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    Structured Review

    Thermo Fisher anti il 10 ab
    <t>IL-10</t> inhibits FcεRI-dependent TNF-α expression by blood mDCs

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    Images

    1) Product Images from "Interferons Modulate Fc?RI-dependent Production of Autoregulatory IL-10 by Circulating Human Monocytoid Dendritic Cells"

    Article Title: Interferons Modulate Fc?RI-dependent Production of Autoregulatory IL-10 by Circulating Human Monocytoid Dendritic Cells

    Journal:

    doi: 10.1016/j.jaci.2008.09.013

    IL-10 inhibits FcεRI-dependent TNF-α expression by blood mDCs
    Figure Legend Snippet: IL-10 inhibits FcεRI-dependent TNF-α expression by blood mDCs

    Techniques Used: Expressing

    TLR9-induced IFN-α from pDCs enhances FcεRI-dependent IL-10 secretion by mDCs
    Figure Legend Snippet: TLR9-induced IFN-α from pDCs enhances FcεRI-dependent IL-10 secretion by mDCs

    Techniques Used:

    Kinetics for FcεRI-dependent TNF-α and IL-10 secretion by blood mDCs
    Figure Legend Snippet: Kinetics for FcεRI-dependent TNF-α and IL-10 secretion by blood mDCs

    Techniques Used:

    Inhibitory effects of IL-10 on FcεRI-dependent responses in mDCs are not from a down-regulation in FcεRIα and co-stimulatory molecule expression
    Figure Legend Snippet: Inhibitory effects of IL-10 on FcεRI-dependent responses in mDCs are not from a down-regulation in FcεRIα and co-stimulatory molecule expression

    Techniques Used: Expressing

    Interferons modulate FcεRI-dependent IL-10 secretion by mDCs
    Figure Legend Snippet: Interferons modulate FcεRI-dependent IL-10 secretion by mDCs

    Techniques Used:

    2) Product Images from "Programmed Death-Ligand 1 Expression Potentiates the Immune Modulatory Function Of Myeloid-Derived Suppressor Cells in Systemic Lupus Erythematosus"

    Article Title: Programmed Death-Ligand 1 Expression Potentiates the Immune Modulatory Function Of Myeloid-Derived Suppressor Cells in Systemic Lupus Erythematosus

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2021.606024

    In vivo treatment with PD-L1 positive MDSCs shows better therapeutic effects in lupus prone Roquin san/san mice. (A) 13-week-old Roquin san/san mice (n=5 in each group) were treated intravenously with 1 × 10 6 PD-L1 positive and/or negative MDSCs from C57BL/6 mice or vehicle (saline) once a week for 2 weeks. One week after the last treatment, Roquin san/san mice was sacrificed. (B) Serum levels of IgG2a and anti-dsDNA IgG in Roquin san/san mice treated with PD-L1 positive or negative MDSCs or saline, as determined by ELISA. (C) Representative images of hematoxylin and eosin (H E) and PAS staining of the kidney sections are shown in the left panel. Tubular injury score is shown in the right panel. (D) Flow cytometric analysis shows the populations of CD4+IFN-γ T cells (Th1 cells), CD4+IL-4+ T cells (Th2 cells), CD4+IL-17+ T cells (Th17 cells), CD4+CD25+FOXP3+ cells (Treg cells) in spleen tissues from Roquin san/san mice. Data are expressed as the mean ± SEM. Data are representative of 2 independent experiments. Relative bar charts are shown. (E) Flow cytometric analysis shows the populations of B220-CD138+ cells (plasma cells) and CD19+CD1d+CD5+IL-10+ cells (IL-10 producing B10 cells) in spleen tissues from Roquin san/san mice. Relative bar charts are shown. (* P
    Figure Legend Snippet: In vivo treatment with PD-L1 positive MDSCs shows better therapeutic effects in lupus prone Roquin san/san mice. (A) 13-week-old Roquin san/san mice (n=5 in each group) were treated intravenously with 1 × 10 6 PD-L1 positive and/or negative MDSCs from C57BL/6 mice or vehicle (saline) once a week for 2 weeks. One week after the last treatment, Roquin san/san mice was sacrificed. (B) Serum levels of IgG2a and anti-dsDNA IgG in Roquin san/san mice treated with PD-L1 positive or negative MDSCs or saline, as determined by ELISA. (C) Representative images of hematoxylin and eosin (H E) and PAS staining of the kidney sections are shown in the left panel. Tubular injury score is shown in the right panel. (D) Flow cytometric analysis shows the populations of CD4+IFN-γ T cells (Th1 cells), CD4+IL-4+ T cells (Th2 cells), CD4+IL-17+ T cells (Th17 cells), CD4+CD25+FOXP3+ cells (Treg cells) in spleen tissues from Roquin san/san mice. Data are expressed as the mean ± SEM. Data are representative of 2 independent experiments. Relative bar charts are shown. (E) Flow cytometric analysis shows the populations of B220-CD138+ cells (plasma cells) and CD19+CD1d+CD5+IL-10+ cells (IL-10 producing B10 cells) in spleen tissues from Roquin san/san mice. Relative bar charts are shown. (* P

    Techniques Used: In Vivo, Mouse Assay, Enzyme-linked Immunosorbent Assay, Staining

