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    Millipore il17a
    Production of mitochondrial O 2 • in human aortic endothelial cells (HAECs). (A) HAECs were treated with Ang II, <t>IL17A</t> and TNFα (1–100 ng/ml) or their combinations for 24 hours prior to measurements of mitochondrial O 2 • by
    Il17a, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 80 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il17a/product/Millipore
    Average 99 stars, based on 80 article reviews
    Price from $9.99 to $1999.99
    il17a - by Bioz Stars, 2020-05
    99/100 stars
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    99
    Millipore il 17 antibody
    ( A – H ) Expression and sources of <t>IL-17</t> in corneas of C57BL/6 and BALB/c mice. Relative mRNA levels of IL-17 ( A ) were significantly greater in the infected cornea of C57BL/6 versus BALB/c mice at 5 days after infection. No difference was detected
    Il 17 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il 17 antibody/product/Millipore
    Average 99 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    il 17 antibody - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    Image Search Results


    Production of mitochondrial O 2 • in human aortic endothelial cells (HAECs). (A) HAECs were treated with Ang II, IL17A and TNFα (1–100 ng/ml) or their combinations for 24 hours prior to measurements of mitochondrial O 2 • by

    Journal: Hypertension

    Article Title: Mitochondrial Cyclophilin D in Vascular Oxidative Stress and Hypertension

    doi: 10.1161/HYPERTENSIONAHA.115.07085

    Figure Lengend Snippet: Production of mitochondrial O 2 • in human aortic endothelial cells (HAECs). (A) HAECs were treated with Ang II, IL17A and TNFα (1–100 ng/ml) or their combinations for 24 hours prior to measurements of mitochondrial O 2 • by

    Article Snippet: Angiotensin II (Ang II), TNFα and IL17A were purchased from Sigma (St Luis, MO), Thermo Scientific (Rockford, IL) and Ebioscience (San Diego, CA).

    Techniques:

    Angiotensin II - cytokine interplay in hypertension. Blood pressure (A) and aortic mitochondrial O 2 • (B) in C57Bl/6J mice infused with IL17A (1.5 μg/day) and Etanercept (ETN, i.p. 0.2 mg/day). * P

    Journal: Hypertension

    Article Title: Mitochondrial Cyclophilin D in Vascular Oxidative Stress and Hypertension

    doi: 10.1161/HYPERTENSIONAHA.115.07085

    Figure Lengend Snippet: Angiotensin II - cytokine interplay in hypertension. Blood pressure (A) and aortic mitochondrial O 2 • (B) in C57Bl/6J mice infused with IL17A (1.5 μg/day) and Etanercept (ETN, i.p. 0.2 mg/day). * P

    Article Snippet: Angiotensin II (Ang II), TNFα and IL17A were purchased from Sigma (St Luis, MO), Thermo Scientific (Rockford, IL) and Ebioscience (San Diego, CA).

    Techniques: Mouse Assay

    Angiotensin II - cytokine interplay in vascular oxidative stress. in C57Bl/6J (WT) or Tg SOD2 mouse aorta Isolated three mm aortic segments were placed in RPMI tissue culture and treated ex vivo with Ang II (10 nM) or Ang II + IL17A (10 ng/ml) + TNFα

    Journal: Hypertension

    Article Title: Mitochondrial Cyclophilin D in Vascular Oxidative Stress and Hypertension

    doi: 10.1161/HYPERTENSIONAHA.115.07085

    Figure Lengend Snippet: Angiotensin II - cytokine interplay in vascular oxidative stress. in C57Bl/6J (WT) or Tg SOD2 mouse aorta Isolated three mm aortic segments were placed in RPMI tissue culture and treated ex vivo with Ang II (10 nM) or Ang II + IL17A (10 ng/ml) + TNFα

    Article Snippet: Angiotensin II (Ang II), TNFα and IL17A were purchased from Sigma (St Luis, MO), Thermo Scientific (Rockford, IL) and Ebioscience (San Diego, CA).

    Techniques: Isolation, Ex Vivo

    FGF2 cooperates with IL-17 to promote CIA pathogenesis. ( A ) Mean clinical scores and disease incidence of CIA in C57/BL6 wild type mice or Il17a −/− mice treated with empty virus (Ad-EV) or adenovirus expressing FGF2 (Ad–FGF2) once a week from day 15 after CIA induction (n = 10–14 per group). ( B ) H E histology of the representative ankle sections from C57/BL6 wild type or Il17a −/− mice treated as in ( A ). ( C ) Quantitative mRNA expression of KC, CXCL2, IL-6 and COX-2 in the joints of C57/BL6 wild type or Il17a −/− mice treated as in ( A ) (n = 3–4 per group). Data are representative of two ( A – C ) independent experiments (mean and s.e.m. in A , C ). * P

    Journal: Scientific Reports

    Article Title: FGF2 cooperates with IL-17 to promote autoimmune inflammation

    doi: 10.1038/s41598-017-07597-8

    Figure Lengend Snippet: FGF2 cooperates with IL-17 to promote CIA pathogenesis. ( A ) Mean clinical scores and disease incidence of CIA in C57/BL6 wild type mice or Il17a −/− mice treated with empty virus (Ad-EV) or adenovirus expressing FGF2 (Ad–FGF2) once a week from day 15 after CIA induction (n = 10–14 per group). ( B ) H E histology of the representative ankle sections from C57/BL6 wild type or Il17a −/− mice treated as in ( A ). ( C ) Quantitative mRNA expression of KC, CXCL2, IL-6 and COX-2 in the joints of C57/BL6 wild type or Il17a −/− mice treated as in ( A ) (n = 3–4 per group). Data are representative of two ( A – C ) independent experiments (mean and s.e.m. in A , C ). * P

    Article Snippet: Induction of CIA 8–10 weeks old Il17a −/− and its representative littermate control mice were injected intradermally with 100 μg emulsified CII (Sigma-Aldrich) plus CFA at several sites of the tail base on day 1 and day 21.

    Techniques: Mouse Assay, Expressing

    Arhgef12 is necessary for IL17A-induced RhoA activation and airway hypercontractility. ( A and B ) Western blot for phosphorylated myosin light chain in Arhgef12 –/– and WT mouse tracheal lysates and WT mouse tracheal lysates treated with or without Arhgef12 inhibitor Y16, with densitometry ( n = 4 mice per condition; veh, DMSO; * P

    Journal: JCI Insight

    Article Title: Arhgef12 drives IL17A-induced airway contractility and airway hyperresponsiveness in mice

    doi: 10.1172/jci.insight.123578

    Figure Lengend Snippet: Arhgef12 is necessary for IL17A-induced RhoA activation and airway hypercontractility. ( A and B ) Western blot for phosphorylated myosin light chain in Arhgef12 –/– and WT mouse tracheal lysates and WT mouse tracheal lysates treated with or without Arhgef12 inhibitor Y16, with densitometry ( n = 4 mice per condition; veh, DMSO; * P

    Article Snippet: Smooth muscle dissected from mouse trachea and treated overnight with murine IL17A (Peprotech) or early passage human airway smooth muscle cells (Lonza) treated overnight with human IL17A were homogenized in lysis buffer (MilliporeSigma RIPA buffer) with protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific).

    Techniques: Activation Assay, Western Blot, Mouse Assay

    LPS activates CD4+ T cells Lipopolysaccharide (LPS, 100 μg) or phosphate buffered saline (PBS) was administered to mice (Balb/c) intratracheally. BAL fluid and lungs were harvested at indicated days (D1 - D6). Lungs were digested as described in Methods. Levels of IL-17A, IL-17F, and IL-22 in BAL fluid were determined by a multiplexed immunoassay (A, B, C). Expression levels of CD69 in the CD3+CD4+ cell population in lung tissue were determined by flow cytometry; representative histograms of CD69 expression (shaded areas represent isotype control) (D), mean fluorescence intensity (MFI) for CD69 (E). Data are shown as geometrical mean ± SEM (n = 3 - 6 per group). *, LPS group vs respective PBS group (p

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

    Article Title: T cell pathways involving CTLA4 contribute to a model of acute lung injury 1

    doi: 10.4049/jimmunol.0903238

    Figure Lengend Snippet: LPS activates CD4+ T cells Lipopolysaccharide (LPS, 100 μg) or phosphate buffered saline (PBS) was administered to mice (Balb/c) intratracheally. BAL fluid and lungs were harvested at indicated days (D1 - D6). Lungs were digested as described in Methods. Levels of IL-17A, IL-17F, and IL-22 in BAL fluid were determined by a multiplexed immunoassay (A, B, C). Expression levels of CD69 in the CD3+CD4+ cell population in lung tissue were determined by flow cytometry; representative histograms of CD69 expression (shaded areas represent isotype control) (D), mean fluorescence intensity (MFI) for CD69 (E). Data are shown as geometrical mean ± SEM (n = 3 - 6 per group). *, LPS group vs respective PBS group (p

    Article Snippet: Levels of interleukin (IL)-6, TNF-α, IL-2, IL-4, IL-13, and IL-17A in BAL fluid were measured by a multiplexed immunoassay (Millipore, Billerica, MA), and IL-17F, IL-22, IL-23, and IL-27 were measured by a multiplexed immunoassay (Biolegend Inc., San Diego, CA).

