR. Values represent average of three independent measurements. The response shown is reference-subtracted (unrelated Fc protein). (g–h) Human or mouse Galectin-9 or TL1A were coated onto ELISA plates and binding of WT or de-glycosylated DR3.Fc was detected using biotin anti-DR3 and streptavidin HRP. Competition was assessed with DR3.Fc first incubated with either TL1A or Galectin-9. Data are means ± s.e.m binding from triplicate cultures and representative of three different experiments.

Article Snippet: Fc fusion proteins, TL1A, and Galectin-9 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Immunoprecipitation, Recombinant, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Defective DR3 activity in T cells from Galectin-9−/− mice. (a) Naïve CD4+ T cells from WT or Galectin-9−/− mice were activated in vitro with anti-CD3 and anti-CD28 in the presence of control IgG and agonist TL1A.Ig. IL-2 and IFN-γ production were assessed after 48 h. (b) CD4+ T cells from WT and Galectin-9−/− mice were pre-activated with anti-CD3 and anti-CD28, and then after 48 h were restimulated with anti-CD3 in the presence of control IgG or agonist anti-DR3. IL-2 and IFN-γ production were assessed 48 h later. (c) CD4+ T cells from WT (solid line) and Galectin-9−/− (dotted line) mice were pre-activated as in (b) and stained for DR3 expression (pre-sort, left). Isotype control staining, shaded. Sorted cells were then restimulated with anti-CD3 in the presence of control IgG or agonist TL1A.Ig. IL-2 production was assessed at 24 h. All data are means ± s.e.m from triplicate cultures and representative of three different experiments. *p<0.05

Article Snippet: Fc fusion proteins, TL1A, and Galectin-9 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Activity Assay, In Vitro, Staining, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Treg from Galectin-9−/− mice are refractory to stimulation through DR3. (a) CD4+Foxp3+ Treg cells from WT mice were stained for surface DR3 and intracellular Galectin-9. Shaded, isotype control. Dotted line, Treg from Galectin-9−/− mice. (b) Immunoprecipitation with TL1A.Ig or control Fc was performed on lysates from WT Treg cells. Immunoblotting was carried out with anti-DR3 (top) or anti-Galectin-9 (bottom). MW indicated. Data are representative of three experiments. (c) Treg cells from WT or Galectin-9−/− mice were stimulated with anti-CD3 and IL-2 in the presence of control IgG or agonist TL1A.Ig. Proliferation was assessed at 72 h. Data are means ± s.e.m from triplicate cultures and representative of three experiments. *Significance TL1A.Ig WT vs. Galectin-9−/−. (d) WT (solid line) and Galectin-9−/− (dotted line) Treg cells were stimulated with anti-CD3 and IL-2 in the presence of control IgG or agonist anti-DR3. After 48 h, cells were stained for intracellular IDO and IL-10. MFI indicated. Isotype controls, shaded. Data are representative of three different experiments. *p<0.05

Article Snippet: Fc fusion proteins, TL1A, and Galectin-9 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Staining, Immunoprecipitation, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Galectin-9 is required for suppression of EAE by anti-DR3. WT or Galectin-9−/− mice were immunized with MOG35-55 peptide and injected with either control IgG or agonist anti-DR3. (a) Percent Foxp3+CD4+ T cells in peripheral blood 5 days after the last injection of anti-DR3. (b) EAE clinical scores over time. (c) Frequencies of IL-17A+ CD4 T cells (left) and TNF+ CD4 T cells (middle and right) in draining lymph nodes on days 17 and 30, respectively. (d) Proportion of gated CD4+ cells expressing IL-10 and Foxp3 (left) and total numbers of Foxp3+IL-10+ CD4 T cells (right) in brains of anti-DR3-treated animals at day 17. (e) Proportion of gated CD4+ cells expressing IL-17 or IFN-γ (left) and total numbers of IL-17+ CD4 T cells (right) in brains of anti-DR3-treated animals at day 17. All data are either representative or means ± sem from five mice per group. Similar results in three different experiments. *p<0.05

Article Snippet: Fc fusion proteins, TL1A, and Galectin-9 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Injection, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Galectin-9 is required for suppression of allergic asthma by anti-DR3. WT and Galectin-9−/− mice were immunized with OVA to induce lung inflammation, and injected with IgG or agonist anti-DR3. (a) Representative H&E staining of lung sections (left), and mean inflammation score (right). (b) Eosinophil numbers and IL-5 expression in BAL. (c) Proportion of CD4+Foxp3+ T cells in BAL (left) and draining lymph nodes (right). All results are means ± s.e.m from five mice per group, and representative of three independent experiments. *p<0.05

