goat anti mouse il 17a  (R&D Systems)

 
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    Name:
    Mouse IL 17 IL 17A Antibody
    Description:
    The Mouse IL 17 IL 17A Antibody from R D Systems is a goat polyclonal antibody to IL 17 IL 17A This antibody reacts with mouse The Mouse IL 17 IL 17A Antibody has been validated for the following applications Western Blot Neutralization Intracellular Staining by Flow Cytometry CyTOF ready
    Catalog Number:
    AF-421-NA
    Price:
    375
    Category:
    Primary Antibodies
    Applications:
    Western Blot, Neutralization, Intracellular Staining by Flow Cytometry, CyTOF-ready
    Purity:
    Immunogen affinity purified
    Conjugate:
    Unconjugated
    Immunogen:
    E. coli-derived recombinant mouse IL-17, Thr22-Ala158, Accession # Q62386
    Size:
    100 ug
    Host:
    Goat
    Isotype:
    IgG
    Buy from Supplier


    Structured Review

    R&D Systems goat anti mouse il 17a
    Mouse IL 17 IL 17A Antibody
    The Mouse IL 17 IL 17A Antibody from R D Systems is a goat polyclonal antibody to IL 17 IL 17A This antibody reacts with mouse The Mouse IL 17 IL 17A Antibody has been validated for the following applications Western Blot Neutralization Intracellular Staining by Flow Cytometry CyTOF ready
    https://www.bioz.com/result/goat anti mouse il 17a/product/R&D Systems
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse il 17a - by Bioz Stars, 2021-09
    93/100 stars

    Images

    1) Product Images from "Evidence for anti-inflammatory effects of C5a on the innate IL-17A/IL-23 axis"

    Article Title: Evidence for anti-inflammatory effects of C5a on the innate IL-17A/IL-23 axis

    Journal: The FASEB Journal

    doi: 10.1096/fj.11-199216

    Neutralizing of IL-17A or IL-23 (p19) improves survival after endotoxemia. A ) Time course of IL-23 (p19) in plasma after LPS challenge of C57BL/6J mice in vivo ( n =5/group), ELISA. B ) Time course of IL-17A in plasma after LPS challenge in vivo ( n =5/group).
    Figure Legend Snippet: Neutralizing of IL-17A or IL-23 (p19) improves survival after endotoxemia. A ) Time course of IL-23 (p19) in plasma after LPS challenge of C57BL/6J mice in vivo ( n =5/group), ELISA. B ) Time course of IL-17A in plasma after LPS challenge in vivo ( n =5/group).

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

    Ability of C5a to suppress IL-17A production in macrophages. A ) PEMs (Wt) were incubated with LPS (1 μg/ml) in the copresence of the indicated concentrations of recombinant mouse C5a. Concentrations of IL-17A (ELISA) were detected in cell culture
    Figure Legend Snippet: Ability of C5a to suppress IL-17A production in macrophages. A ) PEMs (Wt) were incubated with LPS (1 μg/ml) in the copresence of the indicated concentrations of recombinant mouse C5a. Concentrations of IL-17A (ELISA) were detected in cell culture

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

    Characterization of IL-17A release by macrophages. A ) Flow cytometry analysis for intracellular cytokine staining for IL-17A as a function of the macrophage surface markers, CD11b and F4/80. B ) IL-17A release from LPS-activated PEMs from C57BL/6J mice
    Figure Legend Snippet: Characterization of IL-17A release by macrophages. A ) Flow cytometry analysis for intracellular cytokine staining for IL-17A as a function of the macrophage surface markers, CD11b and F4/80. B ) IL-17A release from LPS-activated PEMs from C57BL/6J mice

    Techniques Used: Flow Cytometry, Cytometry, Staining, Mouse Assay

    IL-17A release by macrophages from different origins. A ) PEMs from C57BL/6J (Wt) mice were isolated from the peritoneum after elicitation with thioglycollate and stained after Wright-Giemsa. B ) Production of IL-17A by PEMs incubated for 10 h at 37°C
    Figure Legend Snippet: IL-17A release by macrophages from different origins. A ) PEMs from C57BL/6J (Wt) mice were isolated from the peritoneum after elicitation with thioglycollate and stained after Wright-Giemsa. B ) Production of IL-17A by PEMs incubated for 10 h at 37°C

    Techniques Used: Mouse Assay, Isolation, Staining, Incubation

    C5a suppresses the IL-17A/IL-23 axis via enhancement of IL-10. A ) ELISA of IL-10 release after incubation with LPS (1 μg/ml, 10 h) in PEMs from C57BL/6J mice in the absence or copresence of recombinant C5a or C5a alone. B ) Time course of IL-10
    Figure Legend Snippet: C5a suppresses the IL-17A/IL-23 axis via enhancement of IL-10. A ) ELISA of IL-10 release after incubation with LPS (1 μg/ml, 10 h) in PEMs from C57BL/6J mice in the absence or copresence of recombinant C5a or C5a alone. B ) Time course of IL-10

    Techniques Used: Enzyme-linked Immunosorbent Assay, Incubation, Mouse Assay, Recombinant

    Related Articles

    Incubation:

    Article Title: Development of an IL-17A DNA Vaccine to Treat Systemic Lupus Erythematosus in Mice
    Article Snippet: .. The blots were incubated with sera from mice immunized with the IL-17A1 or IL-17A2 vaccine, an anti-BSA antibody, or a commercially available anti-IL-17A antibody (AF-421-NA, R & D Systems). ..

    Mouse Assay:

    Article Title: Development of an IL-17A DNA Vaccine to Treat Systemic Lupus Erythematosus in Mice
    Article Snippet: .. The blots were incubated with sera from mice immunized with the IL-17A1 or IL-17A2 vaccine, an anti-BSA antibody, or a commercially available anti-IL-17A antibody (AF-421-NA, R & D Systems). ..

