recombinant mouse il 27  (R&D Systems)

 
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    Name:
    Recombinant Mouse IL 27 NS0 expressed Protein CF
    Description:
    The Recombinant Mouse IL 27 NS0 expressed Protein from R D Systems is derived from NS0 The Recombinant Mouse IL 27 NS0 expressed Protein has been validated for the following applications Bioactivity
    Catalog Number:
    2799-ML-010/CF
    Price:
    329
    Category:
    Proteins and Enzymes
    Source:
    NS0-derived Recombinant Mouse IL-27 (NS0-expressed) Protein
    Applications:
    Bioactivity
    Purity:
    >97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie« Blue Staining.
    Conjugate:
    Unconjugated
    Size:
    10 ug
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    Structured Review

    R&D Systems recombinant mouse il 27
    Recombinant Mouse IL 27 NS0 expressed Protein CF
    The Recombinant Mouse IL 27 NS0 expressed Protein from R D Systems is derived from NS0 The Recombinant Mouse IL 27 NS0 expressed Protein has been validated for the following applications Bioactivity
    https://www.bioz.com/result/recombinant mouse il 27/product/R&D Systems
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant mouse il 27 - by Bioz Stars, 2021-05
    94/100 stars

    Images

    1) Product Images from "Interleukin 27 is a potential rescue therapy for acute severe colitis via interleukin-10 dependent, T cell independent attenuation of colonic mucosal innate immune responses"

    Article Title: Interleukin 27 is a potential rescue therapy for acute severe colitis via interleukin-10 dependent, T cell independent attenuation of colonic mucosal innate immune responses

    Journal: Inflammatory bowel diseases

    doi: 10.1097/MIB.0000000000001274

    IL-27 does not directly suppress macrophage CXCL2 expression To investigate the mechanism of IL-27 mediated immunosuppression, chemokine response in bone marrow derived macrophage (BMDM) ( a–c ), thioglycollate induced peritoneal macrophage (TPM) ( d e ) and colonic lamina propria F4/80 + macrophage ( f ) were assessed. ( a ) > 90% of M-CSF cultured bone marrow derived cells expressed F4/80 and IL-27Rα. ( b ) BMDM were stimulated for 5 hours with various concentrations of LPS as labelled +/− rIL27 (100ng/ml) and CXCL2 measured in supernatant by ELISA. Combined data from several experiments, generated from > 3 wells/experimental condition. LPS resulted in a robust CXCL2 response but there was no differential response in association with IL-27, t-test. ( c ) Exposure for > 48 hours prior to LPS stimulation did not impact CXCL2 response, t-test. ( d ) Peritoneal cell phenotype day 4 post IP thioglycollate injection. Cells expressed IL-27Rα. ( e ) Adherent cells were stimulated with 50ng/ml of LPS +/− 100ng/ml rIL27, for 5 or 72 hours. Again, LPS resulted in a robust CXCL2 response but there was no differential response in association with IL-27. ( f ) This was also seen in ex vivo colonic lamina propria F4/80+ cells stimulated for 24 hours with 50ng/ml of LPS +/− 100ng/ml rIL27, t-test. Results in C, E F presented from one experiment, with the same pattern of expression seen in repeat experiment. All data presented as mean + SEM.
    Figure Legend Snippet: IL-27 does not directly suppress macrophage CXCL2 expression To investigate the mechanism of IL-27 mediated immunosuppression, chemokine response in bone marrow derived macrophage (BMDM) ( a–c ), thioglycollate induced peritoneal macrophage (TPM) ( d e ) and colonic lamina propria F4/80 + macrophage ( f ) were assessed. ( a ) > 90% of M-CSF cultured bone marrow derived cells expressed F4/80 and IL-27Rα. ( b ) BMDM were stimulated for 5 hours with various concentrations of LPS as labelled +/− rIL27 (100ng/ml) and CXCL2 measured in supernatant by ELISA. Combined data from several experiments, generated from > 3 wells/experimental condition. LPS resulted in a robust CXCL2 response but there was no differential response in association with IL-27, t-test. ( c ) Exposure for > 48 hours prior to LPS stimulation did not impact CXCL2 response, t-test. ( d ) Peritoneal cell phenotype day 4 post IP thioglycollate injection. Cells expressed IL-27Rα. ( e ) Adherent cells were stimulated with 50ng/ml of LPS +/− 100ng/ml rIL27, for 5 or 72 hours. Again, LPS resulted in a robust CXCL2 response but there was no differential response in association with IL-27. ( f ) This was also seen in ex vivo colonic lamina propria F4/80+ cells stimulated for 24 hours with 50ng/ml of LPS +/− 100ng/ml rIL27, t-test. Results in C, E F presented from one experiment, with the same pattern of expression seen in repeat experiment. All data presented as mean + SEM.