    PD-L1 expressing MDSCs from control mice shows more powerful immune regulatory activity in vitro . (A) PD-L1 positive MDSCs from lupus prone MRL/ lpr mice and MLR/MpJ (control mice) were isolated. The mRNA levels of pro or anti-inflammatory molecules were determined. (B) CD4+ T cells (5 × 10 5 ) from MLR/ lpr mice were labeled with Cell trace violet, and cultured with/without PD-L1 positive or negative MDSCs (1:1, 5:1 ratio). The percentage of proliferating CD4+ cells were analyzed. (C) CD4+ T cells from MLR/ lpr mice were cultured with PD-L1 positive MDSCs from MRL/ lpr mice and MLR/MpJ (1:1 ratio) in the presence of anti-CD3 (0.5 μ/ml). After 3 days, cells were analyzed for Th17, Th1 and Treg cells by flow cytometry. The percentage of each cell population are shown in the right panel. The concentrations of IL-17, IFN- γ and TGF-β in supernatant were measured by ELISA. (D) Splenocytes from MLR/ lpr mice were cultured with PD-L1 positive MDSCs (1:1 ratio) in the presence of LPS (100 ng/ml). After 3 days, cells were analyzed for IL-10 producing B10 cells and plasma cells (CD19-CD138+) by flow cytometry. The percentage of each cell population are shown in the right panel. The concentrations of IL-10 and IL-6 in supernatant were measured. Data are expressed as the mean ± SEM. Data are representative of three independent experiments (* P
    Figure Legend Snippet: PD-L1 expressing MDSCs from control mice shows more powerful immune regulatory activity in vitro . (A) PD-L1 positive MDSCs from lupus prone MRL/ lpr mice and MLR/MpJ (control mice) were isolated. The mRNA levels of pro or anti-inflammatory molecules were determined. (B) CD4+ T cells (5 × 10 5 ) from MLR/ lpr mice were labeled with Cell trace violet, and cultured with/without PD-L1 positive or negative MDSCs (1:1, 5:1 ratio). The percentage of proliferating CD4+ cells were analyzed. (C) CD4+ T cells from MLR/ lpr mice were cultured with PD-L1 positive MDSCs from MRL/ lpr mice and MLR/MpJ (1:1 ratio) in the presence of anti-CD3 (0.5 μ/ml). After 3 days, cells were analyzed for Th17, Th1 and Treg cells by flow cytometry. The percentage of each cell population are shown in the right panel. The concentrations of IL-17, IFN- γ and TGF-β in supernatant were measured by ELISA. (D) Splenocytes from MLR/ lpr mice were cultured with PD-L1 positive MDSCs (1:1 ratio) in the presence of LPS (100 ng/ml). After 3 days, cells were analyzed for IL-10 producing B10 cells and plasma cells (CD19-CD138+) by flow cytometry. The percentage of each cell population are shown in the right panel. The concentrations of IL-10 and IL-6 in supernatant were measured. Data are expressed as the mean ± SEM. Data are representative of three independent experiments (* P

    Techniques Used: Expressing, Mouse Assay, Activity Assay, In Vitro, Isolation, Labeling, Cell Culture, Flow Cytometry, Enzyme-linked Immunosorbent Assay

    Cell therapy with PD-L1 positive MDSCs profoundly decreases pathogenic double negative T cells (DN T cells) in spleens of MRL/ lpr mice. (A) Flow cytometric analysis shows the populations of DN T cells (CD4-CD8-CD3+ T cells), CD4+ T cells, CD8+ T cells, IL2+CD4+ T cells, CD4+IL-10+ T cells, CD4+IFN-γ T cells (Th1 cells), CD4+IL-17+ T cells (Th17 cells), CD4+IL-4+ T cells (Th2 cells), CD4+CD25+FOXP3+ cells (Treg cells), CD4+CXCR5+PD-1+ T cells (Tfh cells) and CD4+CXCR5+PD-1+ Foxp3+(TFR cells) in spleen tissues from MRL/ lpr mice treated with PD-L1 positive and/or negative MDSCs obtained from MRL/MpJ mice. Relative bar charts are shown. (B) Flow cytometric analysis shows the populations of CD19+GL7+ cells (GC B cells), CD19-CD138+ cells (plasma cells) and CD19+CD1d+CD5+IL-10+ cells (IL-10 producing B10 cells) in spleen tissues from MRL/ lpr mice. Relative bar charts are shown. Data are expressed as the mean ± SEM. Data are representative of three independent experiments [* P
    Figure Legend Snippet: Cell therapy with PD-L1 positive MDSCs profoundly decreases pathogenic double negative T cells (DN T cells) in spleens of MRL/ lpr mice. (A) Flow cytometric analysis shows the populations of DN T cells (CD4-CD8-CD3+ T cells), CD4+ T cells, CD8+ T cells, IL2+CD4+ T cells, CD4+IL-10+ T cells, CD4+IFN-γ T cells (Th1 cells), CD4+IL-17+ T cells (Th17 cells), CD4+IL-4+ T cells (Th2 cells), CD4+CD25+FOXP3+ cells (Treg cells), CD4+CXCR5+PD-1+ T cells (Tfh cells) and CD4+CXCR5+PD-1+ Foxp3+(TFR cells) in spleen tissues from MRL/ lpr mice treated with PD-L1 positive and/or negative MDSCs obtained from MRL/MpJ mice. Relative bar charts are shown. (B) Flow cytometric analysis shows the populations of CD19+GL7+ cells (GC B cells), CD19-CD138+ cells (plasma cells) and CD19+CD1d+CD5+IL-10+ cells (IL-10 producing B10 cells) in spleen tissues from MRL/ lpr mice. Relative bar charts are shown. Data are expressed as the mean ± SEM. Data are representative of three independent experiments [* P

    Techniques Used: Mouse Assay

    Characterization of MDSCs from SLE patients. (A) MDSCs subpopulations of human peripheral bloods was determined by flow cytometry in SLE patients and healthy controls (Total MDSC: HLA-DR-Lin-CD33+CD11b+ cells, M-MDSC: CD14+ HLA-DR- cells, PMN-MDSC: CD14-CD15+CD11b+ cells). Gating strategy of human MDSCs and representative flow cytometric plot is shown. (B) Relative bar graph indicates the frequency of MDSC subpopulation in healthy controls and SLE patients. (C) The expression of PD-L1, arginase-1, and IL-10 was determined by flow cytometry. The percentage of PD-L1, Arginase-1, and IL-10 expressing MDSCs is shown. (D) Representative confocal microscopy analysis for MDSC like cells in kidney of patients with lupus nephritis and healthy controls. Cells were stained with fluorescence-tagged antibodies to identify DAPI, CD11b (FITC) and CD33 (PE). CD11b+CD33+ cells, MDSC like cells, are located in kidney tissue sections, especially in patients with lupus nephritis (x100(left), x400(right), original magnification). (* P
    Figure Legend Snippet: Characterization of MDSCs from SLE patients. (A) MDSCs subpopulations of human peripheral bloods was determined by flow cytometry in SLE patients and healthy controls (Total MDSC: HLA-DR-Lin-CD33+CD11b+ cells, M-MDSC: CD14+ HLA-DR- cells, PMN-MDSC: CD14-CD15+CD11b+ cells). Gating strategy of human MDSCs and representative flow cytometric plot is shown. (B) Relative bar graph indicates the frequency of MDSC subpopulation in healthy controls and SLE patients. (C) The expression of PD-L1, arginase-1, and IL-10 was determined by flow cytometry. The percentage of PD-L1, Arginase-1, and IL-10 expressing MDSCs is shown. (D) Representative confocal microscopy analysis for MDSC like cells in kidney of patients with lupus nephritis and healthy controls. Cells were stained with fluorescence-tagged antibodies to identify DAPI, CD11b (FITC) and CD33 (PE). CD11b+CD33+ cells, MDSC like cells, are located in kidney tissue sections, especially in patients with lupus nephritis (x100(left), x400(right), original magnification). (* P