    Article Title: IL-33 signaling regulates innate IL-17A and IL-22 production via suppression of PGE2 during lung fungal infection
    Article Snippet: .. For in vivo IL-17A and IL-22 neutralization, Ilrl1 −/− mice were challenged intratracheally with 7 × 107 A. fumigatus conidia in 50 µl and 24 h thereafter, mice were administered 50 µg of rat anti-mouse IL-17A (Cat. #MAB421) or IL-22 (Cat. #MAB5821) or rat IgG isotype control antibody or polyclonal goat anti-mouse IL-17A (Cat. #AF-421-NA) or IL-22 (Cat. #AF582) or goat IgG isotype control antibody (Cat. #AB-108-C) (all antibodies from R & D Systems) intratracheally. ..

    In Vivo:

    Article Title: IL-33 signaling regulates innate IL-17A and IL-22 production via suppression of PGE2 during lung fungal infection
    Article Snippet: .. For in vivo IL-17A and IL-22 neutralization, Ilrl1 −/− mice were challenged intratracheally with 7 × 107 A. fumigatus conidia in 50 µl and 24 h thereafter, mice were administered 50 µg of rat anti-mouse IL-17A (Cat. #MAB421) or IL-22 (Cat. #MAB5821) or rat IgG isotype control antibody or polyclonal goat anti-mouse IL-17A (Cat. #AF-421-NA) or IL-22 (Cat. #AF582) or goat IgG isotype control antibody (Cat. #AB-108-C) (all antibodies from R & D Systems) intratracheally. ..

    Article Title: Enhanced IL-17 signalling following myocardial ischaemia/reperfusion injury
    Article Snippet: .. 2.2 In vivo neutralization of IL-17A In a different set of experiments rats were treated with 200 μg of anti-IL-17A polyclonal antibody intraperitoneally (R & D Systems, AF-421-NA) or PBS 2 h before undergoing in vivo ischemia/reperfusion injury. ..

    Neutralization:

    Article Title: IL-33 signaling regulates innate IL-17A and IL-22 production via suppression of PGE2 during lung fungal infection
    Article Snippet: .. For in vivo IL-17A and IL-22 neutralization, Ilrl1 −/− mice were challenged intratracheally with 7 × 107 A. fumigatus conidia in 50 µl and 24 h thereafter, mice were administered 50 µg of rat anti-mouse IL-17A (Cat. #MAB421) or IL-22 (Cat. #MAB5821) or rat IgG isotype control antibody or polyclonal goat anti-mouse IL-17A (Cat. #AF-421-NA) or IL-22 (Cat. #AF582) or goat IgG isotype control antibody (Cat. #AB-108-C) (all antibodies from R & D Systems) intratracheally. ..

    Article Title: Enhanced IL-17 signalling following myocardial ischaemia/reperfusion injury
    Article Snippet: .. 2.2 In vivo neutralization of IL-17A In a different set of experiments rats were treated with 200 μg of anti-IL-17A polyclonal antibody intraperitoneally (R & D Systems, AF-421-NA) or PBS 2 h before undergoing in vivo ischemia/reperfusion injury. ..

    Flow Cytometry:

    Article Title: MHC class II alleles associated with Th1 rather than Th17 type immunity drive the onset of early arthritis in a rat model of rheumatoid arthritis
    Article Snippet: .. Antibodies for flow cytometry The following FITC, Alexa Fluor 488, PE, Pe‐Cy5, PerCP‐Cy5.5, APC, Alexa Fluor 648, APC‐Cy7, Qdot‐655 and biotin conjugated antibodies were used for flow cytometry: CD4 (OX‐35), CD8a (OX‐8), CD25 (OX‐39), CD45RC (OX‐22), and CD3 (1F4) were purchased from BD Pharmingen (San Diego, CA); IFN‐γ (DB‐1), CD45 (OX‐1), CD90/Thy1 (OX7), TCR Vβ8.2/8.4 (R78) and αβTCR (R73) were purchased from BioLegend (San Diego, CA); TCR Vβ16.1 (His42) and TCR Vβ10 (G101) were purchased unlabeled from LSBio (Seattle, WA, USA) and used with a secondary polyclonal APC‐Cy7‐labelled goat anti‐mouse antibody (BioLegend); Foxp3 (FJK‐16s), Ki67 (SolA15) and CD11b/c (OX42) were purchased from eBioscience (San Diego, CA); Polyclonal goat‐anti mouse IL‐17 was purchased from R & D Systems (IC421P). ..

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  • 93
    R&D Systems goat anti mouse il 17
    Tbet deficiency prevents induction of transplantation tolerance by combined co-stimulation blockade with persistent T17/Th2 skewing of alloantigen specific cytokine profile, and PMN, CD4, and <t>IL-17-producing</t> CD8 T cell infiltration. ( A ) Kaplan-Meier survival
    Goat Anti Mouse Il 17, supplied by R&D Systems, 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/goat anti mouse il 17/product/R&D Systems
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse il 17 - by Bioz Stars, 2021-09
    93/100 stars
      Buy from Supplier

    92
    R&D Systems anti il 17
    5-FU inhibition is rescued by thymidine A. Naïve CD4 + T cells from C57BL/6 mice were differentiated under T H 17 polarizing conditions in the presence of 5-FU (1.0 μM) and thymidine (1.0 μM) for 3 days and then were re-stimulated with PMA/ionomycin for 5 hours, stained for intracellular <t>IL-17,</t> and analyzed by flow cytometry as shown. Supernatants from the cells were saved and tested via ELISA. The cells cultured for 48 hours and mRNA expression of the indicated genes was determined by qPCR. B. Repetition of (A) using T H 1 polarizing conditions. C. Repetition of (A) and (B), except using T H 2 polarizing conditions. D. Repetition of (A), (B), and (C) with Treg polarizing conditions instead. ** p
    Anti Il 17, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti il 17/product/R&D Systems
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti il 17 - by Bioz Stars, 2021-09
    92/100 stars
      Buy from Supplier