    Techniques: Mouse Assay, Expressing, Flow Cytometry, Cytometry, Fluorescence

    αCTLA4 modifies LPS-induced cytokine production, CD69 and Foxp3 expression in CD4+ T cells Mice were injected intraperitoneally with anti-CTLA4 antibody (αCTLA4, 100 μg) or hamster IgG (IgG, 100 μg) one day prior to intratracheal administration of LPS (100 μg). D6 group was reinjected intraperitoneally with αCTLA4 (100 μg) on day 3. BAL fluid and lungs were collected at indicated days (D1 - D6) after the LPS exposure. Lungs were digested as described in Methods. Levels of IL-17A in BAL fluid were determined by a multiplexed immunoassay (A). Expression of CD69 and Foxp3 in the CD3+CD4+ cell population in lung tissue was determined by flow cytometry: representative histograms of CD69 and Foxp3 expression at day 2 (shaded areas represent isotype control) (B); mean fluorescence intensity (MFI) for CD69 and Foxp3 (C, D). Data are shown as geometric mean ± SEM (n = 3 - 6 per group). †, p

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

    Article Title: T cell pathways involving CTLA4 contribute to a model of acute lung injury 1

    doi: 10.4049/jimmunol.0903238

    Figure Lengend Snippet: αCTLA4 modifies LPS-induced cytokine production, CD69 and Foxp3 expression in CD4+ T cells Mice were injected intraperitoneally with anti-CTLA4 antibody (αCTLA4, 100 μg) or hamster IgG (IgG, 100 μg) one day prior to intratracheal administration of LPS (100 μg). D6 group was reinjected intraperitoneally with αCTLA4 (100 μg) on day 3. BAL fluid and lungs were collected at indicated days (D1 - D6) after the LPS exposure. Lungs were digested as described in Methods. Levels of IL-17A in BAL fluid were determined by a multiplexed immunoassay (A). Expression of CD69 and Foxp3 in the CD3+CD4+ cell population in lung tissue was determined by flow cytometry: representative histograms of CD69 and Foxp3 expression at day 2 (shaded areas represent isotype control) (B); mean fluorescence intensity (MFI) for CD69 and Foxp3 (C, D). Data are shown as geometric mean ± SEM (n = 3 - 6 per group). †, p

    Article Snippet: Levels of interleukin (IL)-6, TNF-α, IL-2, IL-4, IL-13, and IL-17A in BAL fluid were measured by a multiplexed immunoassay (Millipore, Billerica, MA), and IL-17F, IL-22, IL-23, and IL-27 were measured by a multiplexed immunoassay (Biolegend Inc., San Diego, CA).

    Techniques: Expressing, Mouse Assay, Injection, Flow Cytometry, Cytometry, Fluorescence

    Binding properties of the identified IL17A-specific fully human monoclonal antibodies. ( A ) ELISA-based comparison of IL17A binding of patient-derived antibodies 5M002, 5M007, 5M012, 5M024, benchmark antibody AIN457, and an irrelevant IgG1. Binding of

    Journal: mAbs

    Article Title: Mining the human autoantibody repertoire: Isolation of potent IL17A-neutralizing monoclonal antibodies from a patient with thymoma

    doi: 10.4161/mabs.36292

    Figure Lengend Snippet: Binding properties of the identified IL17A-specific fully human monoclonal antibodies. ( A ) ELISA-based comparison of IL17A binding of patient-derived antibodies 5M002, 5M007, 5M012, 5M024, benchmark antibody AIN457, and an irrelevant IgG1. Binding of

    Article Snippet: The IL17A binding ELISA was developed using ABTS (Sigma, A1888).

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay

    Anti-CD122 treatment prevents the conversion of Th17 cells to pathogenic Th1 cells in the pancreas. ( A ) The percentages of Th17 cells in pancreatic islets of 10-week-old NOD mice treated with control mAb or ChMBC7 for 7 weeks ( n = 7 in each group). ( B ) Schematic diagram showing experimental design of Th17 transfer experiments. ( C ) Representative FACS plots of IL-17A and IFN-γ expression in donor cells before and after the transfer. ( D and E ) IFN-γ and IL-17A production by donor CD4 + T cells from pancreatic islets ( D ) and spleen ( E ) in recipient mice treated with either control mAb or ChMBC7 ( n = 5 in each group). Data are representative of 3 ( A ) or 2 ( B – E ) independent experiments. Statistical data are mean ± SEM. P values are calculated using Student’s t test. * P

    Journal: JCI Insight

    Article Title: CD122 blockade restores immunological tolerance in autoimmune type 1 diabetes via multiple mechanisms

    doi: 10.1172/jci.insight.96600

    Figure Lengend Snippet: Anti-CD122 treatment prevents the conversion of Th17 cells to pathogenic Th1 cells in the pancreas. ( A ) The percentages of Th17 cells in pancreatic islets of 10-week-old NOD mice treated with control mAb or ChMBC7 for 7 weeks ( n = 7 in each group). ( B ) Schematic diagram showing experimental design of Th17 transfer experiments. ( C ) Representative FACS plots of IL-17A and IFN-γ expression in donor cells before and after the transfer. ( D and E ) IFN-γ and IL-17A production by donor CD4 + T cells from pancreatic islets ( D ) and spleen ( E ) in recipient mice treated with either control mAb or ChMBC7 ( n = 5 in each group). Data are representative of 3 ( A ) or 2 ( B – E ) independent experiments. Statistical data are mean ± SEM. P values are calculated using Student’s t test. * P

    Article Snippet: For intracellular IFN-γ and IL-17A detection, unfractionated cells were cultured in complete medium in the presence of phorbol myristate acetate (PMA, MilliporeSigma), ionomycin (MilliporeSigma), and brefeldin A (BioLegend) at 37°C for 4 hours.

    Techniques: Mouse Assay, FACS, Expressing

    Effect of digoxin on the balance of Th17 cells/Tregs. (A–D) Digoxin changed the percentage of Th17 cells and CD4 + forkhead box P3 (Foxp3) + Tregs among the mouse splenocytes. The concentrations of IL‐17A, IL‐10 and IL‐6

    Journal: British Journal of Pharmacology

    Article Title: Digoxin reduces atherosclerosis in apolipoprotein E‐deficient mice) Digoxin reduces atherosclerosis in apolipoprotein E‐deficient mice

    doi: 10.1111/bph.13453

    Figure Lengend Snippet: Effect of digoxin on the balance of Th17 cells/Tregs. (A–D) Digoxin changed the percentage of Th17 cells and CD4 + forkhead box P3 (Foxp3) + Tregs among the mouse splenocytes. The concentrations of IL‐17A, IL‐10 and IL‐6

    Article Snippet: The levels of mouse IL‐17A, IL‐10 and IL‐6 were measured using a mouse multi‐cytokine detection kit (Millipore, Billerica, MA, USA).

    Techniques:

    IL-1R signaling is required for host-protective T H 17 and T reg –T H 17 cell balance in vivo ( a ) Serial whole-body imaging of untreated Il17f thy1.1 mice, Il17f thy1.1 mice treated with depleting anti-Thy1.1 mAb or Il1r1 −/− mice or Il1r1 −/− mice treated with depleting anti-Thy1.1 mAb after inoculation with luminescent strain of C. rodentium and imaged at the indicated days post infection. ( b ) Colonization kinetic data from a represented as counts/sec at different time points post-infection with C. rodentium . ( c , d ) Quantitative ELISA of IL-17A ( c ) and IFNγ ( d ) in supernatants from cultured homogenates of colonic tissue collected from Il17f thy1.1 and Il1r1 −/− mice at the indicated times after inoculation with C. rodentium . ( e ) 3 × 10 6 of donor Il1r1 +/+ . Il17f thy1.1 (CD45.1/CD45.2) and Il1r1 −/− .Il17f thy1.1 (CD45.2) CD4 + T cells were mixed and co-transferred to recipient Tcrb −/− mice that were either uninfected or infected with C. rodentium 2 weeks post-reconstitution (see Supplementary Fig. 2d for schematic). Seven days later, expression of Thy1.1 (IL-17F) and intracellular Foxp3 by CD45.1 + and CD45.1 − splenic lymphocyte (SPL) and colonic lamina propria lymphocytes (LPL) from reconstituted recipient Tcrb −/− mice was analyzed (gated on activated CD4 + T cells). Numbers in each quadrant indicate the frequency of cells. Single plot represents equal frequencies of Il1r1 +/+ and Il1r1 −/− TCRβ + cells within reconstituted mice. ( f ) Frequencies of IL-17F (Thy1.1) cells in Il1r1 +/+ CD45.1 + and Il1r1 −/− CD45.1 − populations of SPL, MLN and LPL of uninfected and infected recipient Tcrb −/− mice. Data are: representative of one of two similar independent ( Il1r1 −/− mice treated with depleting anti-Thy1.1 mAb) or one of three similar independent experiments ( Il17f thy1.1 mice, Il17f thy1.1 mice treated with depleting anti-Thy1.1 mAb or Il1r1 −/− mice) a; pooled from two or three independent experiments with nine to eleven mice per group b; from two independent experiments with six mice per group c , d; representative of one of two similar independent experiments e; or pooled from two independent experiments with six mice per group f. Data are means and s.e.m. in b , c , d , f . N.S.= Not significant, * P

    Journal: Nature immunology

    Article Title: IL-1 signaling modulates STAT activation to antagonize retinoic acid signaling and control Th17–iTreg balance

    doi: 10.1038/ni.3099

    Figure Lengend Snippet: IL-1R signaling is required for host-protective T H 17 and T reg –T H 17 cell balance in vivo ( a ) Serial whole-body imaging of untreated Il17f thy1.1 mice, Il17f thy1.1 mice treated with depleting anti-Thy1.1 mAb or Il1r1 −/− mice or Il1r1 −/− mice treated with depleting anti-Thy1.1 mAb after inoculation with luminescent strain of C. rodentium and imaged at the indicated days post infection. ( b ) Colonization kinetic data from a represented as counts/sec at different time points post-infection with C. rodentium . ( c , d ) Quantitative ELISA of IL-17A ( c ) and IFNγ ( d ) in supernatants from cultured homogenates of colonic tissue collected from Il17f thy1.1 and Il1r1 −/− mice at the indicated times after inoculation with C. rodentium . ( e ) 3 × 10 6 of donor Il1r1 +/+ . Il17f thy1.1 (CD45.1/CD45.2) and Il1r1 −/− .Il17f thy1.1 (CD45.2) CD4 + T cells were mixed and co-transferred to recipient Tcrb −/− mice that were either uninfected or infected with C. rodentium 2 weeks post-reconstitution (see Supplementary Fig. 2d for schematic). Seven days later, expression of Thy1.1 (IL-17F) and intracellular Foxp3 by CD45.1 + and CD45.1 − splenic lymphocyte (SPL) and colonic lamina propria lymphocytes (LPL) from reconstituted recipient Tcrb −/− mice was analyzed (gated on activated CD4 + T cells). Numbers in each quadrant indicate the frequency of cells. Single plot represents equal frequencies of Il1r1 +/+ and Il1r1 −/− TCRβ + cells within reconstituted mice. ( f ) Frequencies of IL-17F (Thy1.1) cells in Il1r1 +/+ CD45.1 + and Il1r1 −/− CD45.1 − populations of SPL, MLN and LPL of uninfected and infected recipient Tcrb −/− mice. Data are: representative of one of two similar independent ( Il1r1 −/− mice treated with depleting anti-Thy1.1 mAb) or one of three similar independent experiments ( Il17f thy1.1 mice, Il17f thy1.1 mice treated with depleting anti-Thy1.1 mAb or Il1r1 −/− mice) a; pooled from two or three independent experiments with nine to eleven mice per group b; from two independent experiments with six mice per group c , d; representative of one of two similar independent experiments e; or pooled from two independent experiments with six mice per group f. Data are means and s.e.m. in b , c , d , f . N.S.= Not significant, * P

    Article Snippet: Only in cases where intracellular IL-17A was detected, cells were stimulated with PMA (50 ng/ml; Sigma) and ionomycin (750 ng/ml; Calbiochem) for 4 h in the presence of Golgi Plug (BD Pharmingen).