    Techniques Used: Flow Cytometry, Expressing, Confocal Microscopy, Staining, Fluorescence

    PD-L1 expressing MDSCs have stronger in vitro immunoregulatory activity in mice. (A) MDSCs were obtained from C57BL/6 mice as described in Materials and Methods section. MDSCs sorted into PD-L1+ and PD-L1− sub-populations using a cell sorter. (A) The mRNA levels of IL-10, arginase-1, iNOS and TGF-β in PD-L1 positive or negative MDSCs were determined using real-time PCR. (B) CD4+T cells (5 × 10 5 ) were cultured with PD-L1 positive or negative MDSCs in the presence of anti-CD3 (0.5 μ/ml). After 3 days, cells were analyzed for Treg (CD4+CD25+Foxp3+) and Th17 (CD4+IL-17+) cells. The representative flow cytometric plot is shown. The percentage of each cell population among CD4+ T cells are shown in the right upper panel. The concentrations of IL-17 and IFN-γ in culture supernatant were measured by ELISA. (C) Splenocytes (5 × 10 5 ) were cultured with PD-L1 positive or negative MDSCs in the presence of LPS (100 ng/ml). After 3 days, cells were analyzed for IL-10 producing B10 cells (CD19+CD1d+CD5+ cells) by flow cytometry. The percentage of each cell population are shown in the right panel. The concentrations of IL-10 in supernatant were measured by ELISA. Data are expressed as the mean ± SEM. Data are representative of three independent experiments (* P
    Figure Legend Snippet: PD-L1 expressing MDSCs have stronger in vitro immunoregulatory activity in mice. (A) MDSCs were obtained from C57BL/6 mice as described in Materials and Methods section. MDSCs sorted into PD-L1+ and PD-L1− sub-populations using a cell sorter. (A) The mRNA levels of IL-10, arginase-1, iNOS and TGF-β in PD-L1 positive or negative MDSCs were determined using real-time PCR. (B) CD4+T cells (5 × 10 5 ) were cultured with PD-L1 positive or negative MDSCs in the presence of anti-CD3 (0.5 μ/ml). After 3 days, cells were analyzed for Treg (CD4+CD25+Foxp3+) and Th17 (CD4+IL-17+) cells. The representative flow cytometric plot is shown. The percentage of each cell population among CD4+ T cells are shown in the right upper panel. The concentrations of IL-17 and IFN-γ in culture supernatant were measured by ELISA. (C) Splenocytes (5 × 10 5 ) were cultured with PD-L1 positive or negative MDSCs in the presence of LPS (100 ng/ml). After 3 days, cells were analyzed for IL-10 producing B10 cells (CD19+CD1d+CD5+ cells) by flow cytometry. The percentage of each cell population are shown in the right panel. The concentrations of IL-10 in supernatant were measured by ELISA. Data are expressed as the mean ± SEM. Data are representative of three independent experiments (* P

    Techniques Used: Expressing, In Vitro, Activity Assay, Mouse Assay, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry

    3) Product Images from "Prostaglandin E2 and SOCS1 have a role in intestinal immune tolerance"

    Article Title: Prostaglandin E2 and SOCS1 have a role in intestinal immune tolerance

    Journal: Nature Communications

    doi: 10.1038/ncomms1181

    IFNγ renders Socs1 −/− BMDCs resistant to PGE2 but not to IL-10. ( a – e , g , h ) Socs1 +/+ (black bars) and Socs1 −/− (grey bars) BMDCs (5×10 5 cells per ml) induced from the bone marrow cells of Rag2 −/− and Socs1 −/− Rag2 −/− mice were stimulated for 12 h with indicated reagents, and the levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. Error bars represent +s.d. Cells were stimulated in triplicate wells for each condition and s.d. was calculated from the values determined by ELISA. ( a ) Socs1 +/+ and Socs1 −/− BMDCs were stimulated with LPS (10 ng ml −1 ) and murine IFNγ (10 ng ml −1 ) in the presence of graded concentrations of recombinant murine IL-10. ( b ) Socs1 +/+ and Socs1 −/− BMDCs were co-cultured with 2.5×10 5 cells per ml of CD4 + CD25 high T cells in the presence of soluble anti-CD3 Ab (1 μg ml −1 ). Cells were stimulated with LPS+IFNγ (10 ng ml −1 each) with or without anti-IL-10-neutralizing Ab (αIL-10 Ab, 10 μg ml −1 ). ( c ) Socs1 +/+ BMDCs were stimualted with LPS (10 ng ml −1 ) in the presence or absence of graded concentrations of CME and αIL-10 Ab (10 μg ml −1 ). ( d ) Socs1 +/+ and Socs1 −/− BMDCs were stimulated with LPS+IFNγ (10 ng ml −1 each) in the presence of graded concentrations of CME (v/v, volume of CME/volume of solution (media+CME)). ** P
    Figure Legend Snippet: IFNγ renders Socs1 −/− BMDCs resistant to PGE2 but not to IL-10. ( a – e , g , h ) Socs1 +/+ (black bars) and Socs1 −/− (grey bars) BMDCs (5×10 5 cells per ml) induced from the bone marrow cells of Rag2 −/− and Socs1 −/− Rag2 −/− mice were stimulated for 12 h with indicated reagents, and the levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. Error bars represent +s.d. Cells were stimulated in triplicate wells for each condition and s.d. was calculated from the values determined by ELISA. ( a ) Socs1 +/+ and Socs1 −/− BMDCs were stimulated with LPS (10 ng ml −1 ) and murine IFNγ (10 ng ml −1 ) in the presence of graded concentrations of recombinant murine IL-10. ( b ) Socs1 +/+ and Socs1 −/− BMDCs were co-cultured with 2.5×10 5 cells per ml of CD4 + CD25 high T cells in the presence of soluble anti-CD3 Ab (1 μg ml −1 ). Cells were stimulated with LPS+IFNγ (10 ng ml −1 each) with or without anti-IL-10-neutralizing Ab (αIL-10 Ab, 10 μg ml −1 ). ( c ) Socs1 +/+ BMDCs were stimualted with LPS (10 ng ml −1 ) in the presence or absence of graded concentrations of CME and αIL-10 Ab (10 μg ml −1 ). ( d ) Socs1 +/+ and Socs1 −/− BMDCs were stimulated with LPS+IFNγ (10 ng ml −1 each) in the presence of graded concentrations of CME (v/v, volume of CME/volume of solution (media+CME)). ** P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Cell Culture