    97
    R&D Systems goat anti mouse il 17 neutralizing antibody
    IL-9 strengthens the survival and protumor effect of mast cells in tumor microenvironment. (A and B) Mast cells upregulated the expressions of IL-9 by Treg cells. BMMCs were injected into tumor-bearing mice with <t>IL-17</t> antibody or control antibody. Seven days later, the tumor-infiltrating Treg cells were isolated for the analysis of IL-9 by real time RT-PCR (A). Or the isolated Treg cells were cultured for 48 hours. The supernatant was used for IL-9 ELISA assay (B). (C and D) The interference of IL-9 signaling affected mast cell-mediated MDSC infiltration and mast cell-promoted tumor growth. 5×10 6 BMMCs were injected into tumor-bearing mice (n = 6) by i.v. injection. IL-9 neutralizing antibody or CD25 depleting antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 12, the tumor-infiltrating lymphocytes were used to analyze Gr-1 + CD11b + MDSCs by flow cytometry (C), and the tumor growth was monitored by measuring the length (L) and width (W) of tumors. The volume (V) of the tumor was calculated by the formula V = (L×W2)/2 (D). (E) IL-9 affected the survival of mast cells in tumor microenvironment. 1×10 6 CFSE-labeled BMMCs were directly injected into tumor tissue with multiple injection sites. IL-9 neutralizing antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. The tumor tissues were surgically excised on day 7 for fluorescent analysis of frozen sections.
    Goat Anti Mouse Il 17 Neutralizing Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse il 17 neutralizing antibody/product/R&D Systems
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse il 17 neutralizing antibody - by Bioz Stars, 2021-09
    97/100 stars
      Buy from Supplier

    Image Search Results


    Tbet deficiency prevents induction of transplantation tolerance by combined co-stimulation blockade with persistent T17/Th2 skewing of alloantigen specific cytokine profile, and PMN, CD4, and IL-17-producing CD8 T cell infiltration. ( A ) Kaplan-Meier survival

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Targeting Tim-1 to overcome resistance to transplantation tolerance mediated by CD8 T17 cells

    doi: 10.1073/pnas.0812538106

    Figure Lengend Snippet: Tbet deficiency prevents induction of transplantation tolerance by combined co-stimulation blockade with persistent T17/Th2 skewing of alloantigen specific cytokine profile, and PMN, CD4, and IL-17-producing CD8 T cell infiltration. ( A ) Kaplan-Meier survival

    Article Snippet: Frozen sections were used for immunofluorescence staining using goat anti-mouse IL-17 (R & D Systems), rat anti-Mouse CD4 and CD8 (both from BioExpress) as primary antibodies.

    Techniques: Transplantation Assay

    In vivo IL-17 neutralization inhibits rejection and facilitates allograft survival with combined co-stimulation blockade in Tbet KO recipients. ( A ) Fully mismatched cardiac allograft survival in Tbet KO recipients treated with CTLA4Ig+MR1 and anti-IL-17

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Targeting Tim-1 to overcome resistance to transplantation tolerance mediated by CD8 T17 cells

    doi: 10.1073/pnas.0812538106

    Figure Lengend Snippet: In vivo IL-17 neutralization inhibits rejection and facilitates allograft survival with combined co-stimulation blockade in Tbet KO recipients. ( A ) Fully mismatched cardiac allograft survival in Tbet KO recipients treated with CTLA4Ig+MR1 and anti-IL-17

    Article Snippet: Frozen sections were used for immunofluorescence staining using goat anti-mouse IL-17 (R & D Systems), rat anti-Mouse CD4 and CD8 (both from BioExpress) as primary antibodies.

    Techniques: In Vivo, Neutralization

    Neutralizing of IL-17A or IL-23 (p19) improves survival after endotoxemia. A ) Time course of IL-23 (p19) in plasma after LPS challenge of C57BL/6J mice in vivo ( n =5/group), ELISA. B ) Time course of IL-17A in plasma after LPS challenge in vivo ( n =5/group).

    Journal: The FASEB Journal

    Article Title: Evidence for anti-inflammatory effects of C5a on the innate IL-17A/IL-23 axis

    doi: 10.1096/fj.11-199216

    Figure Lengend Snippet: Neutralizing of IL-17A or IL-23 (p19) improves survival after endotoxemia. A ) Time course of IL-23 (p19) in plasma after LPS challenge of C57BL/6J mice in vivo ( n =5/group), ELISA. B ) Time course of IL-17A in plasma after LPS challenge in vivo ( n =5/group).

    Article Snippet: Antibodies used were goat anti-mouse IL-17A (R & D Systems), rat anti-mouse F4/80 (eBioscience), rabbit anti-goat biotinylated IgG, streptavidin AF488, and rabbit anti-rat AF594 (all from Invitrogen).