    Techniques: In Vivo, Imaging, Mouse Assay, Infection, Size-exclusion Chromatography, Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing

    IL-1β counteracts RA-driven IL-2/STAT5–dependent repression of T H 17 development ( a ) Expression of Thy1.1 (IL-17F) and Foxp3 from naïve OTII-Tg CD4 T cells (CD4 + CD25 − CD62L hi CD44 lo ) from OTII .Il17f thy1.1 mice activated with OVA peptide and IL-1β–deficient DCs for 4 days under T H 17 polarizing conditions, with or without IL-1β addition, and with or without addition of the indicated doses of rhIL-2 at day 4. ( b ) Frequencies of Foxp3 and IL-17F single-producers from OTII-Tg CD4 + T cells polarized as in a at indicated concentrations of rhIL-2 ( c ) Quantitative IL-2 ELISA from supernatants of naïve CD4 + T cells from Il17f thy1.1 mice that were activated with plate-bound anti-CD3 and soluble anti-CD28 under T H 17 polarizing conditions in absence (left) or presence (right) of IL-1β addition, with or without addition of addition of RA at the indicated concentrations (1–100 nM). ( d ) Expression of Cd25 and Cd122 transcripts by quantitative RT-PCR from naïve CD4 + T cells from Il17f thy1.1 mice that were activated with plate-bound anti-CD3 and soluble anti-CD28 under T H 17 polarizing conditions in absence or presence of IL-1β, with or without addition of RA (1 nM) at 60 h. Expression values are normalized to T H 0 controls. ( e ) Expression of IL-17A and Foxp3 from MACS-purified CD8 − CD4 + thymocytes isolated from WT B6 or Stat5 fl/fl . Cd4 -Cre mice that were activated with soluble anti-CD3 and Il1b −/− splenic DCs under T H 17 polarizing condition (IL-6+TGF-β with or without the indicated additions of RA (1 nM), anti-IL-2 (10 μg/ml) and IL-1β (20 ng/ml) for 4 days. ( f ) Frequencies of IL-17A + and Foxp3 + from CD4 + T cells from WT B6 or Stat5 fl/fl . Cd4 -Cre mice as treated in ( e ). Data are: representative of one of three similar independent experiments a ; pooled from three independent experiments with nine samples per group ( n = 9) b ; pooled from two independent experiments with six samples ( n = 6) c ; representative of two independent experiments with 6 samples per group ( n = 6) where individual data points represent each sample d ; representative of one of three similar independent experiments e ; or pooled from three independent experiments with ten samples ( n = 10) per group f. Data are means and s.e.m. in b , c , d , f ). ** P

    Journal: Nature immunology

    Article Title: IL-1 signaling modulates STAT activation to antagonize retinoic acid signaling and control Th17–iTreg balance

    doi: 10.1038/ni.3099

    Figure Lengend Snippet: IL-1β counteracts RA-driven IL-2/STAT5–dependent repression of T H 17 development ( a ) Expression of Thy1.1 (IL-17F) and Foxp3 from naïve OTII-Tg CD4 T cells (CD4 + CD25 − CD62L hi CD44 lo ) from OTII .Il17f thy1.1 mice activated with OVA peptide and IL-1β–deficient DCs for 4 days under T H 17 polarizing conditions, with or without IL-1β addition, and with or without addition of the indicated doses of rhIL-2 at day 4. ( b ) Frequencies of Foxp3 and IL-17F single-producers from OTII-Tg CD4 + T cells polarized as in a at indicated concentrations of rhIL-2 ( c ) Quantitative IL-2 ELISA from supernatants of naïve CD4 + T cells from Il17f thy1.1 mice that were activated with plate-bound anti-CD3 and soluble anti-CD28 under T H 17 polarizing conditions in absence (left) or presence (right) of IL-1β addition, with or without addition of addition of RA at the indicated concentrations (1–100 nM). ( d ) Expression of Cd25 and Cd122 transcripts by quantitative RT-PCR from naïve CD4 + T cells from Il17f thy1.1 mice that were activated with plate-bound anti-CD3 and soluble anti-CD28 under T H 17 polarizing conditions in absence or presence of IL-1β, with or without addition of RA (1 nM) at 60 h. Expression values are normalized to T H 0 controls. ( e ) Expression of IL-17A and Foxp3 from MACS-purified CD8 − CD4 + thymocytes isolated from WT B6 or Stat5 fl/fl . Cd4 -Cre mice that were activated with soluble anti-CD3 and Il1b −/− splenic DCs under T H 17 polarizing condition (IL-6+TGF-β with or without the indicated additions of RA (1 nM), anti-IL-2 (10 μg/ml) and IL-1β (20 ng/ml) for 4 days. ( f ) Frequencies of IL-17A + and Foxp3 + from CD4 + T cells from WT B6 or Stat5 fl/fl . Cd4 -Cre mice as treated in ( e ). Data are: representative of one of three similar independent experiments a ; pooled from three independent experiments with nine samples per group ( n = 9) b ; pooled from two independent experiments with six samples ( n = 6) c ; representative of two independent experiments with 6 samples per group ( n = 6) where individual data points represent each sample d ; representative of one of three similar independent experiments e ; or pooled from three independent experiments with ten samples ( n = 10) per group f. Data are means and s.e.m. in b , c , d , f ). ** P

    Article Snippet: Only in cases where intracellular IL-17A was detected, cells were stimulated with PMA (50 ng/ml; Sigma) and ionomycin (750 ng/ml; Calbiochem) for 4 h in the presence of Golgi Plug (BD Pharmingen).

    Techniques: Expressing, Mouse Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Magnetic Cell Separation, Purification, Isolation

    Deletion of SOCS3 abrogates the IL-1–dependent reversal of RA repression of T H 17 development ( a ) Expression of Foxp3 and IL-17A (right composite panel) in naïve CD4 + T cells from Socs3 fl/fl mice (control, red; Tat-Cre, blue) that were cultured under T H 17 polarizing conditions with the indicated additions of RA, IL-1β and IL-23 with or without Tat-Cre peptide and Td-Tomato protein expression (left single plot) from Rosa26 -floxed-STOP-TdTomato reporter mice ( Rosa26 TdTomato fl/+ ) treated with or without Tat-Cre peptide and cultured similarly under T H 17 conditions and analyzed for Cre-mediated deletion efficiency. Numbers indicate the frequencies in each quadrant. ( b ) Frequencies of IL-17A– and Foxp3–positive cells from CD4 + T cells treated as in ( a ). Data are representative of one of four (deletion efficiency analysis with Rosa26 TdTomato fl/+ ) or one of two ( Socs3 fl/fl with or without Tat-Cre) independent experiments ( a ) or are pooled from two independent experiments with 12 samples ( n = 12) per group ( b ). Data are means and s.e.m. in b . NS = not significant, ** P

    Journal: Nature immunology

    Article Title: IL-1 signaling modulates STAT activation to antagonize retinoic acid signaling and control Th17–iTreg balance

    doi: 10.1038/ni.3099

    Figure Lengend Snippet: Deletion of SOCS3 abrogates the IL-1–dependent reversal of RA repression of T H 17 development ( a ) Expression of Foxp3 and IL-17A (right composite panel) in naïve CD4 + T cells from Socs3 fl/fl mice (control, red; Tat-Cre, blue) that were cultured under T H 17 polarizing conditions with the indicated additions of RA, IL-1β and IL-23 with or without Tat-Cre peptide and Td-Tomato protein expression (left single plot) from Rosa26 -floxed-STOP-TdTomato reporter mice ( Rosa26 TdTomato fl/+ ) treated with or without Tat-Cre peptide and cultured similarly under T H 17 conditions and analyzed for Cre-mediated deletion efficiency. Numbers indicate the frequencies in each quadrant. ( b ) Frequencies of IL-17A– and Foxp3–positive cells from CD4 + T cells treated as in ( a ). Data are representative of one of four (deletion efficiency analysis with Rosa26 TdTomato fl/+ ) or one of two ( Socs3 fl/fl with or without Tat-Cre) independent experiments ( a ) or are pooled from two independent experiments with 12 samples ( n = 12) per group ( b ). Data are means and s.e.m. in b . NS = not significant, ** P

    Article Snippet: Only in cases where intracellular IL-17A was detected, cells were stimulated with PMA (50 ng/ml; Sigma) and ionomycin (750 ng/ml; Calbiochem) for 4 h in the presence of Golgi Plug (BD Pharmingen).

    Techniques: Expressing, Mouse Assay, Cell Culture

    IL-1β counteracts RA-dependent inhibition of T H 17 cell development ( a ) Naïve CD4 + T cells (CD4 + CD25 − CD62L hi CD44 lo ) from Il17f thy1.1 mice were activated with soluble anti-CD3 on DCs isolated from mesenteric lymph nodes (MLNs) of IL-1β–deficient ( Il1b −/− ) mice under T H 17 polarizing conditions, with the indicated additions of an at-RA inhibitor (LE540; 1 μM) or IL-1β (20 ng/ml). Cells recovered on day 4 were stained for surface CD4 and Thy1.1 (IL-17F), and intracellular IL-17A and Foxp3, and analyzed by flow cytometry. Numbers are percentages of cells in each quadrant. ( b ) Pooled data from a showing frequencies of IL-17A, IL-17F (Thy1.1) and Foxp3 single producers among CD4 + T cells. ( c ) Expression of Thy1.1 (IL-17F) and Foxp3 from naïve CD4 + T cells from Il17f thy1.1 mice activated with soluble anti-CD3 and Il1b −/− DCs under T H 17 polarizing conditions with or without IL-1β, in presence or absence of indicated concentration of at-RA at day 4. ( d ) Pooled data from c showing total frequencies of IL-17A–, IL-17F (Thy1.1)– and Foxp3–expressing CD4 + T cells. ( e ) Naïve CD4 + T cells from Il17f thy1.1 . Foxp3 gfp mice were activated with plate-bound anti-CD3 and soluble anti-CD28 under T H 17 polarizing conditions. On day 4 of primary culture the Thy1.1 − GFP + fraction of cells were sorted by flow cytometry and further cultured under T H 17 conditions with or without IL-1β (20 ng/ml), in the presence or absence of at-RA (1 nM). On day 4 of secondary culture, frequencies of IL-17F (Thy1.1) versus Foxp3-GFP + cells were determined after surface staining of Thy1.1 (IL-17F) and flow cytometry. Numbers are percentages of cells in the each quadrant. ( f ) Pooled data from secondary cultures from e showing frequencies of Foxp3 + (GFP) and IL-17F + (Thy1.1) CD4 + T cells. Data are representative of one of three similar independent experiments ( a ); pooled from three independent experiments with nine samples ( n = 9) per group ( b ); representative of one of three similar independent experiments ( c ); pooled from three experiments with twelve samples ( n = 12) per group ( d ); representative of one of two independent experiments ( e ); or pooled from two independent experiments with six samples ( n = 6) per group ( f ). Data are means and s.e.m. in b , d , f . ** P