    IFNγ antagonizes cAMP but not Treg-derived IL-10 in Socs1 −/− BMDCs. ( a ) Socs1 +/+ (5×10 5 cells per ml; black bars) and Socs1 −/− BMDCs (5×10 5 cells per ml; grey bars) were stimulated with LPS+IFNγ (10 ng ml −1 each) for 12 h in the presence or absence of graded concentrations of membrane-permeable cAMP analogue, 8-Br-cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. ( b ) Socs1 +/+ (5×10 5 cells per ml; left half of each panel) and Socs1 −/− BMDCs (5×10 5 cells per ml; right half of each panel) were stimulated with LPS (10 ng ml −1 ) for 12 h with (white bars) or without (black bars) 100 μM of 8-Br-cAMP with graded concentrations of murine recombinant IFNγ. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. IFNγ dose-dependently antagonized cAMP-mediated immunosuppression, which was enhanced by SOCS1 deficiency. ( c ) Stat1 +/+ (5×10 5 cells per ml; white bars) and Stat1 −/− (5×10 5 cells per ml; grey bars) BMDCs were stimulated with LPS (10 ng ml −1 ) for 24 h in the absence or presence of PGE2 and graded concentrations of murine recombinant IFNγ. The level of TNFα in the culture supernatant was measured by ELISA. ( d ) Socs1 +/+ and Socs1 −/− BMDCs (5×10 5 cells per ml) were co-cultured with 2.5×10 5 cells per ml of CD4 + CD25 high regulatory T cells in the presence of soluble anti-CD3 Ab (1 μg ml −1 ). Cells were stimulated with 10 ng ml −1 of LPS+IFNγ (10 ng ml −1 , each) for 12 h with or without anti-IL-10 neutralizing Ab (10 μg ml −1 ) in the presence or absence of membrane-permeable cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. Data are representative of two ( b – d ) to three ( a ) independent experiments. Error bars represent +s.d. Cells were stimulated in triplicate wells for each condition and s.d. was calculated from the values determined by ELISA. ** P
    Figure Legend Snippet: IFNγ antagonizes cAMP but not Treg-derived IL-10 in Socs1 −/− BMDCs. ( a ) Socs1 +/+ (5×10 5 cells per ml; black bars) and Socs1 −/− BMDCs (5×10 5 cells per ml; grey bars) were stimulated with LPS+IFNγ (10 ng ml −1 each) for 12 h in the presence or absence of graded concentrations of membrane-permeable cAMP analogue, 8-Br-cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. ( b ) Socs1 +/+ (5×10 5 cells per ml; left half of each panel) and Socs1 −/− BMDCs (5×10 5 cells per ml; right half of each panel) were stimulated with LPS (10 ng ml −1 ) for 12 h with (white bars) or without (black bars) 100 μM of 8-Br-cAMP with graded concentrations of murine recombinant IFNγ. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. IFNγ dose-dependently antagonized cAMP-mediated immunosuppression, which was enhanced by SOCS1 deficiency. ( c ) Stat1 +/+ (5×10 5 cells per ml; white bars) and Stat1 −/− (5×10 5 cells per ml; grey bars) BMDCs were stimulated with LPS (10 ng ml −1 ) for 24 h in the absence or presence of PGE2 and graded concentrations of murine recombinant IFNγ. The level of TNFα in the culture supernatant was measured by ELISA. ( d ) Socs1 +/+ and Socs1 −/− BMDCs (5×10 5 cells per ml) were co-cultured with 2.5×10 5 cells per ml of CD4 + CD25 high regulatory T cells in the presence of soluble anti-CD3 Ab (1 μg ml −1 ). Cells were stimulated with 10 ng ml −1 of LPS+IFNγ (10 ng ml −1 , each) for 12 h with or without anti-IL-10 neutralizing Ab (10 μg ml −1 ) in the presence or absence of membrane-permeable cAMP. The levels of IL-12p70 and TNFα in the culture supernatant were measured by ELISA. Data are representative of two ( b – d ) to three ( a ) independent experiments. Error bars represent +s.d. Cells were stimulated in triplicate wells for each condition and s.d. was calculated from the values determined by ELISA. ** P

    Techniques Used: Derivative Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Cell Culture

    A model for SOCS1-dependent and -independent mechanism of intestinal tolerance. Tolerance to commensal bacteria is regulated by two independent pathways: PGE2-cAMP and Treg-IL-10. Inflammatory cytokines, especially IFNγ, antagonize PGE2-cAMP-mediated suppression for Toll-like receptor signalling. SOCS1 negatively regulates IFNγ signalling, thus maintaining the PGE2-cAMP-mediated tolerance.
    Figure Legend Snippet: A model for SOCS1-dependent and -independent mechanism of intestinal tolerance. Tolerance to commensal bacteria is regulated by two independent pathways: PGE2-cAMP and Treg-IL-10. Inflammatory cytokines, especially IFNγ, antagonize PGE2-cAMP-mediated suppression for Toll-like receptor signalling. SOCS1 negatively regulates IFNγ signalling, thus maintaining the PGE2-cAMP-mediated tolerance.