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

    Ability of C5a to suppress IL-17A production in macrophages. A ) PEMs (Wt) were incubated with LPS (1 μg/ml) in the copresence of the indicated concentrations of recombinant mouse C5a. Concentrations of IL-17A (ELISA) were detected in cell culture

    Journal: The FASEB Journal

    Article Title: Evidence for anti-inflammatory effects of C5a on the innate IL-17A/IL-23 axis

    doi: 10.1096/fj.11-199216

    Figure Lengend Snippet: Ability of C5a to suppress IL-17A production in macrophages. A ) PEMs (Wt) were incubated with LPS (1 μg/ml) in the copresence of the indicated concentrations of recombinant mouse C5a. Concentrations of IL-17A (ELISA) were detected in cell culture

    Article Snippet: Antibodies used were goat anti-mouse IL-17A (R & D Systems), rat anti-mouse F4/80 (eBioscience), rabbit anti-goat biotinylated IgG, streptavidin AF488, and rabbit anti-rat AF594 (all from Invitrogen).

    Techniques: Incubation, Recombinant, Enzyme-linked Immunosorbent Assay, Cell Culture

    Characterization of IL-17A release by macrophages. A ) Flow cytometry analysis for intracellular cytokine staining for IL-17A as a function of the macrophage surface markers, CD11b and F4/80. B ) IL-17A release from LPS-activated PEMs from C57BL/6J mice

    Journal: The FASEB Journal

    Article Title: Evidence for anti-inflammatory effects of C5a on the innate IL-17A/IL-23 axis

    doi: 10.1096/fj.11-199216

    Figure Lengend Snippet: Characterization of IL-17A release by macrophages. A ) Flow cytometry analysis for intracellular cytokine staining for IL-17A as a function of the macrophage surface markers, CD11b and F4/80. B ) IL-17A release from LPS-activated PEMs from C57BL/6J mice

    Article Snippet: Antibodies used were goat anti-mouse IL-17A (R & D Systems), rat anti-mouse F4/80 (eBioscience), rabbit anti-goat biotinylated IgG, streptavidin AF488, and rabbit anti-rat AF594 (all from Invitrogen).

    Techniques: Flow Cytometry, Cytometry, Staining, Mouse Assay

    IL-17A release by macrophages from different origins. A ) PEMs from C57BL/6J (Wt) mice were isolated from the peritoneum after elicitation with thioglycollate and stained after Wright-Giemsa. B ) Production of IL-17A by PEMs incubated for 10 h at 37°C

    Journal: The FASEB Journal

    Article Title: Evidence for anti-inflammatory effects of C5a on the innate IL-17A/IL-23 axis

    doi: 10.1096/fj.11-199216

    Figure Lengend Snippet: IL-17A release by macrophages from different origins. A ) PEMs from C57BL/6J (Wt) mice were isolated from the peritoneum after elicitation with thioglycollate and stained after Wright-Giemsa. B ) Production of IL-17A by PEMs incubated for 10 h at 37°C

    Article Snippet: Antibodies used were goat anti-mouse IL-17A (R & D Systems), rat anti-mouse F4/80 (eBioscience), rabbit anti-goat biotinylated IgG, streptavidin AF488, and rabbit anti-rat AF594 (all from Invitrogen).

    Techniques: Mouse Assay, Isolation, Staining, Incubation

    C5a suppresses the IL-17A/IL-23 axis via enhancement of IL-10. A ) ELISA of IL-10 release after incubation with LPS (1 μg/ml, 10 h) in PEMs from C57BL/6J mice in the absence or copresence of recombinant C5a or C5a alone. B ) Time course of IL-10

    Journal: The FASEB Journal

    Article Title: Evidence for anti-inflammatory effects of C5a on the innate IL-17A/IL-23 axis

    doi: 10.1096/fj.11-199216

    Figure Lengend Snippet: C5a suppresses the IL-17A/IL-23 axis via enhancement of IL-10. A ) ELISA of IL-10 release after incubation with LPS (1 μg/ml, 10 h) in PEMs from C57BL/6J mice in the absence or copresence of recombinant C5a or C5a alone. B ) Time course of IL-10

    Article Snippet: Antibodies used were goat anti-mouse IL-17A (R & D Systems), rat anti-mouse F4/80 (eBioscience), rabbit anti-goat biotinylated IgG, streptavidin AF488, and rabbit anti-rat AF594 (all from Invitrogen).

    Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Mouse Assay, Recombinant

    5-FU inhibition is rescued by thymidine A. Naïve CD4 + T cells from C57BL/6 mice were differentiated under T H 17 polarizing conditions in the presence of 5-FU (1.0 μM) and thymidine (1.0 μM) for 3 days and then were re-stimulated with PMA/ionomycin for 5 hours, stained for intracellular IL-17, and analyzed by flow cytometry as shown. Supernatants from the cells were saved and tested via ELISA. The cells cultured for 48 hours and mRNA expression of the indicated genes was determined by qPCR. B. Repetition of (A) using T H 1 polarizing conditions. C. Repetition of (A) and (B), except using T H 2 polarizing conditions. D. Repetition of (A), (B), and (C) with Treg polarizing conditions instead. ** p

    Journal: Oncotarget

    Article Title: 5-Fluorouracil targets thymidylate synthase in the selective suppression of TH17 cell differentiation

    doi: 10.18632/oncotarget.8344

    Figure Lengend Snippet: 5-FU inhibition is rescued by thymidine A. Naïve CD4 + T cells from C57BL/6 mice were differentiated under T H 17 polarizing conditions in the presence of 5-FU (1.0 μM) and thymidine (1.0 μM) for 3 days and then were re-stimulated with PMA/ionomycin for 5 hours, stained for intracellular IL-17, and analyzed by flow cytometry as shown. Supernatants from the cells were saved and tested via ELISA. The cells cultured for 48 hours and mRNA expression of the indicated genes was determined by qPCR. B. Repetition of (A) using T H 1 polarizing conditions. C. Repetition of (A) and (B), except using T H 2 polarizing conditions. D. Repetition of (A), (B), and (C) with Treg polarizing conditions instead. ** p

    Article Snippet: Cytokine ELISA Supernatants from cell cultures were collected after activation under various conditions and secreted cytokines in the supernatants were measured by ELISA kits with purified coating and biotinylated detection antibodies: anti-IL-17, anti-IFN-γ (R & D systems), anti-IL-9, anti-IL-22, anti-IL-1β (e Bioscience) and anti-IL-4 (BD Bioscience).