    Journal: Nature immunology

    Article Title: IL-1 signaling modulates STAT activation to antagonize retinoic acid signaling and control Th17–iTreg balance

    doi: 10.1038/ni.3099

    Figure Lengend Snippet: IL-1β counteracts RA-dependent inhibition of T H 17 cell development ( a ) Naïve CD4 + T cells (CD4 + CD25 − CD62L hi CD44 lo ) from Il17f thy1.1 mice were activated with soluble anti-CD3 on DCs isolated from mesenteric lymph nodes (MLNs) of IL-1β–deficient ( Il1b −/− ) mice under T H 17 polarizing conditions, with the indicated additions of an at-RA inhibitor (LE540; 1 μM) or IL-1β (20 ng/ml). Cells recovered on day 4 were stained for surface CD4 and Thy1.1 (IL-17F), and intracellular IL-17A and Foxp3, and analyzed by flow cytometry. Numbers are percentages of cells in each quadrant. ( b ) Pooled data from a showing frequencies of IL-17A, IL-17F (Thy1.1) and Foxp3 single producers among CD4 + T cells. ( c ) Expression of Thy1.1 (IL-17F) and Foxp3 from naïve CD4 + T cells from Il17f thy1.1 mice activated with soluble anti-CD3 and Il1b −/− DCs under T H 17 polarizing conditions with or without IL-1β, in presence or absence of indicated concentration of at-RA at day 4. ( d ) Pooled data from c showing total frequencies of IL-17A–, IL-17F (Thy1.1)– and Foxp3–expressing CD4 + T cells. ( e ) Naïve CD4 + T cells from Il17f thy1.1 . Foxp3 gfp mice were activated with plate-bound anti-CD3 and soluble anti-CD28 under T H 17 polarizing conditions. On day 4 of primary culture the Thy1.1 − GFP + fraction of cells were sorted by flow cytometry and further cultured under T H 17 conditions with or without IL-1β (20 ng/ml), in the presence or absence of at-RA (1 nM). On day 4 of secondary culture, frequencies of IL-17F (Thy1.1) versus Foxp3-GFP + cells were determined after surface staining of Thy1.1 (IL-17F) and flow cytometry. Numbers are percentages of cells in the each quadrant. ( f ) Pooled data from secondary cultures from e showing frequencies of Foxp3 + (GFP) and IL-17F + (Thy1.1) CD4 + T cells. Data are representative of one of three similar independent experiments ( a ); pooled from three independent experiments with nine samples ( n = 9) per group ( b ); representative of one of three similar independent experiments ( c ); pooled from three experiments with twelve samples ( n = 12) per group ( d ); representative of one of two independent experiments ( e ); or pooled from two independent experiments with six samples ( n = 6) per group ( f ). Data are means and s.e.m. in b , d , f . ** P

    Article Snippet: Only in cases where intracellular IL-17A was detected, cells were stimulated with PMA (50 ng/ml; Sigma) and ionomycin (750 ng/ml; Calbiochem) for 4 h in the presence of Golgi Plug (BD Pharmingen).

    Techniques: Inhibition, Mouse Assay, Isolation, Staining, Flow Cytometry, Cytometry, Expressing, Concentration Assay, Cell Culture

    IL-17A-stimulated OPCs do not undergo apoptosis. OPCs from C57BL/6 and IL-17RA −/− pups were stimulated with 0, 1, or 100 ng/mL recombinant IL-17A for two days. ( A ) Total cells collected from each plate and supernatant were compared by

    Journal: Glia

    Article Title: IL-17A Activates ERK1/2 and Enhances Differentiation of Oligodendrocyte Progenitor Cells

    doi: 10.1002/glia.22783

    Figure Lengend Snippet: IL-17A-stimulated OPCs do not undergo apoptosis. OPCs from C57BL/6 and IL-17RA −/− pups were stimulated with 0, 1, or 100 ng/mL recombinant IL-17A for two days. ( A ) Total cells collected from each plate and supernatant were compared by

    Article Snippet: 0.1, 1, 10, or 100 ng/mL recombinant IL-17A, T3 (40 ng/mL, Sigma), or the same volume of medium was added 2 h later.

    Techniques: Recombinant

    IL-17A-stimulated OPCs exit the cell cycle. OPCs from C57BL/6 pups were cultured with 0, 0.01, 0.1, 1, 10, 100 ng/mL recombinant IL-17A for two days and mitotic cell division was measured by Ki67 staining ( A : Ki67, green; nuclei, blue) and blindly quantified

    Journal: Glia

    Article Title: IL-17A Activates ERK1/2 and Enhances Differentiation of Oligodendrocyte Progenitor Cells

    doi: 10.1002/glia.22783

    Figure Lengend Snippet: IL-17A-stimulated OPCs exit the cell cycle. OPCs from C57BL/6 pups were cultured with 0, 0.01, 0.1, 1, 10, 100 ng/mL recombinant IL-17A for two days and mitotic cell division was measured by Ki67 staining ( A : Ki67, green; nuclei, blue) and blindly quantified

    Article Snippet: 0.1, 1, 10, or 100 ng/mL recombinant IL-17A, T3 (40 ng/mL, Sigma), or the same volume of medium was added 2 h later.

    Techniques: Cell Culture, Recombinant, Staining

    ERK1/2 MAPK is activated in IL-17A-stimulated OPCs. OPCs from C57BL/6 and IL-17RA −/− pups were stimulated with 0, 1, or 100 ng/mL recombinant IL-17A for two days and protein for total ERK ( A ), p38 ( B ), and NFkB (p65) ( C ) were quantified

    Journal: Glia

    Article Title: IL-17A Activates ERK1/2 and Enhances Differentiation of Oligodendrocyte Progenitor Cells

    doi: 10.1002/glia.22783

    Figure Lengend Snippet: ERK1/2 MAPK is activated in IL-17A-stimulated OPCs. OPCs from C57BL/6 and IL-17RA −/− pups were stimulated with 0, 1, or 100 ng/mL recombinant IL-17A for two days and protein for total ERK ( A ), p38 ( B ), and NFkB (p65) ( C ) were quantified

    Article Snippet: 0.1, 1, 10, or 100 ng/mL recombinant IL-17A, T3 (40 ng/mL, Sigma), or the same volume of medium was added 2 h later.

    Techniques: Recombinant

    IL-17A enhanced OPC differentiation. ( A–C ) OPCs from postnatal rats were cultured with varying concentrations of recombinant IL-17A in the absence ( A ) or presence ( B ) of PDGF, stained with MBP by immunocytochemistry, and expression for each group

    Journal: Glia

    Article Title: IL-17A Activates ERK1/2 and Enhances Differentiation of Oligodendrocyte Progenitor Cells

    doi: 10.1002/glia.22783

    Figure Lengend Snippet: IL-17A enhanced OPC differentiation. ( A–C ) OPCs from postnatal rats were cultured with varying concentrations of recombinant IL-17A in the absence ( A ) or presence ( B ) of PDGF, stained with MBP by immunocytochemistry, and expression for each group

    Article Snippet: 0.1, 1, 10, or 100 ng/mL recombinant IL-17A, T3 (40 ng/mL, Sigma), or the same volume of medium was added 2 h later.

    Techniques: Cell Culture, Recombinant, Staining, Immunocytochemistry, Expressing

    IL-17A stimulated OPCs express a specific set of chemokines and cytokines. OPCs from C57BL/6 and IL-17RA −/− pups were stimulated with 0, 1, or 100 ng/mL recombinant IL-17A or 100 ng/mL IFN-γ for 2 days. Transcript levels for CNTF

    Journal: Glia

    Article Title: IL-17A Activates ERK1/2 and Enhances Differentiation of Oligodendrocyte Progenitor Cells

    doi: 10.1002/glia.22783

    Figure Lengend Snippet: IL-17A stimulated OPCs express a specific set of chemokines and cytokines. OPCs from C57BL/6 and IL-17RA −/− pups were stimulated with 0, 1, or 100 ng/mL recombinant IL-17A or 100 ng/mL IFN-γ for 2 days. Transcript levels for CNTF

    Article Snippet: 0.1, 1, 10, or 100 ng/mL recombinant IL-17A, T3 (40 ng/mL, Sigma), or the same volume of medium was added 2 h later.

    Techniques: Recombinant

    (a) Correlation between interleukin (IL)-17A concentrations in activated peripheral blood mononuclear cell (PBMCs) culture supernatants and type 2 diabetes mellitus (T2DM) duration. (b) Correlation between IL-17 + CD3 + CD8 − T cells in PBMCs and T2DM duration. (c) Correlation between IL-17A concentrations in activated PBMC culture supernatants and glycated haemoglobin (HbA1c) levels in patients with T2DM. (d) Correlation between IL-17 + CD3 + CD8 − T cells in PBMCs and HbA1c levels in patients with T2DM. (e) Correlation between IL-17A concentrations in activated PBMC culture supernatants and body mass index (BMI) in patients with T2DM. (f) Correlation between IL-17 + CD3 + CD8 − T cells in PBMCs and BMI in patients with T2DM. (g) Correlation between IL-17A concentrations in activated PBMC culture supernatants and IL-17 + CD3 + CD8 − T cells in PBMCs. Spearman correlation test was used ( P

    Journal: The Journal of International Medical Research

    Article Title: Th17 cell frequency and IL-17A concentrations in peripheral blood mononuclear cells and vitreous fluid from patients with diabetic retinopathy

    doi: 10.1177/0300060516672369

    Figure Lengend Snippet: (a) Correlation between interleukin (IL)-17A concentrations in activated peripheral blood mononuclear cell (PBMCs) culture supernatants and type 2 diabetes mellitus (T2DM) duration. (b) Correlation between IL-17 + CD3 + CD8 − T cells in PBMCs and T2DM duration. (c) Correlation between IL-17A concentrations in activated PBMC culture supernatants and glycated haemoglobin (HbA1c) levels in patients with T2DM. (d) Correlation between IL-17 + CD3 + CD8 − T cells in PBMCs and HbA1c levels in patients with T2DM. (e) Correlation between IL-17A concentrations in activated PBMC culture supernatants and body mass index (BMI) in patients with T2DM. (f) Correlation between IL-17 + CD3 + CD8 − T cells in PBMCs and BMI in patients with T2DM. (g) Correlation between IL-17A concentrations in activated PBMC culture supernatants and IL-17 + CD3 + CD8 − T cells in PBMCs. Spearman correlation test was used ( P

    Article Snippet: To study IL-17A production, PBMCs were stimulated with phytohaemagglutinin (5 µg/ml) at 2 × 106 cells/ml and 250 ng/ml ionomycin (Sigma-Aldrich, St Louis, MO, USA) for 48 h at 37℃ in a 5% CO2 atmosphere.