    Techniques Used: Expressing, Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Generated, Injection, Ex Vivo

    IL-27 is associated with phosphorylated STAT-1 in colonic epithelial cells and significantly reduces pro-inflammatory cytokine profile in vivo ( a ) Immunohistochemistry for phosphorylated STAT-1 (Tyr701). Multifocal mucosal epithelial cells and scattered inflammatory cells within the lamina propria and submucosa show strong positive nuclear labeling for phosphorylated STAT-1 in response to oral LL-IL-27. Occasional cells show both strong nuclear and variable cytoplasmic labeling. There is no phosphorylated STAT-1 immunopositivity in colitic mice who received no treatment or treatment with LL-C. Diaminobenzidine chromogen and hematoxylin counterstain. This pattern of expression was seen in all mice per group (n=3). ( b ) Cytokine gene expression profile in distal colon homogenate was assessed by Taqman RT-PCR assays (Applied Biosystems) and analyzed with ddCt method of relative quantification; normalization to endogenous GAPDH control and level of expression compared to ethanol control group. Data, combined from 2 experiments, presented as mean + SEM. Il6 (p=0.003), Il1b (p=0.001) and Tnf (p=
    Figure Legend Snippet: IL-27 is associated with phosphorylated STAT-1 in colonic epithelial cells and significantly reduces pro-inflammatory cytokine profile in vivo ( a ) Immunohistochemistry for phosphorylated STAT-1 (Tyr701). Multifocal mucosal epithelial cells and scattered inflammatory cells within the lamina propria and submucosa show strong positive nuclear labeling for phosphorylated STAT-1 in response to oral LL-IL-27. Occasional cells show both strong nuclear and variable cytoplasmic labeling. There is no phosphorylated STAT-1 immunopositivity in colitic mice who received no treatment or treatment with LL-C. Diaminobenzidine chromogen and hematoxylin counterstain. This pattern of expression was seen in all mice per group (n=3). ( b ) Cytokine gene expression profile in distal colon homogenate was assessed by Taqman RT-PCR assays (Applied Biosystems) and analyzed with ddCt method of relative quantification; normalization to endogenous GAPDH control and level of expression compared to ethanol control group. Data, combined from 2 experiments, presented as mean + SEM. Il6 (p=0.003), Il1b (p=0.001) and Tnf (p=

    Techniques Used: In Vivo, Immunohistochemistry, Labeling, Mouse Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