    Techniques: Inhibition, Mouse Assay, Staining, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    5-FU alters STAT3 DNA binding activity in T H 17 cells A. Naïve CD4 + T cells from C57BL/B6 mice cultured in T H 17 polarizing conditions in the presence of 5-FU (1.0 μM) for 3 days prior to western blotting for RORγt and other indicated proteins. B. The same cells were cultured for 60 hours, followed by ChIP assay. 3 μg of anti- RORγt antibody or isotype-matched IgG control antibody were used in the immunoprecipitation step. qPCR was used to quantify the amount of precipitated DNA with primers flanking the RORγt binding the CNS2 of the IL-17 promoter region. C. 293T cells transfected with STAT3 plasmid for 40 hours in the presence of 5-FU (1 μM). The cytosolic fraction and nuclear fraction proteins were analyzed via western blotting. Cells were cultured as in (A), with western blotting for STAT3 protein phosphorylation and STAT3 protein expression at the times indicated. Cells treated as in (A) analyzed via ChIP assay. 3 μg of anti-STAT3 antibody or isotype-matched IgG as control antibody were used in the immunoprecipitation step. qPCR was used to quantify the amount of precipitated DNA with primers flanking the STAT3 binding site of the RORγt D. and the CNS2 of the IL-17 (E) promoter region. * p

    Journal: Oncotarget

    Article Title: 5-Fluorouracil targets thymidylate synthase in the selective suppression of TH17 cell differentiation

    doi: 10.18632/oncotarget.8344

    Figure Lengend Snippet: 5-FU alters STAT3 DNA binding activity in T H 17 cells A. Naïve CD4 + T cells from C57BL/B6 mice cultured in T H 17 polarizing conditions in the presence of 5-FU (1.0 μM) for 3 days prior to western blotting for RORγt and other indicated proteins. B. The same cells were cultured for 60 hours, followed by ChIP assay. 3 μg of anti- RORγt antibody or isotype-matched IgG control antibody were used in the immunoprecipitation step. qPCR was used to quantify the amount of precipitated DNA with primers flanking the RORγt binding the CNS2 of the IL-17 promoter region. C. 293T cells transfected with STAT3 plasmid for 40 hours in the presence of 5-FU (1 μM). The cytosolic fraction and nuclear fraction proteins were analyzed via western blotting. Cells were cultured as in (A), with western blotting for STAT3 protein phosphorylation and STAT3 protein expression at the times indicated. Cells treated as in (A) analyzed via ChIP assay. 3 μg of anti-STAT3 antibody or isotype-matched IgG as control antibody were used in the immunoprecipitation step. qPCR was used to quantify the amount of precipitated DNA with primers flanking the STAT3 binding site of the RORγt D. and the CNS2 of the IL-17 (E) promoter region. * p

    Article Snippet: Cytokine ELISA Supernatants from cell cultures were collected after activation under various conditions and secreted cytokines in the supernatants were measured by ELISA kits with purified coating and biotinylated detection antibodies: anti-IL-17, anti-IFN-γ (R & D systems), anti-IL-9, anti-IL-22, anti-IL-1β (e Bioscience) and anti-IL-4 (BD Bioscience).

    Techniques: Binding Assay, Activity Assay, Mouse Assay, Cell Culture, Western Blot, Chromatin Immunoprecipitation, Immunoprecipitation, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Expressing

    Knockdown of the TS suppresses T H 17 and T H 1 cell differentiation A. Naïve CD4 + T cells from C57BL/6 mice were transfected with TS siRNA or control siRNA and differentiated under T H 17 polarizing conditions for 72 hours. Cell lysates were analyzed by western blotting. The cells were cultured for 72 hours and then were re-stimulated with PMA/ionomycin for 5 hours, stained for intracellular IL-17, and analyzed by flow cytometry, and the supernatants were analyzed for IL-17 by ELISA. *** p

    Journal: Oncotarget

    Article Title: 5-Fluorouracil targets thymidylate synthase in the selective suppression of TH17 cell differentiation

    doi: 10.18632/oncotarget.8344

    Figure Lengend Snippet: Knockdown of the TS suppresses T H 17 and T H 1 cell differentiation A. Naïve CD4 + T cells from C57BL/6 mice were transfected with TS siRNA or control siRNA and differentiated under T H 17 polarizing conditions for 72 hours. Cell lysates were analyzed by western blotting. The cells were cultured for 72 hours and then were re-stimulated with PMA/ionomycin for 5 hours, stained for intracellular IL-17, and analyzed by flow cytometry, and the supernatants were analyzed for IL-17 by ELISA. *** p

    Article Snippet: Cytokine ELISA Supernatants from cell cultures were collected after activation under various conditions and secreted cytokines in the supernatants were measured by ELISA kits with purified coating and biotinylated detection antibodies: anti-IL-17, anti-IFN-γ (R & D systems), anti-IL-9, anti-IL-22, anti-IL-1β (e Bioscience) and anti-IL-4 (BD Bioscience).