    Techniques:

    (a) Interleukin (IL)-17A levels in the vitreous fluid measured by enzyme-linked immunosorbent assay (ELISA). IL-17A levels were significantly higher in patients with DR than those in the control group (p

    Journal: The Journal of International Medical Research

    Article Title: Th17 cell frequency and IL-17A concentrations in peripheral blood mononuclear cells and vitreous fluid from patients with diabetic retinopathy

    doi: 10.1177/0300060516672369

    Figure Lengend Snippet: (a) Interleukin (IL)-17A levels in the vitreous fluid measured by enzyme-linked immunosorbent assay (ELISA). IL-17A levels were significantly higher in patients with DR than those in the control group (p

    Article Snippet: To study IL-17A production, PBMCs were stimulated with phytohaemagglutinin (5 µg/ml) at 2 × 106 cells/ml and 250 ng/ml ionomycin (Sigma-Aldrich, St Louis, MO, USA) for 48 h at 37℃ in a 5% CO2 atmosphere.

    Techniques: Enzyme-linked Immunosorbent Assay

    Interleukin (IL)-17A levels in stimulated cultured peripheral blood mononuclear cell (PBMC) supernatants measured by enzyme-linked immunosorbent assay (ELISA). (a) IL-17A levels in PBMC supernatants from patients with type 2 diabetes mellitus and no diabetic retinopathy (NDR) (n = 30), patients with diabetic retinopathy (DR) (n = 60) and healthy control subjects (n = 30). (b) IL-17A levels in PBMC supernatants from patients with nonproliferative DR (NPDR) (n = 30) and proliferative DR (PDR) (n = 30). PBMCs were cultured with phytohaemagglutinin (5 µg/ml) for 48 h. Data are represented as mean ± SD.

    Journal: The Journal of International Medical Research

    Article Title: Th17 cell frequency and IL-17A concentrations in peripheral blood mononuclear cells and vitreous fluid from patients with diabetic retinopathy

    doi: 10.1177/0300060516672369

    Figure Lengend Snippet: Interleukin (IL)-17A levels in stimulated cultured peripheral blood mononuclear cell (PBMC) supernatants measured by enzyme-linked immunosorbent assay (ELISA). (a) IL-17A levels in PBMC supernatants from patients with type 2 diabetes mellitus and no diabetic retinopathy (NDR) (n = 30), patients with diabetic retinopathy (DR) (n = 60) and healthy control subjects (n = 30). (b) IL-17A levels in PBMC supernatants from patients with nonproliferative DR (NPDR) (n = 30) and proliferative DR (PDR) (n = 30). PBMCs were cultured with phytohaemagglutinin (5 µg/ml) for 48 h. Data are represented as mean ± SD.

    Article Snippet: To study IL-17A production, PBMCs were stimulated with phytohaemagglutinin (5 µg/ml) at 2 × 106 cells/ml and 250 ng/ml ionomycin (Sigma-Aldrich, St Louis, MO, USA) for 48 h at 37℃ in a 5% CO2 atmosphere.

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

    Vγ4 + γδT17 development is blocked in the neonatal thymus in Sox13 mut/mut mice F 1 mice were backcrossed to Sox13 mut/mut (B6.SJL/NCI) mice to generate Sox13 mut/+ or Sox13 mut/mut neonates. ( a,c ) Intracellular IL-17A staining of thymocytes from day 0 and 5 neonates of the indicated type following stimulation with PMA+I, gated on Vγ4 + ( a ) or Vγ4 − ( c ) γδ T cells. ( b,d ) Quantification of Vγ4 + ( b ) or Vγ4 − ( d ) IL-17A + γδ T cells from day 0 and 5 neonatal thymi of the indicated type. Each symbol represents an individual mouse, horizontal and vertical bars represent the mean (± s.d.). * P ≤0.01, ** P ≤0.001, *** P ≤0.0001. Data are representative three experiments with at least 9 mice.

    Journal: Nature immunology

    Article Title: IL-17-committed V?4+ ?? T cell deficiency in a spontaneous Sox13 mutant CD45.1 congenic mouse substrain protects from dermatitis

    doi: 10.1038/ni.2585

    Figure Lengend Snippet: Vγ4 + γδT17 development is blocked in the neonatal thymus in Sox13 mut/mut mice F 1 mice were backcrossed to Sox13 mut/mut (B6.SJL/NCI) mice to generate Sox13 mut/+ or Sox13 mut/mut neonates. ( a,c ) Intracellular IL-17A staining of thymocytes from day 0 and 5 neonates of the indicated type following stimulation with PMA+I, gated on Vγ4 + ( a ) or Vγ4 − ( c ) γδ T cells. ( b,d ) Quantification of Vγ4 + ( b ) or Vγ4 − ( d ) IL-17A + γδ T cells from day 0 and 5 neonatal thymi of the indicated type. Each symbol represents an individual mouse, horizontal and vertical bars represent the mean (± s.d.). * P ≤0.01, ** P ≤0.001, *** P ≤0.0001. Data are representative three experiments with at least 9 mice.

    Article Snippet: For intracellular IL-17A staining, cells were stimulated for 2 hours with 50ng/ml phorbol myristate acetate (Sigma) and 1ug/ml ionomycin (EMD Biosciences) in brefeldin A (BD Biosciences).

    Techniques: Mouse Assay, Staining

    Vγ4 + γδT17 cells expand in draining LNs and home to inflamed ear skin ( a ) Intracellular IL-17A staining of PMA+I stimulated CLN cell suspensions from WT (B6/NCI) and Sox13 mut/mut (B6.SJL/NCI) mice treated with imiquimod or control cream for 5 days, gated on total γδ T cells. ( b,c ) Quantification of IL-17A + γδ T cells in CLNs ( b ) and blood ( c ) from mice treated as in ( a ). ( d ) Quantification Vγ4 + Vδ4 + γδ T cell number in CLNs (left panel) or frequency in ear skin (right panel, plotted as % of live cells) treated as in ( a ) for 3–7 days. ( e ) Quantification of donor T cells in the indicated tissues, plotted as percent of CD45.2 + donor cells, from day 2 imiquimod-treated Sox13 mut/mut recipients 3 hours after transfer of CLN cells from day 5 or 7 imiquimod-treated WT mice. ( f ) Quantification of donor T cells in the indicated tissues, plotted as percent of CD45.2 + (left panel) and IL-17A + CD45.2+ (right panel) donor cells from day 3 imiquimod-treated Sox13 mut/mut recipients 2 days after transfer of CLN cells from day 5 imiquimod-treated WT mice. ( g,h ) Quantification of Ly6G + CD11b + neutrophils, plotted as percent of CD45 + cells ( g ), and mRNA by RT-PCR ( h ) in ear skin from mice treated as in ( f ). Each symbol represents an individual mouse; horizontal bars represent the mean (± s.d.). * P ≤0.05, ** P ≤0.01, *** P ≤0.001. Data are representative of at least three experiments ( a–e,g ) or two experiments with at least 5 mice ( f,h ).

    Journal: Nature immunology

    Article Title: IL-17-committed V?4+ ?? T cell deficiency in a spontaneous Sox13 mutant CD45.1 congenic mouse substrain protects from dermatitis

    doi: 10.1038/ni.2585

    Figure Lengend Snippet: Vγ4 + γδT17 cells expand in draining LNs and home to inflamed ear skin ( a ) Intracellular IL-17A staining of PMA+I stimulated CLN cell suspensions from WT (B6/NCI) and Sox13 mut/mut (B6.SJL/NCI) mice treated with imiquimod or control cream for 5 days, gated on total γδ T cells. ( b,c ) Quantification of IL-17A + γδ T cells in CLNs ( b ) and blood ( c ) from mice treated as in ( a ). ( d ) Quantification Vγ4 + Vδ4 + γδ T cell number in CLNs (left panel) or frequency in ear skin (right panel, plotted as % of live cells) treated as in ( a ) for 3–7 days. ( e ) Quantification of donor T cells in the indicated tissues, plotted as percent of CD45.2 + donor cells, from day 2 imiquimod-treated Sox13 mut/mut recipients 3 hours after transfer of CLN cells from day 5 or 7 imiquimod-treated WT mice. ( f ) Quantification of donor T cells in the indicated tissues, plotted as percent of CD45.2 + (left panel) and IL-17A + CD45.2+ (right panel) donor cells from day 3 imiquimod-treated Sox13 mut/mut recipients 2 days after transfer of CLN cells from day 5 imiquimod-treated WT mice. ( g,h ) Quantification of Ly6G + CD11b + neutrophils, plotted as percent of CD45 + cells ( g ), and mRNA by RT-PCR ( h ) in ear skin from mice treated as in ( f ). Each symbol represents an individual mouse; horizontal bars represent the mean (± s.d.). * P ≤0.05, ** P ≤0.01, *** P ≤0.001. Data are representative of at least three experiments ( a–e,g ) or two experiments with at least 5 mice ( f,h ).

    Article Snippet: For intracellular IL-17A staining, cells were stimulated for 2 hours with 50ng/ml phorbol myristate acetate (Sigma) and 1ug/ml ionomycin (EMD Biosciences) in brefeldin A (BD Biosciences).