    Oral delivery of IL-27 leads to improved histological colitis score and protection against ulceration in acute severe colitis ( a ) Cumulative histological colitis score based on 5 parameters (degree of inflammatory cell infiltrate/ mucosal ulceration/ crypt hyperplasia/ goblet cell depletion, presence of granuloma) with a maximum of 14. TNBS+LL-C vs. TNBS+ LL-IL-27, p=0.035. ( b ) Degree of distal colonic mucosa ulceration score, graded on a scale of 0–4. 0=none, 1=erosion, 2=mild ulceration, 3=moderate ulceration, 4=severe ulceration. TNBS+LL-C vs. TNBS+ LL-IL-27, p=0.008. Data generated from randomly selected animals from 2 experiments. n=5,5,6,7 for ethanol, TNBS, TNBS+LL-C and TNBS+LL-IL27, respectively. Mean + SEM. ( c ) Epithelial proliferation index was assessed by Ki67 immunohistochemistry on paraffin embedded distal colon sections. n=5/group from 2 experiments. Index derived from number of Ki67 positive cells/total number epithelial cells/ HPF, expressed as percentage. Mean + SEM % from 5 HPF per sample used in analysis. Colitis evoked a significant increase in proliferation index across all groups compared to ethanol control; p=0.009, 0.001 0.002 for TNBS alone, +LL-C and +LL-IL-27, respectively. There was no significant difference between TNBS alone vs. +LL-C vs. +LL-IL-27 (p=0.243), t-test. ( d ) Representative distal colon photomicrographs of each experimental group. There is full-thickness necrosis of the mucosa and marked submucosal inflammation and edema in both the TNBS alone and TNBS+LL-C treated mice. LL-IL-27 treatment significantly reduced mucosal necrosis and decreased inflammatory infiltrates. Slides were optically scanned with an Aperio AT2 digital slide scanner. Hematoxylin and eosin staining.
    Figure Legend Snippet: Oral delivery of IL-27 leads to improved histological colitis score and protection against ulceration in acute severe colitis ( a ) Cumulative histological colitis score based on 5 parameters (degree of inflammatory cell infiltrate/ mucosal ulceration/ crypt hyperplasia/ goblet cell depletion, presence of granuloma) with a maximum of 14. TNBS+LL-C vs. TNBS+ LL-IL-27, p=0.035. ( b ) Degree of distal colonic mucosa ulceration score, graded on a scale of 0–4. 0=none, 1=erosion, 2=mild ulceration, 3=moderate ulceration, 4=severe ulceration. TNBS+LL-C vs. TNBS+ LL-IL-27, p=0.008. Data generated from randomly selected animals from 2 experiments. n=5,5,6,7 for ethanol, TNBS, TNBS+LL-C and TNBS+LL-IL27, respectively. Mean + SEM. ( c ) Epithelial proliferation index was assessed by Ki67 immunohistochemistry on paraffin embedded distal colon sections. n=5/group from 2 experiments. Index derived from number of Ki67 positive cells/total number epithelial cells/ HPF, expressed as percentage. Mean + SEM % from 5 HPF per sample used in analysis. Colitis evoked a significant increase in proliferation index across all groups compared to ethanol control; p=0.009, 0.001 0.002 for TNBS alone, +LL-C and +LL-IL-27, respectively. There was no significant difference between TNBS alone vs. +LL-C vs. +LL-IL-27 (p=0.243), t-test. ( d ) Representative distal colon photomicrographs of each experimental group. There is full-thickness necrosis of the mucosa and marked submucosal inflammation and edema in both the TNBS alone and TNBS+LL-C treated mice. LL-IL-27 treatment significantly reduced mucosal necrosis and decreased inflammatory infiltrates. Slides were optically scanned with an Aperio AT2 digital slide scanner. Hematoxylin and eosin staining.

    Techniques Used: Generated, Immunohistochemistry, Derivative Assay, Mouse Assay, Staining

    An active inflammatory microenvironment is required for oral IL-27 to suppress inflammation in the colonic mucosa Pre-exposure to LL-IL-27 (10 oral gavages over 2 week period) prior to intra-rectal instillation of 2mg TNBS (SJL/J 6–8 week old male mice) did not impact acute colitis induction as measured by ( a ) disease activity index, ( b ) weight loss from starting weight, or ( c ) macroscopic colitis score (colon length and weight). Data presented combined from 2 separate experiments, n=10/group, as mean + SEM; ANOVA on Ranks Rank Sum test for pairwise comparisons.
    Figure Legend Snippet: An active inflammatory microenvironment is required for oral IL-27 to suppress inflammation in the colonic mucosa Pre-exposure to LL-IL-27 (10 oral gavages over 2 week period) prior to intra-rectal instillation of 2mg TNBS (SJL/J 6–8 week old male mice) did not impact acute colitis induction as measured by ( a ) disease activity index, ( b ) weight loss from starting weight, or ( c ) macroscopic colitis score (colon length and weight). Data presented combined from 2 separate experiments, n=10/group, as mean + SEM; ANOVA on Ranks Rank Sum test for pairwise comparisons.