    Techniques: Cell Differentiation, Mouse Assay, Transfection, Western Blot, Cell Culture, Staining, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    IL-9 strengthens the survival and protumor effect of mast cells in tumor microenvironment. (A and B) Mast cells upregulated the expressions of IL-9 by Treg cells. BMMCs were injected into tumor-bearing mice with IL-17 antibody or control antibody. Seven days later, the tumor-infiltrating Treg cells were isolated for the analysis of IL-9 by real time RT-PCR (A). Or the isolated Treg cells were cultured for 48 hours. The supernatant was used for IL-9 ELISA assay (B). (C and D) The interference of IL-9 signaling affected mast cell-mediated MDSC infiltration and mast cell-promoted tumor growth. 5×10 6 BMMCs were injected into tumor-bearing mice (n = 6) by i.v. injection. IL-9 neutralizing antibody or CD25 depleting antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 12, the tumor-infiltrating lymphocytes were used to analyze Gr-1 + CD11b + MDSCs by flow cytometry (C), and the tumor growth was monitored by measuring the length (L) and width (W) of tumors. The volume (V) of the tumor was calculated by the formula V = (L×W2)/2 (D). (E) IL-9 affected the survival of mast cells in tumor microenvironment. 1×10 6 CFSE-labeled BMMCs were directly injected into tumor tissue with multiple injection sites. IL-9 neutralizing antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. The tumor tissues were surgically excised on day 7 for fluorescent analysis of frozen sections.

    Journal: PLoS ONE

    Article Title: Mast Cells Mobilize Myeloid-Derived Suppressor Cells and Treg Cells in Tumor Microenvironment via IL-17 Pathway in Murine Hepatocarcinoma Model

    doi: 10.1371/journal.pone.0008922

    Figure Lengend Snippet: IL-9 strengthens the survival and protumor effect of mast cells in tumor microenvironment. (A and B) Mast cells upregulated the expressions of IL-9 by Treg cells. BMMCs were injected into tumor-bearing mice with IL-17 antibody or control antibody. Seven days later, the tumor-infiltrating Treg cells were isolated for the analysis of IL-9 by real time RT-PCR (A). Or the isolated Treg cells were cultured for 48 hours. The supernatant was used for IL-9 ELISA assay (B). (C and D) The interference of IL-9 signaling affected mast cell-mediated MDSC infiltration and mast cell-promoted tumor growth. 5×10 6 BMMCs were injected into tumor-bearing mice (n = 6) by i.v. injection. IL-9 neutralizing antibody or CD25 depleting antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 12, the tumor-infiltrating lymphocytes were used to analyze Gr-1 + CD11b + MDSCs by flow cytometry (C), and the tumor growth was monitored by measuring the length (L) and width (W) of tumors. The volume (V) of the tumor was calculated by the formula V = (L×W2)/2 (D). (E) IL-9 affected the survival of mast cells in tumor microenvironment. 1×10 6 CFSE-labeled BMMCs were directly injected into tumor tissue with multiple injection sites. IL-9 neutralizing antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. The tumor tissues were surgically excised on day 7 for fluorescent analysis of frozen sections.

    Article Snippet: 12 days later, 5×106 BMMCs were injected into tumor-bearing mice via tail vein, When indicated, the mice received i.p. injection of 100 µg of goat-anti-mouse IL-17 neutralizing antibody (R & D Systems, Minneapolis, MN) or isotype control 24 h and 1 h before BMMCs injection.

    Techniques: Injection, Mouse Assay, Isolation, Quantitative RT-PCR, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cytometry, Labeling

    Mast cell-induced IL-17 enhances suppressor function of Treg cells via upregulating CD39 and CD73. (A) The interference of IL-17 impaired the effect of mast cells on the suppressive function of Treg cells. 5×10 6 BMMCs were injected into tumor-bearing mice by i.v. injection. IL-17 or CCL2 neutralizing antibody or Gr-1 depleting antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 7, tumor-infiltrating Treg cells were isolated for suppression assay. (B and C) Mast cells had no effect on Treg cells expressing CTLA-4, IL-10 or TGF-β. BMMCs were injected into tumor-bearing mice. Seven days later, the tumor-infiltrating lymphocytes were isolated for the analysis of CTLA-4 by flow cytometry. The data showed the gated CD3 + Foxp3 + cells. (B) or IL-10 and TGF-β by real time RT-PCR (C). (D) Mast cells upregulated the expressions of CD39 and CD73 by Treg cells. BMMCs were injected into tumor-bearing mice with IL-17 antibody or control antibody. Seven days later, the tumor-infiltrating Treg cells were isolated for the analysis of CD39 and CD73 by real time RT-PCR and western blot. (E) IL-17 had no direct effect on Treg cells. IL-17 (20 ng/ml) was added to the cultured Treg cells for 12 hours. The cells were collected for the analysis of CD39 and CD73 by real time RT-PCR. F , Blockade of adenosine signaling pathway impaired mast cell-enhanced Treg cell function. BMMCs were injected into tumor-bearing mice. BM cells were used as control. Seven days later, the tumor-infiltrating Treg cells were isolated for suppression assay in the presence or absence of adenosine receptor A 2A antagonist SCH-58261 (100 ng/ml). The data shown were the representative of 2 independent experiments in which the similar results were obtained.