    Techniques: Staining, Mouse Assay, Reverse Transcription Polymerase Chain Reaction

    Sox13 mut/mut (B6.SJL/NCI) mice are protected from psoriasis-like dermatitis ( a ) Quantification of ear skin thickness, plotted as fraction increase relative to baseline (day 0), of WT (B6/NCI) and Sox13 mut/mut (B6.SJL/NCI) mice treated with imiquimod or control cream daily for 5 days. Boxes represent the mean (± s.d.). ( b ) H E staining of ear skin from WT and Sox13 mut/mut mice treated per ( a ) for 5 days. ( c ) Quantification of Ly6G + CD11b + neutrophils in ear skin cell suspensions from WT and Sox13 mut/mut mice treated per ( a ) for 3 or 5 days. Each symbol represents an individual mouse; horizontal bars represent the mean (± s.d.). ( d ) RT-PCR quantification of ear skin mRNA from WT and Sox13 mut/mut mice treated with control (−) or imiquimod cream for 3 or 5 days. Boxes represent the mean (± s.d.). ( e ) Intracellular IL-17A staining of ear skin cell suspensions from WT and Sox13 mut/mut mice treated as in ( a ) for 3 days and digested in the presence of Brefeldin A, gated on total γδ T cells. Mean (± s.d.) is indicated. ( f ) Transwell assay of neutrophil migration to ear skin supernatants prepared from WT and Sox13 mut/mut mice treated as in ( a ) for 3 days. Each symbol represents migration from an individual transwell, horizontal bars represent the mean (± s.d.). * P ≤0.05, ** P ≤0.01. Data are representative of three experiments with 3–6 mice ( a–d ), two experiments with 2–5 mice ( e ), and four experiments with 9 mice ( f ).

    Journal: Nature immunology

    Article Title: IL-17-committed V?4+ ?? T cell deficiency in a spontaneous Sox13 mutant CD45.1 congenic mouse substrain protects from dermatitis

    doi: 10.1038/ni.2585

    Figure Lengend Snippet: Sox13 mut/mut (B6.SJL/NCI) mice are protected from psoriasis-like dermatitis ( a ) Quantification of ear skin thickness, plotted as fraction increase relative to baseline (day 0), of WT (B6/NCI) and Sox13 mut/mut (B6.SJL/NCI) mice treated with imiquimod or control cream daily for 5 days. Boxes represent the mean (± s.d.). ( b ) H E staining of ear skin from WT and Sox13 mut/mut mice treated per ( a ) for 5 days. ( c ) Quantification of Ly6G + CD11b + neutrophils in ear skin cell suspensions from WT and Sox13 mut/mut mice treated per ( a ) for 3 or 5 days. Each symbol represents an individual mouse; horizontal bars represent the mean (± s.d.). ( d ) RT-PCR quantification of ear skin mRNA from WT and Sox13 mut/mut mice treated with control (−) or imiquimod cream for 3 or 5 days. Boxes represent the mean (± s.d.). ( e ) Intracellular IL-17A staining of ear skin cell suspensions from WT and Sox13 mut/mut mice treated as in ( a ) for 3 days and digested in the presence of Brefeldin A, gated on total γδ T cells. Mean (± s.d.) is indicated. ( f ) Transwell assay of neutrophil migration to ear skin supernatants prepared from WT and Sox13 mut/mut mice treated as in ( a ) for 3 days. Each symbol represents migration from an individual transwell, horizontal bars represent the mean (± s.d.). * P ≤0.05, ** P ≤0.01. Data are representative of three experiments with 3–6 mice ( a–d ), two experiments with 2–5 mice ( e ), and four experiments with 9 mice ( f ).

    Article Snippet: For intracellular IL-17A staining, cells were stimulated for 2 hours with 50ng/ml phorbol myristate acetate (Sigma) and 1ug/ml ionomycin (EMD Biosciences) in brefeldin A (BD Biosciences).

    Techniques: Mouse Assay, Staining, Reverse Transcription Polymerase Chain Reaction, Transwell Assay, Migration

    B6.SJL/NCI and B6.SJL/Tac mice lack Vγ4 + γδT17 cells ( a ) Flow cytometric detection of Vγ4 + CCR6 + γδ T cells in digested LN cell suspensions from B6/NCI and B6.SJL/NCI mice, gated on total γδ T cells. ( b ) Quantification of LN CCR6 + Vγ4 + and Vγ4 − γδ T cell frequency (plotted as % of total γδ T cells) and absolute number in B6/NCI and B6.SJL/NCI mice gated as in ( a ). ( c ) Intracellular IL-17A staining of digested LN cell suspensions from B6/NCI and B6.SJL/NCI mice following PMA+I stimulation, gated on total γδ T cells. ( d ) Quantification of the absolute number of LN IL-17 + Vγ4 + and Vγ4 − γδ T cells (left panel) and IL-17 + αβ T cells and TCR − cells (right panel) in B6/NCI and B6.SJL/NCI mice. ( e ) Quantification of the absolute number of SCART2 + , SCART2 − , and total CCR6 + γδ T cells in ear skin dermal cell suspensions from B6/NCI and B6.SJL/NCI mice. ( f ) Quantification of LN Vγ4 + CCR6 + γδ T cell frequency in B6.SJL and SJL mice from various vendors gated as in ( a ), plotted as % of total γδ T cells. Each symbol represents an individual mouse; horizontal and vertical bars represent the mean (± s.d.) ( b,d–f ). * P ≤0.01, ** P ≤0.005, *** P ≤0.001. Data are representative of three experiments with 6–7 mice ( a,b ), three experiments with 4–6 mice ( c,d ), five experiments with 9–11 mice ( e ), and at least two experiments with 4 mice for each strain ( f ).

    Journal: Nature immunology

    Article Title: IL-17-committed V?4+ ?? T cell deficiency in a spontaneous Sox13 mutant CD45.1 congenic mouse substrain protects from dermatitis

    doi: 10.1038/ni.2585

    Figure Lengend Snippet: B6.SJL/NCI and B6.SJL/Tac mice lack Vγ4 + γδT17 cells ( a ) Flow cytometric detection of Vγ4 + CCR6 + γδ T cells in digested LN cell suspensions from B6/NCI and B6.SJL/NCI mice, gated on total γδ T cells. ( b ) Quantification of LN CCR6 + Vγ4 + and Vγ4 − γδ T cell frequency (plotted as % of total γδ T cells) and absolute number in B6/NCI and B6.SJL/NCI mice gated as in ( a ). ( c ) Intracellular IL-17A staining of digested LN cell suspensions from B6/NCI and B6.SJL/NCI mice following PMA+I stimulation, gated on total γδ T cells. ( d ) Quantification of the absolute number of LN IL-17 + Vγ4 + and Vγ4 − γδ T cells (left panel) and IL-17 + αβ T cells and TCR − cells (right panel) in B6/NCI and B6.SJL/NCI mice. ( e ) Quantification of the absolute number of SCART2 + , SCART2 − , and total CCR6 + γδ T cells in ear skin dermal cell suspensions from B6/NCI and B6.SJL/NCI mice. ( f ) Quantification of LN Vγ4 + CCR6 + γδ T cell frequency in B6.SJL and SJL mice from various vendors gated as in ( a ), plotted as % of total γδ T cells. Each symbol represents an individual mouse; horizontal and vertical bars represent the mean (± s.d.) ( b,d–f ). * P ≤0.01, ** P ≤0.005, *** P ≤0.001. Data are representative of three experiments with 6–7 mice ( a,b ), three experiments with 4–6 mice ( c,d ), five experiments with 9–11 mice ( e ), and at least two experiments with 4 mice for each strain ( f ).

    Article Snippet: For intracellular IL-17A staining, cells were stimulated for 2 hours with 50ng/ml phorbol myristate acetate (Sigma) and 1ug/ml ionomycin (EMD Biosciences) in brefeldin A (BD Biosciences).

    Techniques: Mouse Assay, Flow Cytometry, Staining

    Reduction of the lesion size in the lungs of IL‐17A KO mice after M. tuberculosis infection. Wild‐type C57BL/6 or IL‐17A KO mice were inoculated i.t. with M. tuberculosis H37Rv. The mice were sacrificed 30, 60, and 120 days after infection, and formalin‐fixed sections were stained with hematoxylin and eosin. Representative lung tissue specimens from the wild‐type C57BL/6 mice (left panel) and the IL‐17A KO mice (right panel) are shown. Magnification, ×40 (A), ×400 (B).

    Journal: Immunity, Inflammation and Disease

    Article Title: Involvement of IL‐17A‐producing TCR γδ T cells in late protective immunity against pulmonary Mycobacterium tuberculosis infection

    doi: 10.1002/iid3.121

    Figure Lengend Snippet: Reduction of the lesion size in the lungs of IL‐17A KO mice after M. tuberculosis infection. Wild‐type C57BL/6 or IL‐17A KO mice were inoculated i.t. with M. tuberculosis H37Rv. The mice were sacrificed 30, 60, and 120 days after infection, and formalin‐fixed sections were stained with hematoxylin and eosin. Representative lung tissue specimens from the wild‐type C57BL/6 mice (left panel) and the IL‐17A KO mice (right panel) are shown. Magnification, ×40 (A), ×400 (B).

    Article Snippet: Flow cytometric analysis of intracellular cytokine To analyze the IL‐17A expression of the cells of the in vivo infection system, PIF cells or pulmonary lymphocytes from M. tuberculosis ‐infected mice were incubated with or without 1 μg/ml calcium ionophore A‐23187 (Calbiochem, San Diego, CA) and 25 ng/ml phorbol 12‐myristate 13‐acetate (PMA, Sigma) for 6 h at 37°C and 5% CO2 in the presence of GolgiPlug (BD).

    Techniques: Mouse Assay, Infection, Staining

    Th1‐type cytokine expression in the lungs of the surviving IL‐17A KO mice after pulmonary M. tuberculosis infection. Wild‐type C57BL/6 or IL‐17A KO mice were inoculated i.t. with M. tuberculosis H37Rv. The expression levels of IFN‐γ and TNF expression in the infected lungs were analyzed on day 250 after M. tuberculosis H37Rv infection by real‐time RT‐PCR (A), and production of IFN‐γ and TNF in the infected lungs was analyzed by the CBA system (B). The statistical analysis was performed with Student's t ‐test. Asterisks (*) indicate significant difference between two groups.