    Techniques Used: Mouse Assay, Activity Assay

    Oral IL-27 reduces colonic neutrophil infiltrate associated with decreased CXC chemokine expression Distal colon inflammatory cell phenotype assessed by immunohistochemistry using ( a ) myeloperoxidase (MPO) and ( b ) F4/80 positivity to identify neutrophils and macrophages, respectively. Data generated from randomly selected animals from 2 experiments. n=5,5,6,6,8 for untreated, ethanol, TNBS, TNBS+LL-C and TNBS+LL-IL-27, respectively. Mean + SEM. ( a ) Treatment with IL-27 led to a significant reduction in MPO + neutrophil infiltration into the colonic mucosa (p=0.002, t-test). ( b ) F4/80 + macrophage infiltrate did not alter with LL-IL-27 treatment (p=0.250, t-test). ( c ) On day 2 post TNBS instillation, CXCL1 and CXCL2 but not CXCL5 protein (measured by ELISA and normalized to mg total protein concentration) were significantly reduced in distal colon homogenate in colitic mice treated with LL-IL-27 compared to LL-C (p=0.026, p=0.038 p=0.427 (t-test), respectively). Data represents mean + SEM, generated from 2 separate experiments, n=10 /group. ( d ) This IL-27 associated reduction in CXCL2 expression persisted as measured later in the experimental protocol (day 4 or time of death), p=0.001 and p=0.003, compared to TNBS plus LL-C and TNBS alone, respectively. Mean + SEM, generated from one experiment, n=3, 3, 8, 8, 11 for untreated, ethanol, TNBS, TNBS+LL-C and TNBS+LL-IL27, respectively. ( e ) Ex-vivo derived colitis associated lamina propria F4/80 + macrophage cells secreted CXCL2 measured by ELISA after 24 hours culture at 37°C (10 5 cells/200µl), and this was significantly reduced in cells exposed in vivo to LL-IL-27, p=0.001 (t-test). Data generated from 5 pooled distal colon samples per group.
    Figure Legend Snippet: Oral IL-27 reduces colonic neutrophil infiltrate associated with decreased CXC chemokine expression Distal colon inflammatory cell phenotype assessed by immunohistochemistry using ( a ) myeloperoxidase (MPO) and ( b ) F4/80 positivity to identify neutrophils and macrophages, respectively. Data generated from randomly selected animals from 2 experiments. n=5,5,6,6,8 for untreated, ethanol, TNBS, TNBS+LL-C and TNBS+LL-IL-27, respectively. Mean + SEM. ( a ) Treatment with IL-27 led to a significant reduction in MPO + neutrophil infiltration into the colonic mucosa (p=0.002, t-test). ( b ) F4/80 + macrophage infiltrate did not alter with LL-IL-27 treatment (p=0.250, t-test). ( c ) On day 2 post TNBS instillation, CXCL1 and CXCL2 but not CXCL5 protein (measured by ELISA and normalized to mg total protein concentration) were significantly reduced in distal colon homogenate in colitic mice treated with LL-IL-27 compared to LL-C (p=0.026, p=0.038 p=0.427 (t-test), respectively). Data represents mean + SEM, generated from 2 separate experiments, n=10 /group. ( d ) This IL-27 associated reduction in CXCL2 expression persisted as measured later in the experimental protocol (day 4 or time of death), p=0.001 and p=0.003, compared to TNBS plus LL-C and TNBS alone, respectively. Mean + SEM, generated from one experiment, n=3, 3, 8, 8, 11 for untreated, ethanol, TNBS, TNBS+LL-C and TNBS+LL-IL27, respectively. ( e ) Ex-vivo derived colitis associated lamina propria F4/80 + macrophage cells secreted CXCL2 measured by ELISA after 24 hours culture at 37°C (10 5 cells/200µl), and this was significantly reduced in cells exposed in vivo to LL-IL-27, p=0.001 (t-test). Data generated from 5 pooled distal colon samples per group.

    Techniques Used: Expressing, Immunohistochemistry, Generated, Enzyme-linked Immunosorbent Assay, Protein Concentration, Mouse Assay, T-Test, Ex Vivo, Derivative Assay, In Vivo