    Techniques Used:

    Related Articles

    Blocking Assay:

    Article Title: Soluble CD83 Inhibits T Cell Activation by Binding to the TLR4/MD-2 Complex on CD14+ Monocytes
    Article Snippet: The following day, tubes were washed with Perm/Wash buffer (BD Biosciences). .. To set background gates, a single tube (blocked tube) containing the blocking Abs (purified anti–IL-2 [BD Biosciences], purified anti–IL-10 [eBioscience], purified anti–IFN-γ [eBioscience], and purified anti–TNF-α [BD Biosciences]) was added to the intracellular cytokine–stained blocking tube for 10 min at room temperature. .. Anti–IL-2 PE (BD Biosciences), anti–IL-10 PE-Cy7 (eBioscience), anti–IFN-γ eFluor 450 (eBioscience), and anti–TNF-α Alexa Fluor 700 (BD Biosciences) Abs were added to all tubes, and samples were incubated for 15 min at room temperature.

    Purification:

    Article Title: Soluble CD83 Inhibits T Cell Activation by Binding to the TLR4/MD-2 Complex on CD14+ Monocytes
    Article Snippet: The following day, tubes were washed with Perm/Wash buffer (BD Biosciences). .. To set background gates, a single tube (blocked tube) containing the blocking Abs (purified anti–IL-2 [BD Biosciences], purified anti–IL-10 [eBioscience], purified anti–IFN-γ [eBioscience], and purified anti–TNF-α [BD Biosciences]) was added to the intracellular cytokine–stained blocking tube for 10 min at room temperature. .. Anti–IL-2 PE (BD Biosciences), anti–IL-10 PE-Cy7 (eBioscience), anti–IFN-γ eFluor 450 (eBioscience), and anti–TNF-α Alexa Fluor 700 (BD Biosciences) Abs were added to all tubes, and samples were incubated for 15 min at room temperature.

    Staining:

    Article Title: Restricted Autoantigen Recognition Associated With Deletional and Adaptive Regulatory Mechanisms 1
    Article Snippet: Intracellular staining of cells for FoxP3 was performed using eBioscience kit (FJK.16a Ab, San Diego, CA) according to manufacturer’s instructions. .. Intracellular staining for active Caspase 3, mouse anti-IFN-γ (XMG1.2, eBioscience), and anti-IL-10 (clone JES5-16E3, eBioscience) was performed using eBioscience intracellular staining kit (cat 88-8823-88, eBioscience San Diego, CA). .. In lymph node or purified CD4+ T cell proliferation assays 1×105 lymph node cells were cultured with 2×105 3000 Rads Cs-γ irradiated splenocytes (final volume 150 µl).

    Cell Culture:

    Article Title: The Toll-like receptor 1/2 agonists Pam3CSK4 and human ?-defensin-3 differentially induce interleukin-10 and nuclear factor-?B signalling patterns in human monocytes
    Article Snippet: Three million PBMCs or purified monocytes were cultured in 1 ml medium with shBD-3 (20 μg/ml), Pam3 CSK4 (50 ng/ml), or in medium alone for 18 hr. .. In some assays PBMCs were cultured with anti-IL-10 antibody (5 μg/ml; eBioscience, San Diego, CA), anti-TLR1/2 antibodies (GD2.F4 and TLR-2.5, 10 μg/ml), or mouse IgG1 (5 or 20 μg/ml, all from Invivogen, San Diego, CA) for 1 hr before addition of TLR agonists. .. The p38 inhibitor SB-203580, CCR2 chemokine receptor antagonist RS102895 hydrochloride, were purchased from Sigma Aldrich (St Louis, MO) and used at a concentration of 10 and 100 μ m , respectively.

    Mouse Assay:

    Article Title: Positive feedback regulation between IL10 and EGFR promotes lung cancer formation
    Article Snippet: .. For inhibiting EGFR, mice, which orally administrated with Dox for 2 weeks, were intraperitoneally injected with gefitinib (20 mg/kg, twice/week; Sigma-Aldrich) for 4 weeks; mice were injected intraperitoneally with the anti-IL10 antibody (100 μg/mouse, once/week; eBioscience, Inc., San Diego, CA, USA) for 4 weeks to neutralize IL10. .. Histological staining Excised lungs were fixed with 4% formaldehyde for 48h, and embedded in paraffin for slide preparation.

    Article Title: IL-10–producing Tfh cells accumulate with age and link inflammation with age-related immune suppression
    Article Snippet: IVCCA and ELISAs IL-6 and IL-10 IVCCA was performed as previously described ( ) using biotinylated capture antibodies (Invitrogen). .. Briefly, young and aged C57BL/6 mice were intravenously injected with 10 μg of biotinylated anti–IL-6 (MP5-32C11, Invitrogen) and anti–IL-10 (JES5-16E3, Invitrogen) capture antibodies; mice were bled within 24 hours, and serum was collected. .. A luminescent ELISA was performed using anti–IL-6 (MP5-20F3, Invitrogen) or anti–IL-10 (JES5-2A5, BD Biosciences) as the coating antibody.

    Injection:

    Article Title: Positive feedback regulation between IL10 and EGFR promotes lung cancer formation
    Article Snippet: .. For inhibiting EGFR, mice, which orally administrated with Dox for 2 weeks, were intraperitoneally injected with gefitinib (20 mg/kg, twice/week; Sigma-Aldrich) for 4 weeks; mice were injected intraperitoneally with the anti-IL10 antibody (100 μg/mouse, once/week; eBioscience, Inc., San Diego, CA, USA) for 4 weeks to neutralize IL10. .. Histological staining Excised lungs were fixed with 4% formaldehyde for 48h, and embedded in paraffin for slide preparation.