    Journal: PLoS ONE

    Article Title: Mast Cells Mobilize Myeloid-Derived Suppressor Cells and Treg Cells in Tumor Microenvironment via IL-17 Pathway in Murine Hepatocarcinoma Model

    doi: 10.1371/journal.pone.0008922

    Figure Lengend Snippet: Mast cell-induced IL-17 enhances suppressor function of Treg cells via upregulating CD39 and CD73. (A) The interference of IL-17 impaired the effect of mast cells on the suppressive function of Treg cells. 5×10 6 BMMCs were injected into tumor-bearing mice by i.v. injection. IL-17 or CCL2 neutralizing antibody or Gr-1 depleting antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 7, tumor-infiltrating Treg cells were isolated for suppression assay. (B and C) Mast cells had no effect on Treg cells expressing CTLA-4, IL-10 or TGF-β. BMMCs were injected into tumor-bearing mice. Seven days later, the tumor-infiltrating lymphocytes were isolated for the analysis of CTLA-4 by flow cytometry. The data showed the gated CD3 + Foxp3 + cells. (B) or IL-10 and TGF-β by real time RT-PCR (C). (D) Mast cells upregulated the expressions of CD39 and CD73 by Treg cells. BMMCs were injected into tumor-bearing mice with IL-17 antibody or control antibody. Seven days later, the tumor-infiltrating Treg cells were isolated for the analysis of CD39 and CD73 by real time RT-PCR and western blot. (E) IL-17 had no direct effect on Treg cells. IL-17 (20 ng/ml) was added to the cultured Treg cells for 12 hours. The cells were collected for the analysis of CD39 and CD73 by real time RT-PCR. F , Blockade of adenosine signaling pathway impaired mast cell-enhanced Treg cell function. BMMCs were injected into tumor-bearing mice. BM cells were used as control. Seven days later, the tumor-infiltrating Treg cells were isolated for suppression assay in the presence or absence of adenosine receptor A 2A antagonist SCH-58261 (100 ng/ml). The data shown were the representative of 2 independent experiments in which the similar results were obtained.

    Article Snippet: 12 days later, 5×106 BMMCs were injected into tumor-bearing mice via tail vein, When indicated, the mice received i.p. injection of 100 µg of goat-anti-mouse IL-17 neutralizing antibody (R & D Systems, Minneapolis, MN) or isotype control 24 h and 1 h before BMMCs injection.

    Techniques: Injection, Mouse Assay, Isolation, Suppression Assay, Expressing, Flow Cytometry, Cytometry, Quantitative RT-PCR, Western Blot, Cell Culture, Cell Function Assay

    Mast cells regulate MDSCs through IL-17 pathway. (A) Blockade of IL-17 prevented mast cell-mediated MDSC infiltration to tumor. 5×10 6 BMMCs were injected into tumor-bearing mice by i.v. injection. IL-17 neutralizing antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 7, the tumor-infiltrating lymphocytes were used to analyze Gr-1 + CD11b + MDSCs by flow cytometry (left). In addition, IL-17 neutralizing antibody was i.p. injected to the mice 24 h and 1 h before BMMCs injection. 2×10 6 CFSE-labeled MDSCs were injected into the mice two days later. The tumor tissues were surgically excised, and frozen sections were prepared and analyzed by fluorescence microscopy (right). (B) Blockade of IL-17 attenuated mast cell-mediated MDSC suppressive function. BMMCs were injected into tumor-bearing mice. IL-17 neutralizing antibody was i.p. injected to the mice at different time points. On day 7, tumor-infiltrating MDSCs were isolated for the suppression assay. (C) IL-17 was not expressed by H22 tumor cells, T cells or B cells. BMMCs were injected into tumor-bearing mice. Seven days later, tumor cells and TILs were isolated, respectively. The expression of IL-17 was analyzed by flow cytometry. (D and E) Mast cells upregulated the expression of IL-17 by MDSCs. Seven days after BMMCs injection, the isolated TILs were used for IL-17 expression analysis. The data showed the upregulation of IL-17 by CD11b + cells in BMMC group (D), and most of the gated IL-17 + cells expressed Gr-1 marker (E).

    Journal: PLoS ONE

    Article Title: Mast Cells Mobilize Myeloid-Derived Suppressor Cells and Treg Cells in Tumor Microenvironment via IL-17 Pathway in Murine Hepatocarcinoma Model

    doi: 10.1371/journal.pone.0008922

    Figure Lengend Snippet: Mast cells regulate MDSCs through IL-17 pathway. (A) Blockade of IL-17 prevented mast cell-mediated MDSC infiltration to tumor. 5×10 6 BMMCs were injected into tumor-bearing mice by i.v. injection. IL-17 neutralizing antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 7, the tumor-infiltrating lymphocytes were used to analyze Gr-1 + CD11b + MDSCs by flow cytometry (left). In addition, IL-17 neutralizing antibody was i.p. injected to the mice 24 h and 1 h before BMMCs injection. 2×10 6 CFSE-labeled MDSCs were injected into the mice two days later. The tumor tissues were surgically excised, and frozen sections were prepared and analyzed by fluorescence microscopy (right). (B) Blockade of IL-17 attenuated mast cell-mediated MDSC suppressive function. BMMCs were injected into tumor-bearing mice. IL-17 neutralizing antibody was i.p. injected to the mice at different time points. On day 7, tumor-infiltrating MDSCs were isolated for the suppression assay. (C) IL-17 was not expressed by H22 tumor cells, T cells or B cells. BMMCs were injected into tumor-bearing mice. Seven days later, tumor cells and TILs were isolated, respectively. The expression of IL-17 was analyzed by flow cytometry. (D and E) Mast cells upregulated the expression of IL-17 by MDSCs. Seven days after BMMCs injection, the isolated TILs were used for IL-17 expression analysis. The data showed the upregulation of IL-17 by CD11b + cells in BMMC group (D), and most of the gated IL-17 + cells expressed Gr-1 marker (E).

    Article Snippet: 12 days later, 5×106 BMMCs were injected into tumor-bearing mice via tail vein, When indicated, the mice received i.p. injection of 100 µg of goat-anti-mouse IL-17 neutralizing antibody (R & D Systems, Minneapolis, MN) or isotype control 24 h and 1 h before BMMCs injection.