    Journal: Immunity, Inflammation and Disease

    Article Title: Involvement of IL‐17A‐producing TCR γδ T cells in late protective immunity against pulmonary Mycobacterium tuberculosis infection

    doi: 10.1002/iid3.121

    Figure Lengend Snippet: Th1‐type cytokine expression in the lungs of the surviving IL‐17A KO mice after pulmonary M. tuberculosis infection. Wild‐type C57BL/6 or IL‐17A KO mice were inoculated i.t. with M. tuberculosis H37Rv. The expression levels of IFN‐γ and TNF expression in the infected lungs were analyzed on day 250 after M. tuberculosis H37Rv infection by real‐time RT‐PCR (A), and production of IFN‐γ and TNF in the infected lungs was analyzed by the CBA system (B). The statistical analysis was performed with Student's t ‐test. Asterisks (*) indicate significant difference between two groups.

    Article Snippet: Flow cytometric analysis of intracellular cytokine To analyze the IL‐17A expression of the cells of the in vivo infection system, PIF cells or pulmonary lymphocytes from M. tuberculosis ‐infected mice were incubated with or without 1 μg/ml calcium ionophore A‐23187 (Calbiochem, San Diego, CA) and 25 ng/ml phorbol 12‐myristate 13‐acetate (PMA, Sigma) for 6 h at 37°C and 5% CO2 in the presence of GolgiPlug (BD).

    Techniques: Expressing, Mouse Assay, Infection, Quantitative RT-PCR, Crocin Bleaching Assay

    TCR γδ + T cells were the major IL‐17A‐producing cells in the lungs of M. tuberculosis ‐infected mice. Wild‐type C57BL/6 mice were inoculated i.t. with M. tuberculosis H37Rv or left untreated (A and B). The PIF cells (5 × 10 5 cells) were prepared on day 60, and were cultured with PPD (5 μg/ml) in the presence of naive spleen antigen‐presenting cells (1 × 10 5 cells) for 18 h at 37°C, and with GolgiPlug for the last 6 h. The cells were also stimulated with PMA and ionomycin. After the culture, the cells were surface stained with FITC‐CD3e, PerCP‐Cy5.5, or APC‐conjugated anti‐TCR Cβ and APC‐conjugated TCR Cδ mAbs. Surface‐stained cells were subjected to intercellular cytokine staining with a PE‐conjugated anti‐IL‐17A mAb. The samples were analyzed by FCM (A, naïve; B, infected). Wild‐type C57BL/6, IL‐17A KO, and TCR Cδ KO mice were inoculated i.t. with 1 × 10 3 CFU of M. tuberculosis H37Rv (C), and the CFU in the lungs was determined on day 60 after the infection. The statistical analysis was performed with ANOVA. Asterisks (*) indicate significant difference between two groups.

    Journal: Immunity, Inflammation and Disease

    Article Title: Involvement of IL‐17A‐producing TCR γδ T cells in late protective immunity against pulmonary Mycobacterium tuberculosis infection

    doi: 10.1002/iid3.121

    Figure Lengend Snippet: TCR γδ + T cells were the major IL‐17A‐producing cells in the lungs of M. tuberculosis ‐infected mice. Wild‐type C57BL/6 mice were inoculated i.t. with M. tuberculosis H37Rv or left untreated (A and B). The PIF cells (5 × 10 5 cells) were prepared on day 60, and were cultured with PPD (5 μg/ml) in the presence of naive spleen antigen‐presenting cells (1 × 10 5 cells) for 18 h at 37°C, and with GolgiPlug for the last 6 h. The cells were also stimulated with PMA and ionomycin. After the culture, the cells were surface stained with FITC‐CD3e, PerCP‐Cy5.5, or APC‐conjugated anti‐TCR Cβ and APC‐conjugated TCR Cδ mAbs. Surface‐stained cells were subjected to intercellular cytokine staining with a PE‐conjugated anti‐IL‐17A mAb. The samples were analyzed by FCM (A, naïve; B, infected). Wild‐type C57BL/6, IL‐17A KO, and TCR Cδ KO mice were inoculated i.t. with 1 × 10 3 CFU of M. tuberculosis H37Rv (C), and the CFU in the lungs was determined on day 60 after the infection. The statistical analysis was performed with ANOVA. Asterisks (*) indicate significant difference between two groups.

    Article Snippet: Flow cytometric analysis of intracellular cytokine To analyze the IL‐17A expression of the cells of the in vivo infection system, PIF cells or pulmonary lymphocytes from M. tuberculosis ‐infected mice were incubated with or without 1 μg/ml calcium ionophore A‐23187 (Calbiochem, San Diego, CA) and 25 ng/ml phorbol 12‐myristate 13‐acetate (PMA, Sigma) for 6 h at 37°C and 5% CO2 in the presence of GolgiPlug (BD).

    Techniques: Infection, Mouse Assay, Cell Culture, Staining

    BCG vaccination did not induce Th17 cells in the lungs after M. tuberculosis challenge. Wild‐type C57BL/6 mice were vaccinated with M. bovis BCG 30 or 60 days before M. tuberculosis H37Rv infection. The PIF cells (5 × 10 5 cells) were prepared on the 4th week after the M. tuberculosis infection, and were cultured with PPD (5 μg/ml) in the presence of naive spleen antigen presenting cells (1 × 10 5 cells) for 18 h at 37°C, and with GolgiPlug for the last 6 h. After the culture, the cells were surface stained with FITC‐CD3 and APC‐conjugated anti‐CD4. Surface‐stained cells were subjected to intercellular cytokine staining with a PE‐conjugated anti‐IL‐17A mAb. The samples were analyzed by FCM.

    Journal: Immunity, Inflammation and Disease

    Article Title: Involvement of IL‐17A‐producing TCR γδ T cells in late protective immunity against pulmonary Mycobacterium tuberculosis infection

    doi: 10.1002/iid3.121

    Figure Lengend Snippet: BCG vaccination did not induce Th17 cells in the lungs after M. tuberculosis challenge. Wild‐type C57BL/6 mice were vaccinated with M. bovis BCG 30 or 60 days before M. tuberculosis H37Rv infection. The PIF cells (5 × 10 5 cells) were prepared on the 4th week after the M. tuberculosis infection, and were cultured with PPD (5 μg/ml) in the presence of naive spleen antigen presenting cells (1 × 10 5 cells) for 18 h at 37°C, and with GolgiPlug for the last 6 h. After the culture, the cells were surface stained with FITC‐CD3 and APC‐conjugated anti‐CD4. Surface‐stained cells were subjected to intercellular cytokine staining with a PE‐conjugated anti‐IL‐17A mAb. The samples were analyzed by FCM.

    Article Snippet: Flow cytometric analysis of intracellular cytokine To analyze the IL‐17A expression of the cells of the in vivo infection system, PIF cells or pulmonary lymphocytes from M. tuberculosis ‐infected mice were incubated with or without 1 μg/ml calcium ionophore A‐23187 (Calbiochem, San Diego, CA) and 25 ng/ml phorbol 12‐myristate 13‐acetate (PMA, Sigma) for 6 h at 37°C and 5% CO2 in the presence of GolgiPlug (BD).

    Techniques: Mouse Assay, Infection, Cell Culture, Staining

    The survival of IL‐17A KO mice after mycobacterial infection. Groups of 11–13 mice were infected i.t. with 1 × 10 5 (A) or 1 × 10 3 (B) CFU of M. tuberculosis H37Rv, and the survival rates were monitored. The statistical significance of the differences in survival was determined by the generalized Wilcoxon's test. P = 0.0018 (A) and P = 0.0052 (B), respectively. The asterisk (*) indicates that the difference was considered to be significant.

    Journal: Immunity, Inflammation and Disease

    Article Title: Involvement of IL‐17A‐producing TCR γδ T cells in late protective immunity against pulmonary Mycobacterium tuberculosis infection

    doi: 10.1002/iid3.121

    Figure Lengend Snippet: The survival of IL‐17A KO mice after mycobacterial infection. Groups of 11–13 mice were infected i.t. with 1 × 10 5 (A) or 1 × 10 3 (B) CFU of M. tuberculosis H37Rv, and the survival rates were monitored. The statistical significance of the differences in survival was determined by the generalized Wilcoxon's test. P = 0.0018 (A) and P = 0.0052 (B), respectively. The asterisk (*) indicates that the difference was considered to be significant.

    Article Snippet: Flow cytometric analysis of intracellular cytokine To analyze the IL‐17A expression of the cells of the in vivo infection system, PIF cells or pulmonary lymphocytes from M. tuberculosis ‐infected mice were incubated with or without 1 μg/ml calcium ionophore A‐23187 (Calbiochem, San Diego, CA) and 25 ng/ml phorbol 12‐myristate 13‐acetate (PMA, Sigma) for 6 h at 37°C and 5% CO2 in the presence of GolgiPlug (BD).

    Techniques: Mouse Assay, Infection

    The Th1 immune response in the early phase against M. tuberculosis infection. Wild‐type C57BL/6 or IL‐17A KO mice were inoculated i.t. with M. tuberculosis H37Rv. Mice were sacrificed 30, 60, and 90 days after infection, and the lung lymphocytes were stimulated with PPD, and the percentages of IFN‐γ producing CD4 + cells in the CD3 + T cell population were determined using FCM (A). In some experiments, the concentrations of IFN‐γ in the supernatants of the lung lymphocytes were determined by an ELISA using the DuoSet ELISA development kit, according to the manufacturer's protocol (B). The statistical analysis was performed with Student's t ‐test. Asterisks (*) indicate significant difference between two groups.

    Journal: Immunity, Inflammation and Disease

    Article Title: Involvement of IL‐17A‐producing TCR γδ T cells in late protective immunity against pulmonary Mycobacterium tuberculosis infection

    doi: 10.1002/iid3.121

    Figure Lengend Snippet: The Th1 immune response in the early phase against M. tuberculosis infection. Wild‐type C57BL/6 or IL‐17A KO mice were inoculated i.t. with M. tuberculosis H37Rv. Mice were sacrificed 30, 60, and 90 days after infection, and the lung lymphocytes were stimulated with PPD, and the percentages of IFN‐γ producing CD4 + cells in the CD3 + T cell population were determined using FCM (A). In some experiments, the concentrations of IFN‐γ in the supernatants of the lung lymphocytes were determined by an ELISA using the DuoSet ELISA development kit, according to the manufacturer's protocol (B). The statistical analysis was performed with Student's t ‐test. Asterisks (*) indicate significant difference between two groups.