Article Snippet: Fc fusion proteins, TL1A, and Galectin-9 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Injection, Staining, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Galectin-9 binds to DR3. (a) Immunoprecipitation of recombinant human or mouse Galectin-9 with human DR3.Fc or 4-1BB.Fc. Control, human Galectin-3. (b) Immunoprecipitation of recombinant human or mouse Galectin-9 with mouse DR3.Fc. Observed molecular masses were h/m4-1BB.Fc or DR3.Fc – 50–58 kDa, m/hGalectin-9 – 36 kDa. Data in (a–b) are representative of at least three different experiments each. (c–f) Binding response of increasing concentrations (0.48–2000 nM) of hGalectin-9 (c, d) or mGalectin-9 (e, f) to immobilized hDR3.Fc (c, e) or mDR3.Fc (d, f), measured by SPR. Values represent average of three independent measurements. The response shown is reference-subtracted (unrelated Fc protein). (g–h) Human or mouse Galectin-9 or TL1A were coated onto ELISA plates and binding of WT or de-glycosylated DR3.Fc was detected using biotin anti-DR3 and streptavidin HRP. Competition was assessed with DR3.Fc first incubated with either TL1A or Galectin-9. Data are means ± s.e.m binding from triplicate cultures and representative of three different experiments.

Article Snippet: 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Immunoprecipitation, Recombinant, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Defective DR3 activity in T cells from Galectin-9−/− mice. (a) Naïve CD4+ T cells from WT or Galectin-9−/− mice were activated in vitro with anti-CD3 and anti-CD28 in the presence of control IgG and agonist TL1A.Ig. IL-2 and IFN-γ production were assessed after 48 h. (b) CD4+ T cells from WT and Galectin-9−/− mice were pre-activated with anti-CD3 and anti-CD28, and then after 48 h were restimulated with anti-CD3 in the presence of control IgG or agonist anti-DR3. IL-2 and IFN-γ production were assessed 48 h later. (c) CD4+ T cells from WT (solid line) and Galectin-9−/− (dotted line) mice were pre-activated as in (b) and stained for DR3 expression (pre-sort, left). Isotype control staining, shaded. Sorted cells were then restimulated with anti-CD3 in the presence of control IgG or agonist TL1A.Ig. IL-2 production was assessed at 24 h. All data are means ± s.e.m from triplicate cultures and representative of three different experiments. *p<0.05

Article Snippet: 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Activity Assay, In Vitro, Staining, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Treg from Galectin-9−/− mice are refractory to stimulation through DR3. (a) CD4+Foxp3+ Treg cells from WT mice were stained for surface DR3 and intracellular Galectin-9. Shaded, isotype control. Dotted line, Treg from Galectin-9−/− mice. (b) Immunoprecipitation with TL1A.Ig or control Fc was performed on lysates from WT Treg cells. Immunoblotting was carried out with anti-DR3 (top) or anti-Galectin-9 (bottom). MW indicated. Data are representative of three experiments. (c) Treg cells from WT or Galectin-9−/− mice were stimulated with anti-CD3 and IL-2 in the presence of control IgG or agonist TL1A.Ig. Proliferation was assessed at 72 h. Data are means ± s.e.m from triplicate cultures and representative of three experiments. *Significance TL1A.Ig WT vs. Galectin-9−/−. (d) WT (solid line) and Galectin-9−/− (dotted line) Treg cells were stimulated with anti-CD3 and IL-2 in the presence of control IgG or agonist anti-DR3. After 48 h, cells were stained for intracellular IDO and IL-10. MFI indicated. Isotype controls, shaded. Data are representative of three different experiments. *p<0.05

Article Snippet: 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Staining, Immunoprecipitation, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Galectin-9 is required for suppression of EAE by anti-DR3. WT or Galectin-9−/− mice were immunized with MOG35-55 peptide and injected with either control IgG or agonist anti-DR3. (a) Percent Foxp3+CD4+ T cells in peripheral blood 5 days after the last injection of anti-DR3. (b) EAE clinical scores over time. (c) Frequencies of IL-17A+ CD4 T cells (left) and TNF+ CD4 T cells (middle and right) in draining lymph nodes on days 17 and 30, respectively. (d) Proportion of gated CD4+ cells expressing IL-10 and Foxp3 (left) and total numbers of Foxp3+IL-10+ CD4 T cells (right) in brains of anti-DR3-treated animals at day 17. (e) Proportion of gated CD4+ cells expressing IL-17 or IFN-γ (left) and total numbers of IL-17+ CD4 T cells (right) in brains of anti-DR3-treated animals at day 17. All data are either representative or means ± sem from five mice per group. Similar results in three different experiments. *p<0.05

Article Snippet: 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Injection, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Treg-mediated Suppression of Inflammation Induced by DR3 Signaling is Dependent on Galectin-9

doi: 10.4049/jimmunol.1700575

Figure Lengend Snippet: Galectin-9 is required for suppression of allergic asthma by anti-DR3. WT and Galectin-9−/− mice were immunized with OVA to induce lung inflammation, and injected with IgG or agonist anti-DR3. (a) Representative H&E staining of lung sections (left), and mean inflammation score (right). (b) Eosinophil numbers and IL-5 expression in BAL. (c) Proportion of CD4+Foxp3+ T cells in BAL (left) and draining lymph nodes (right). All results are means ± s.e.m from five mice per group, and representative of three independent experiments. *p<0.05

Article Snippet: 4-1BB.Fc was produced as described previously ( 7 ) and mouse and human DR3.Fc were from either Enzo Life Sciences or R&D Systems.

Techniques: Injection, Staining, Expressing