    Article Title: IL-10–producing Tfh cells accumulate with age and link inflammation with age-related immune suppression
    Article Snippet: IVCCA and ELISAs IL-6 and IL-10 IVCCA was performed as previously described ( ) using biotinylated capture antibodies (Invitrogen). .. Briefly, young and aged C57BL/6 mice were intravenously injected with 10 μg of biotinylated anti–IL-6 (MP5-32C11, Invitrogen) and anti–IL-10 (JES5-16E3, Invitrogen) capture antibodies; mice were bled within 24 hours, and serum was collected. .. A luminescent ELISA was performed using anti–IL-6 (MP5-20F3, Invitrogen) or anti–IL-10 (JES5-2A5, BD Biosciences) as the coating antibody.

    Neutralization:

    Article Title: Interferons Modulate Fc?RI-dependent Production of Autoregulatory IL-10 by Circulating Human Monocytoid Dendritic Cells
    Article Snippet: Following 16 h pretreatment, IgE receptor cross-linking was then initiated by adding 50 µL of FcεRIα Ab (at 5x final concentration) to each designated well for up to 30 h incubation. .. FcεRI-mediated IL-10 neutralization experiments were performed by adding anti-IL-10 Ab (500 ng/mL, clone JESS-9D7, eBioscience) simultaneously with the AER-37 anti-FcεRIα Ab from eBioscience (1:167 of the stock or ~75 ng/mL) for 16 h. .. ELISA protocols for the detection of TNF-α, IL-10 (eBioscience), and IFN-α (in-house) have all been described previously.

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  • 94
    Thermo Fisher anti il 4
    Effects of Nup98 on Th17 cell differentiation in AVMC . CVB3 infected CD4 + T cells from peripheral blood in AVMC patients were transfected with Nup98 siRNA or Nup98 cDNA by the Human T Cell Nucleofector Kit and cultured with anti-CD3, anti-CD28, <t>anti-IL-4</t> and anti-IFN-γ for 72 h. (A) Representative pictures of Th17 cell frequencies of CD4 + T cell in pcDNA3.1, NC, pcDNA3.1-Nup98, and siRNA-nup98 groups were listed. (B) The statistical analysis for Th17 frequencies was shown in histogram. (C) The GFP-transfection efficiency was detected by flow cytometry and a typical FACS picture was shown. (D) The mRNA levels of Nup98, RORγT and IL-17 in CD4 + T cells of four groups. (E) The cultural supernatant IL-17 concentration was measured by ELISA. Data are shown as mean ± SEM. * P
    Anti Il 4, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher anti il 10 ab
    <t>IL-10</t> inhibits FcεRI-dependent TNF-α expression by blood mDCs
    Anti Il 10 Ab, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti il 10 ab/product/Thermo Fisher
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    93
    Thermo Fisher anti human il 4
    Effects of TAC in Th1-polarizing conditions Naïve CD4 + T cells were cultured under Th1-polarizing conditions (50ng ml −1 recombinant mouse IL-2, 50ng ml −1 recombinant mouse IL-12 and 10 μg ml −1 anti-mouse <t>Il-4;</t> eBioscience, CA, USA). By 96 hrs, (A) IFN-γ and ( B) IL-10 production were measured by flow cytometry and ELISA. Statistics: n ≥ 5; mean±SD; *p
    Anti Human Il 4, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti human il 4/product/Thermo Fisher
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    99
    Thermo Fisher anti il 4 neutralizing antibody
    MCs degranulation and cytokine release in response to C. albicans . (A) BMMCs degranulation in response to C. albicans was evaluated by the release of β-hexosaminidase and the synthesis of leukotrienes. No degranulation was observed after 30 min, 1 or 2 h. Data were analyzed with paired 2-way ANOVA. NS, statistically non-significant. (B) Stimulation of BMMCs with C. albicans induced a quick release of TNF-α, IL6, and IL13 (already detectable after 3 h) and of <t>IL-4.</t> C. albicans yeasts induced a more prominent cytokine release compared to hyphae. Data were analyzed with paired 2-way ANOVA. (C) Expression levels of tnf α , il6, il13, il4 were confirmed by qPCR after 3 h of co-culture. Data were analyzed by Kruskal Wallis test. Nd, non-detected. (D) Challenge of dectin-1 −/− MCs with C. albicans resulted in an impaired release of TNF-α, IL-6, and IL-13 compared to WT controls. Data were analyzed with paired 2-way ANOVA and Tukey's multiple comparison test. * p
    Anti Il 4 Neutralizing Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of Nup98 on Th17 cell differentiation in AVMC . CVB3 infected CD4 + T cells from peripheral blood in AVMC patients were transfected with Nup98 siRNA or Nup98 cDNA by the Human T Cell Nucleofector Kit and cultured with anti-CD3, anti-CD28, anti-IL-4 and anti-IFN-γ for 72 h. (A) Representative pictures of Th17 cell frequencies of CD4 + T cell in pcDNA3.1, NC, pcDNA3.1-Nup98, and siRNA-nup98 groups were listed. (B) The statistical analysis for Th17 frequencies was shown in histogram. (C) The GFP-transfection efficiency was detected by flow cytometry and a typical FACS picture was shown. (D) The mRNA levels of Nup98, RORγT and IL-17 in CD4 + T cells of four groups. (E) The cultural supernatant IL-17 concentration was measured by ELISA. Data are shown as mean ± SEM. * P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Coxsackievirus B3 Directly Induced Th17 Cell Differentiation by Inhibiting Nup98 Expression in Patients with Acute Viral Myocarditis

    doi: 10.3389/fcimb.2016.00171

    Figure Lengend Snippet: Effects of Nup98 on Th17 cell differentiation in AVMC . CVB3 infected CD4 + T cells from peripheral blood in AVMC patients were transfected with Nup98 siRNA or Nup98 cDNA by the Human T Cell Nucleofector Kit and cultured with anti-CD3, anti-CD28, anti-IL-4 and anti-IFN-γ for 72 h. (A) Representative pictures of Th17 cell frequencies of CD4 + T cell in pcDNA3.1, NC, pcDNA3.1-Nup98, and siRNA-nup98 groups were listed. (B) The statistical analysis for Th17 frequencies was shown in histogram. (C) The GFP-transfection efficiency was detected by flow cytometry and a typical FACS picture was shown. (D) The mRNA levels of Nup98, RORγT and IL-17 in CD4 + T cells of four groups. (E) The cultural supernatant IL-17 concentration was measured by ELISA. Data are shown as mean ± SEM. * P

    Article Snippet: After washing, cells were cultured with 1640 medium containing 5% FBS, 5 μg/mL of anti-CD3 (ebioscience), 2 μg/mL soluble anti-CD28 (eBioscience), 10 μg/mL anti-IL-4 (ebioscience), and 10 μg/mL anti-IFN-γ (ebioscience) for 5 days at room temperature.