    Techniques: Injection, Mouse Assay, Flow Cytometry, Cytometry, Labeling, Fluorescence, Microscopy, Isolation, Suppression Assay, Expressing, Marker

    Mast cell-induced IL-17 mediates Treg cell infiltration via upregulating chemokines CCL17 and CCL22. (A) The interference of IL-17 impaired the effect of mast cells on Treg cell infiltration. 5×10 6 BMMCs were injected into tumor-bearing mice by i.v. injection. IL-17 or CCL2 neutralizing antibody or Gr-1 depleting antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 7, the tumor-infiltrating lymphocytes were isolated to analyze CD3 + Foxp3 + Treg cells by flow cytometry. The results were combined from three mice. (B and C) Mast cells-induced IL-17 upregulated CCL17 and CCL22 expressions in tumor microenvironment. BMMCs were injected into tumor-bearing mice. IL-17 or Gr-1 antibody was i.p. injected to the mice at different time points. Seven days after BMMCs injection, the tumor tissues were used to analyze CCL17 and CCL22 expressions by real time RT-PCR (B) and ELISA (C). (D) The effect of CCL17 and CCL22 on Treg cell infiltration. BMMCs were injected into tumor-bearing mice. CCL17 or CCL22 neutralizing antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 7, the tumor-infiltrating lymphocytes were isolated to analyze CD3 + Foxp3 + Treg cells by flow cytometry. The results were combined from three mice.

    Journal: PLoS ONE

    Article Title: Mast Cells Mobilize Myeloid-Derived Suppressor Cells and Treg Cells in Tumor Microenvironment via IL-17 Pathway in Murine Hepatocarcinoma Model

    doi: 10.1371/journal.pone.0008922

    Figure Lengend Snippet: Mast cell-induced IL-17 mediates Treg cell infiltration via upregulating chemokines CCL17 and CCL22. (A) The interference of IL-17 impaired the effect of mast cells on Treg cell infiltration. 5×10 6 BMMCs were injected into tumor-bearing mice by i.v. injection. IL-17 or CCL2 neutralizing antibody or Gr-1 depleting antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 7, the tumor-infiltrating lymphocytes were isolated to analyze CD3 + Foxp3 + Treg cells by flow cytometry. The results were combined from three mice. (B and C) Mast cells-induced IL-17 upregulated CCL17 and CCL22 expressions in tumor microenvironment. BMMCs were injected into tumor-bearing mice. IL-17 or Gr-1 antibody was i.p. injected to the mice at different time points. Seven days after BMMCs injection, the tumor tissues were used to analyze CCL17 and CCL22 expressions by real time RT-PCR (B) and ELISA (C). (D) The effect of CCL17 and CCL22 on Treg cell infiltration. BMMCs were injected into tumor-bearing mice. CCL17 or CCL22 neutralizing antibody was i.p. injected to the mice 1 h, 2 days and 5 days after BMMCs injection. On day 7, the tumor-infiltrating lymphocytes were isolated to analyze CD3 + Foxp3 + Treg cells by flow cytometry. The results were combined from three mice.

    Article Snippet: 12 days later, 5×106 BMMCs were injected into tumor-bearing mice via tail vein, When indicated, the mice received i.p. injection of 100 µg of goat-anti-mouse IL-17 neutralizing antibody (R & D Systems, Minneapolis, MN) or isotype control 24 h and 1 h before BMMCs injection.

    Techniques: Injection, Mouse Assay, Isolation, Flow Cytometry, Cytometry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    A model of the closed loop among mast cells, MDSCs and Treg cells in tumor microenvironment. Under the guidance of SCF/c-kit signaling, mast cells migrate to and are activated in tumor microenvironment; the activated mast cells release a panel of factors, leading to CCL2 production and IL-17 upregulation in MDSCs; CCL2 signaling recruits more MDSCs, leading to more IL-17 production; IL-17 strengthens tumor inflammatory microenvironment, leading to the upregulation of IL-9, IL-10, IL-13, CCL17, CCL22, CD39 and CD73; IL-10 and IL-13 induce arginase 1 expression by MDSCs; CCL17 and CCL22 attract the migration of Treg cells; CD39 and CD73 enhance suppressor function of Treg cells; IL-9 produced by Treg cells maintains the survival of mast cells; MDSCs release active MMP9, through which soluble SCF is generated, thus further facilitating the migration and activation of mast cells.

    Journal: PLoS ONE

    Article Title: Mast Cells Mobilize Myeloid-Derived Suppressor Cells and Treg Cells in Tumor Microenvironment via IL-17 Pathway in Murine Hepatocarcinoma Model

    doi: 10.1371/journal.pone.0008922

    Figure Lengend Snippet: A model of the closed loop among mast cells, MDSCs and Treg cells in tumor microenvironment. Under the guidance of SCF/c-kit signaling, mast cells migrate to and are activated in tumor microenvironment; the activated mast cells release a panel of factors, leading to CCL2 production and IL-17 upregulation in MDSCs; CCL2 signaling recruits more MDSCs, leading to more IL-17 production; IL-17 strengthens tumor inflammatory microenvironment, leading to the upregulation of IL-9, IL-10, IL-13, CCL17, CCL22, CD39 and CD73; IL-10 and IL-13 induce arginase 1 expression by MDSCs; CCL17 and CCL22 attract the migration of Treg cells; CD39 and CD73 enhance suppressor function of Treg cells; IL-9 produced by Treg cells maintains the survival of mast cells; MDSCs release active MMP9, through which soluble SCF is generated, thus further facilitating the migration and activation of mast cells.

    Article Snippet: 12 days later, 5×106 BMMCs were injected into tumor-bearing mice via tail vein, When indicated, the mice received i.p. injection of 100 µg of goat-anti-mouse IL-17 neutralizing antibody (R & D Systems, Minneapolis, MN) or isotype control 24 h and 1 h before BMMCs injection.

    Techniques: Expressing, Migration, Produced, Generated, Activation Assay