    Article Snippet: Flow cytometric analysis of intracellular cytokine To analyze the IL‐17A expression of the cells of the in vivo infection system, PIF cells or pulmonary lymphocytes from M. tuberculosis ‐infected mice were incubated with or without 1 μg/ml calcium ionophore A‐23187 (Calbiochem, San Diego, CA) and 25 ng/ml phorbol 12‐myristate 13‐acetate (PMA, Sigma) for 6 h at 37°C and 5% CO2 in the presence of GolgiPlug (BD).

    Techniques: Infection, Mouse Assay, Enzyme-linked Immunosorbent Assay

    The bacterial growth in various organs of IL‐17A KO mice after mycobacterial infection. Wild‐type C57BL/6 or IL‐17A KO mice were inoculated i.t. with 1 × 10 3 CFU of M. tuberculosis H37Rv, and the CFU in the lungs (A), livers (B), and spleens (C) was determined on days 30, 60, and 120 after the infection. The statistical analysis was performed with Student's t ‐test. The asterisk (*) indicates that there was a significant difference compared with wild‐type C57BL/6 mice.

    Journal: Immunity, Inflammation and Disease

    Article Title: Involvement of IL‐17A‐producing TCR γδ T cells in late protective immunity against pulmonary Mycobacterium tuberculosis infection

    doi: 10.1002/iid3.121

    Figure Lengend Snippet: The bacterial growth in various organs of IL‐17A KO mice after mycobacterial infection. Wild‐type C57BL/6 or IL‐17A KO mice were inoculated i.t. with 1 × 10 3 CFU of M. tuberculosis H37Rv, and the CFU in the lungs (A), livers (B), and spleens (C) was determined on days 30, 60, and 120 after the infection. The statistical analysis was performed with Student's t ‐test. The asterisk (*) indicates that there was a significant difference compared with wild‐type C57BL/6 mice.

    Article Snippet: Flow cytometric analysis of intracellular cytokine To analyze the IL‐17A expression of the cells of the in vivo infection system, PIF cells or pulmonary lymphocytes from M. tuberculosis ‐infected mice were incubated with or without 1 μg/ml calcium ionophore A‐23187 (Calbiochem, San Diego, CA) and 25 ng/ml phorbol 12‐myristate 13‐acetate (PMA, Sigma) for 6 h at 37°C and 5% CO2 in the presence of GolgiPlug (BD).

    Techniques: Mouse Assay, Infection

    Rapid changes in IL-17A and IL-6 serum cytokine concentrations in C57BL/6J mice following vector cannulations . Sera were prepared from blood collected from individual five-week old mice ( n = 4) randomly chosen one week prior to vector treatment (Day 0 on the graph). Mice were allowed to acclimate for seven days, followed by vector instillation of each salivary gland with 50 μl of vector solution containing 10 7 viral particles of either Ad5-LacZ or Ad5-IL17A vector. Sera were again prepared from blood collected from individual mice ( n = 11) at Day 5 and Day 12 post-treatment. Concentrations of cytokines were determined using the Luminex platform. To ensure sufficient quantities for testing, the sera of three individual mice of each experimental group were pooled. ND, not detected indicates levels below threshold detection.

    Journal: Arthritis Research & Therapy

    Article Title: Pathogenic effect of interleukin-17A in induction of Sj?gren's syndrome-like disease using adenovirus-mediated gene transfer

    doi: 10.1186/ar3207

    Figure Lengend Snippet: Rapid changes in IL-17A and IL-6 serum cytokine concentrations in C57BL/6J mice following vector cannulations . Sera were prepared from blood collected from individual five-week old mice ( n = 4) randomly chosen one week prior to vector treatment (Day 0 on the graph). Mice were allowed to acclimate for seven days, followed by vector instillation of each salivary gland with 50 μl of vector solution containing 10 7 viral particles of either Ad5-LacZ or Ad5-IL17A vector. Sera were again prepared from blood collected from individual mice ( n = 11) at Day 5 and Day 12 post-treatment. Concentrations of cytokines were determined using the Luminex platform. To ensure sufficient quantities for testing, the sera of three individual mice of each experimental group were pooled. ND, not detected indicates levels below threshold detection.

    Article Snippet: Determination of cytokines levels Measurements of IL-6 and IL-17A cytokine levels in sera samples were performed by an independent contractor (Millipore, Billerica, MA, USA) using Luminex® platform.

    Techniques: Mouse Assay, Plasmid Preparation, Luminex

    ( A – H ) Expression and sources of IL-17 in corneas of C57BL/6 and BALB/c mice. Relative mRNA levels of IL-17 ( A ) were significantly greater in the infected cornea of C57BL/6 versus BALB/c mice at 5 days after infection. No difference was detected

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Interleukin 17 Regulates Mer Tyrosine Kinase–Positive Cells in Pseudomonas aeruginosa Keratitis

    doi: 10.1167/iovs.14-14522

    Figure Lengend Snippet: ( A – H ) Expression and sources of IL-17 in corneas of C57BL/6 and BALB/c mice. Relative mRNA levels of IL-17 ( A ) were significantly greater in the infected cornea of C57BL/6 versus BALB/c mice at 5 days after infection. No difference was detected

    Article Snippet: Individual corneas from C57BL/6 and BALB/c mice, corneas from both strains after IL-17 antibody or IgG control treatment, and BALB/c corneas after IL-10 antibody or IgG treatment were harvested at 3 days after infection and incubated in 1 mL (1 mg/mL) type I collagenase (Sigma-Aldrich Corp.) in Hanks' balanced salt solution (Invitrogen) containing 5% fetal bovine serum (FBS) for 2 to 3 hours at 37°C.

    Techniques: Expressing, Mouse Assay, Infection

    ( A – G ) Interleukin 17 neutralizing antibody treatment of C57BL/6 mice. Similar clinical scores ( A ) were seen at 1, 3, and 5 days after infection in IL-17 antibody–treated compared with IgG-treated mice. Photographs taken with a slitlamp

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Interleukin 17 Regulates Mer Tyrosine Kinase–Positive Cells in Pseudomonas aeruginosa Keratitis

    doi: 10.1167/iovs.14-14522

    Figure Lengend Snippet: ( A – G ) Interleukin 17 neutralizing antibody treatment of C57BL/6 mice. Similar clinical scores ( A ) were seen at 1, 3, and 5 days after infection in IL-17 antibody–treated compared with IgG-treated mice. Photographs taken with a slitlamp

    Article Snippet: Individual corneas from C57BL/6 and BALB/c mice, corneas from both strains after IL-17 antibody or IgG control treatment, and BALB/c corneas after IL-10 antibody or IgG treatment were harvested at 3 days after infection and incubated in 1 mL (1 mg/mL) type I collagenase (Sigma-Aldrich Corp.) in Hanks' balanced salt solution (Invitrogen) containing 5% fetal bovine serum (FBS) for 2 to 3 hours at 37°C.

    Techniques: Mouse Assay, Infection

    ( A – C ) Effect of IL-17 neutralization on MerTK+ cells (and macrophages) in BALB/c and C57BL/6 mice and IL-10 neutralization in BALB/c mice. Increased MerTK+ cells and macrophages (F4/80+) were detected in IL-17 antibody over the IgG control treatment

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Interleukin 17 Regulates Mer Tyrosine Kinase–Positive Cells in Pseudomonas aeruginosa Keratitis

    doi: 10.1167/iovs.14-14522

    Figure Lengend Snippet: ( A – C ) Effect of IL-17 neutralization on MerTK+ cells (and macrophages) in BALB/c and C57BL/6 mice and IL-10 neutralization in BALB/c mice. Increased MerTK+ cells and macrophages (F4/80+) were detected in IL-17 antibody over the IgG control treatment

    Article Snippet: Individual corneas from C57BL/6 and BALB/c mice, corneas from both strains after IL-17 antibody or IgG control treatment, and BALB/c corneas after IL-10 antibody or IgG treatment were harvested at 3 days after infection and incubated in 1 mL (1 mg/mL) type I collagenase (Sigma-Aldrich Corp.) in Hanks' balanced salt solution (Invitrogen) containing 5% fetal bovine serum (FBS) for 2 to 3 hours at 37°C.

    Techniques: Neutralization, Mouse Assay

    ( A – G ) Effects of IL-17 neutralizing antibody treatment on TNF-α, MIP-2, IL-1β, and MPO. After IL-17 neutralization, mRNA levels of TNF-α ( A ), MIP-2 ( C ), and IL-1β ( E ) were significantly decreased in the infected

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Interleukin 17 Regulates Mer Tyrosine Kinase–Positive Cells in Pseudomonas aeruginosa Keratitis

    doi: 10.1167/iovs.14-14522

    Figure Lengend Snippet: ( A – G ) Effects of IL-17 neutralizing antibody treatment on TNF-α, MIP-2, IL-1β, and MPO. After IL-17 neutralization, mRNA levels of TNF-α ( A ), MIP-2 ( C ), and IL-1β ( E ) were significantly decreased in the infected

    Article Snippet: Individual corneas from C57BL/6 and BALB/c mice, corneas from both strains after IL-17 antibody or IgG control treatment, and BALB/c corneas after IL-10 antibody or IgG treatment were harvested at 3 days after infection and incubated in 1 mL (1 mg/mL) type I collagenase (Sigma-Aldrich Corp.) in Hanks' balanced salt solution (Invitrogen) containing 5% fetal bovine serum (FBS) for 2 to 3 hours at 37°C.

    Techniques: Neutralization, Infection

    ( A – H ) Interleukin 17 neutralizing antibody treatment of BALB/c mice. Clinical scores ( A ) were similar at 1 and 3 days after infection but were significantly higher in IL-17 antibody–treated compared with IgG-treated mice at 5 days after

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Interleukin 17 Regulates Mer Tyrosine Kinase–Positive Cells in Pseudomonas aeruginosa Keratitis

    doi: 10.1167/iovs.14-14522

    Figure Lengend Snippet: ( A – H ) Interleukin 17 neutralizing antibody treatment of BALB/c mice. Clinical scores ( A ) were similar at 1 and 3 days after infection but were significantly higher in IL-17 antibody–treated compared with IgG-treated mice at 5 days after

    Article Snippet: Individual corneas from C57BL/6 and BALB/c mice, corneas from both strains after IL-17 antibody or IgG control treatment, and BALB/c corneas after IL-10 antibody or IgG treatment were harvested at 3 days after infection and incubated in 1 mL (1 mg/mL) type I collagenase (Sigma-Aldrich Corp.) in Hanks' balanced salt solution (Invitrogen) containing 5% fetal bovine serum (FBS) for 2 to 3 hours at 37°C.

    Techniques: Mouse Assay, Infection