    Techniques: Cell Differentiation, Infection, Transfection, Cell Culture, Flow Cytometry, Cytometry, FACS, Concentration Assay, Enzyme-linked Immunosorbent Assay

    IL-10 inhibits FcεRI-dependent TNF-α expression by blood mDCs

    Journal:

    Article Title: Interferons Modulate Fc?RI-dependent Production of Autoregulatory IL-10 by Circulating Human Monocytoid Dendritic Cells

    doi: 10.1016/j.jaci.2008.09.013

    Figure Lengend Snippet: IL-10 inhibits FcεRI-dependent TNF-α expression by blood mDCs

    Article Snippet: FcεRI-mediated IL-10 neutralization experiments were performed by adding anti-IL-10 Ab (500 ng/mL, clone JESS-9D7, eBioscience) simultaneously with the AER-37 anti-FcεRIα Ab from eBioscience (1:167 of the stock or ~75 ng/mL) for 16 h.

    Techniques: Expressing

    TLR9-induced IFN-α from pDCs enhances FcεRI-dependent IL-10 secretion by mDCs

    Journal:

    Article Title: Interferons Modulate Fc?RI-dependent Production of Autoregulatory IL-10 by Circulating Human Monocytoid Dendritic Cells

    doi: 10.1016/j.jaci.2008.09.013

    Figure Lengend Snippet: TLR9-induced IFN-α from pDCs enhances FcεRI-dependent IL-10 secretion by mDCs

    Article Snippet: FcεRI-mediated IL-10 neutralization experiments were performed by adding anti-IL-10 Ab (500 ng/mL, clone JESS-9D7, eBioscience) simultaneously with the AER-37 anti-FcεRIα Ab from eBioscience (1:167 of the stock or ~75 ng/mL) for 16 h.

    Techniques:

    Kinetics for FcεRI-dependent TNF-α and IL-10 secretion by blood mDCs

    Journal:

    Article Title: Interferons Modulate Fc?RI-dependent Production of Autoregulatory IL-10 by Circulating Human Monocytoid Dendritic Cells

    doi: 10.1016/j.jaci.2008.09.013

    Figure Lengend Snippet: Kinetics for FcεRI-dependent TNF-α and IL-10 secretion by blood mDCs

    Article Snippet: FcεRI-mediated IL-10 neutralization experiments were performed by adding anti-IL-10 Ab (500 ng/mL, clone JESS-9D7, eBioscience) simultaneously with the AER-37 anti-FcεRIα Ab from eBioscience (1:167 of the stock or ~75 ng/mL) for 16 h.

    Techniques:

    Inhibitory effects of IL-10 on FcεRI-dependent responses in mDCs are not from a down-regulation in FcεRIα and co-stimulatory molecule expression

    Journal:

    Article Title: Interferons Modulate Fc?RI-dependent Production of Autoregulatory IL-10 by Circulating Human Monocytoid Dendritic Cells

    doi: 10.1016/j.jaci.2008.09.013

    Figure Lengend Snippet: Inhibitory effects of IL-10 on FcεRI-dependent responses in mDCs are not from a down-regulation in FcεRIα and co-stimulatory molecule expression

    Article Snippet: FcεRI-mediated IL-10 neutralization experiments were performed by adding anti-IL-10 Ab (500 ng/mL, clone JESS-9D7, eBioscience) simultaneously with the AER-37 anti-FcεRIα Ab from eBioscience (1:167 of the stock or ~75 ng/mL) for 16 h.

    Techniques: Expressing

    Interferons modulate FcεRI-dependent IL-10 secretion by mDCs

    Journal:

    Article Title: Interferons Modulate Fc?RI-dependent Production of Autoregulatory IL-10 by Circulating Human Monocytoid Dendritic Cells

    doi: 10.1016/j.jaci.2008.09.013

    Figure Lengend Snippet: Interferons modulate FcεRI-dependent IL-10 secretion by mDCs

    Article Snippet: FcεRI-mediated IL-10 neutralization experiments were performed by adding anti-IL-10 Ab (500 ng/mL, clone JESS-9D7, eBioscience) simultaneously with the AER-37 anti-FcεRIα Ab from eBioscience (1:167 of the stock or ~75 ng/mL) for 16 h.

    Techniques:

    Effects of TAC in Th1-polarizing conditions Naïve CD4 + T cells were cultured under Th1-polarizing conditions (50ng ml −1 recombinant mouse IL-2, 50ng ml −1 recombinant mouse IL-12 and 10 μg ml −1 anti-mouse Il-4; eBioscience, CA, USA). By 96 hrs, (A) IFN-γ and ( B) IL-10 production were measured by flow cytometry and ELISA. Statistics: n ≥ 5; mean±SD; *p

    Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

    Article Title: Age-dependent metabolic and immunosuppressive effects of Tacrolimus

    doi: 10.1111/ajt.14087

    Figure Lengend Snippet: Effects of TAC in Th1-polarizing conditions Naïve CD4 + T cells were cultured under Th1-polarizing conditions (50ng ml −1 recombinant mouse IL-2, 50ng ml −1 recombinant mouse IL-12 and 10 μg ml −1 anti-mouse Il-4; eBioscience, CA, USA). By 96 hrs, (A) IFN-γ and ( B) IL-10 production were measured by flow cytometry and ELISA. Statistics: n ≥ 5; mean±SD; *p

    Article Snippet: The following antibodies were used for murine cells: 50ng ml−1 recombinant mouse IL-2, 50ng ml−1 recombinant mouse IL-12 and 10 μg ml−1 anti-mouse IL-4 (all eBioscience, CA, USA); for human cells we used: 50ng ml−1 recombinant human IL-2, 50ng ml−1 recombinant human IL-12 and 10 μg ml−1 anti-human IL-4 (all eBioscience, CA, USA).

    Techniques: Cell Culture, Recombinant, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    MCs degranulation and cytokine release in response to C. albicans . (A) BMMCs degranulation in response to C. albicans was evaluated by the release of β-hexosaminidase and the synthesis of leukotrienes. No degranulation was observed after 30 min, 1 or 2 h. Data were analyzed with paired 2-way ANOVA. NS, statistically non-significant. (B) Stimulation of BMMCs with C. albicans induced a quick release of TNF-α, IL6, and IL13 (already detectable after 3 h) and of IL-4. C. albicans yeasts induced a more prominent cytokine release compared to hyphae. Data were analyzed with paired 2-way ANOVA. (C) Expression levels of tnf α , il6, il13, il4 were confirmed by qPCR after 3 h of co-culture. Data were analyzed by Kruskal Wallis test. Nd, non-detected. (D) Challenge of dectin-1 −/− MCs with C. albicans resulted in an impaired release of TNF-α, IL-6, and IL-13 compared to WT controls. Data were analyzed with paired 2-way ANOVA and Tukey's multiple comparison test. * p

    Journal: Frontiers in Immunology

    Article Title: Mast Cells Respond to Candida albicans Infections and Modulate Macrophages Phagocytosis of the Fungus

    doi: 10.3389/fimmu.2018.02829

    Figure Lengend Snippet: MCs degranulation and cytokine release in response to C. albicans . (A) BMMCs degranulation in response to C. albicans was evaluated by the release of β-hexosaminidase and the synthesis of leukotrienes. No degranulation was observed after 30 min, 1 or 2 h. Data were analyzed with paired 2-way ANOVA. NS, statistically non-significant. (B) Stimulation of BMMCs with C. albicans induced a quick release of TNF-α, IL6, and IL13 (already detectable after 3 h) and of IL-4. C. albicans yeasts induced a more prominent cytokine release compared to hyphae. Data were analyzed with paired 2-way ANOVA. (C) Expression levels of tnf α , il6, il13, il4 were confirmed by qPCR after 3 h of co-culture. Data were analyzed by Kruskal Wallis test. Nd, non-detected. (D) Challenge of dectin-1 −/− MCs with C. albicans resulted in an impaired release of TNF-α, IL-6, and IL-13 compared to WT controls. Data were analyzed with paired 2-way ANOVA and Tukey's multiple comparison test. * p

    Article Snippet: In some experiment, 50 pg·ml−1 recombinant IL-4 (Peprotech), 100 pg·ml−1 recombinant TNF-α (Immunotools), 10 μg·ml−1 anti-IL-4 neutralizing antibody (11B11, eBioscience), 10 μg·ml−1 anti-TNF-α neutralizing antibody (MP6-XT22, Miltenyi Biotec), or conditioned media were used to stimulate peritoneal macrophages together with BMMCs and C. albicans .

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Co-Culture Assay

    MCs activation influence fungal clearance by peritoneal macrophages. (A) Gating strategy used for the evaluation of macrophages phagocytosis. (B) C. albicans phagocytosis by macrophages is impaired by the presence of naïve (resting) MCs. The addition of exogenous IL-4 nor its neutralization with monoclonal antibodies (C) as well as the addition of TNF-α nor its neutralization (D) restored or inhibited macrophages phagocytosis. Phagocytosis was assessed by flow cytometry after 1 h of co-culture and the phagocytosis index expressed as the fold-change over the phagocytosis percentage of macrophages stimulated with C. albicans alone. (E) Similarly, the presence of conditioned media (C.M.) to resting MCs did not affect macrophages phagocytosis ability. Data were analyzed with Kruscal-Wallis and Dunn's multiple comparison tests. (F) Immunofluorescence analyses of macrophages-BMMCs- C. albicans co-cultures indicate that MCs and macrophages interact during the process of phagocytosis. Scale bar: 10 μm. * p

    Journal: Frontiers in Immunology

    Article Title: Mast Cells Respond to Candida albicans Infections and Modulate Macrophages Phagocytosis of the Fungus

    doi: 10.3389/fimmu.2018.02829

    Figure Lengend Snippet: MCs activation influence fungal clearance by peritoneal macrophages. (A) Gating strategy used for the evaluation of macrophages phagocytosis. (B) C. albicans phagocytosis by macrophages is impaired by the presence of naïve (resting) MCs. The addition of exogenous IL-4 nor its neutralization with monoclonal antibodies (C) as well as the addition of TNF-α nor its neutralization (D) restored or inhibited macrophages phagocytosis. Phagocytosis was assessed by flow cytometry after 1 h of co-culture and the phagocytosis index expressed as the fold-change over the phagocytosis percentage of macrophages stimulated with C. albicans alone. (E) Similarly, the presence of conditioned media (C.M.) to resting MCs did not affect macrophages phagocytosis ability. Data were analyzed with Kruscal-Wallis and Dunn's multiple comparison tests. (F) Immunofluorescence analyses of macrophages-BMMCs- C. albicans co-cultures indicate that MCs and macrophages interact during the process of phagocytosis. Scale bar: 10 μm. * p

    Article Snippet: In some experiment, 50 pg·ml−1 recombinant IL-4 (Peprotech), 100 pg·ml−1 recombinant TNF-α (Immunotools), 10 μg·ml−1 anti-IL-4 neutralizing antibody (11B11, eBioscience), 10 μg·ml−1 anti-TNF-α neutralizing antibody (MP6-XT22, Miltenyi Biotec), or conditioned media were used to stimulate peritoneal macrophages together with BMMCs and C. albicans .

    Techniques: Activation Assay, Neutralization, Flow Cytometry, Cytometry, Co-Culture Assay, Immunofluorescence