    IL-27 immunosuppression in acute TNBS colitis is T cell-independent and IL-10-dependent, evoking early IL-10 secretion in the colonic mucosa (a–d) 4mg TNBS instilled intra-rectally into Rag −/− mice. Data combined from 2 separate experiments. n=10/group. There was a significant difference in ( a ) DAI in mice treated with LL-IL-27 compared to LL-C on day 1 and 2, p=0.013 p=0.040 respectively, t-test, and ( b ) degree of weight loss from baseline on day 1 (p=0.019), then a trend just under significance on days 2 3 (p=0.06), t-test. ( c ) Distal colon homogenate CXCL2 protein was significantly reduced with LL-IL-27, p=0.028, ( d ) as was distal colon histology score, p=0.028, t-test. ( e–h ) 6mg TNBS instilled intra-rectally into IL-10 −/− mice. Data combined from 2 separate experiments. n=8/group. ( e ) DAI, p=0.237, 0.903, 0.681, respectively on day 1, 2 3, LL-IL-27 vs. LL-C, t-test, ( f ) weight loss from baseline, p=0.727, 0.751, 0.716, respectively on day 1, 2 and 3, LL-IL-27 vs. LL-C, t-test. ( g ) CXCL2 protein in distal colon homogenate, p=0.959 and ( h ) distal colon histology score, p=0.608, t-test, were not significantly different in mice treated with LL-IL-27 compared to LL-C. ( i ) On day 2 post TNBS instillation in SJL/J mice, there was a significant increase in distal colon IL-10 protein associated with IL-27, p=0.01, t-test. ( j ) Later in the treatment protocol, at day 4 or time of death, this difference in IL-10 protein expression was not apparent, p=0.653. At both time points, n=10/group, combined data from 2 experiments.
    Figure Legend Snippet: IL-27 immunosuppression in acute TNBS colitis is T cell-independent and IL-10-dependent, evoking early IL-10 secretion in the colonic mucosa (a–d) 4mg TNBS instilled intra-rectally into Rag −/− mice. Data combined from 2 separate experiments. n=10/group. There was a significant difference in ( a ) DAI in mice treated with LL-IL-27 compared to LL-C on day 1 and 2, p=0.013 p=0.040 respectively, t-test, and ( b ) degree of weight loss from baseline on day 1 (p=0.019), then a trend just under significance on days 2 3 (p=0.06), t-test. ( c ) Distal colon homogenate CXCL2 protein was significantly reduced with LL-IL-27, p=0.028, ( d ) as was distal colon histology score, p=0.028, t-test. ( e–h ) 6mg TNBS instilled intra-rectally into IL-10 −/− mice. Data combined from 2 separate experiments. n=8/group. ( e ) DAI, p=0.237, 0.903, 0.681, respectively on day 1, 2 3, LL-IL-27 vs. LL-C, t-test, ( f ) weight loss from baseline, p=0.727, 0.751, 0.716, respectively on day 1, 2 and 3, LL-IL-27 vs. LL-C, t-test. ( g ) CXCL2 protein in distal colon homogenate, p=0.959 and ( h ) distal colon histology score, p=0.608, t-test, were not significantly different in mice treated with LL-IL-27 compared to LL-C. ( i ) On day 2 post TNBS instillation in SJL/J mice, there was a significant increase in distal colon IL-10 protein associated with IL-27, p=0.01, t-test. ( j ) Later in the treatment protocol, at day 4 or time of death, this difference in IL-10 protein expression was not apparent, p=0.653. At both time points, n=10/group, combined data from 2 experiments.

    Techniques Used: Mouse Assay, Expressing

    Oral delivery of IL-27 suppresses acute severe colitis ( a ) LL-IL-27 evokes a decreased disease activity index (DAI) compared to LL-C; 5.8 vs. 8.2 on day 1, 5.3 vs. 8.9 on day 2 and 4.1 vs. 8.2, on day 3, maximum 12, *p=0.001. ( b ) LL-IL-27 led to significantly less weight loss than TNBS+LL-C or TNBS alone, p
    Figure Legend Snippet: Oral delivery of IL-27 suppresses acute severe colitis ( a ) LL-IL-27 evokes a decreased disease activity index (DAI) compared to LL-C; 5.8 vs. 8.2 on day 1, 5.3 vs. 8.9 on day 2 and 4.1 vs. 8.2, on day 3, maximum 12, *p=0.001. ( b ) LL-IL-27 led to significantly less weight loss than TNBS+LL-C or TNBS alone, p

    Techniques Used: Activity Assay

    2) Product Images from "A novel role for IL-27 in mediating the survival of activated mouse CD4 T lymphocytes: IL-27 inhibits activation induced T cell death"

    Article Title: A novel role for IL-27 in mediating the survival of activated mouse CD4 T lymphocytes: IL-27 inhibits activation induced T cell death

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

    doi: 10.4049/jimmunol.1201017

    IL-27 inhibits the activation of caspases. CFSE-labeled WT naïve CD4 + T cells were stimulated as in in the presence or absence of rIL-27. Anti-FasL blocking antibody was added where indicated either on day 0 (top row) or day 2 (bottom
    Figure Legend Snippet: IL-27 inhibits the activation of caspases. CFSE-labeled WT naïve CD4 + T cells were stimulated as in in the presence or absence of rIL-27. Anti-FasL blocking antibody was added where indicated either on day 0 (top row) or day 2 (bottom

    Techniques Used: Activation Assay, Labeling, Blocking Assay

    IL-27 is required for the maintenance of Treg population. (A-C) 4×10 5 CD45.1 + WT CD4 + CD45RB high cells were co-injected to Rag1 −/− mice with 1×10 5 CD45.2 CD4 + CD25 high cells sorted from WT (filled symbols) or IL-27R α
    Figure Legend Snippet: IL-27 is required for the maintenance of Treg population. (A-C) 4×10 5 CD45.1 + WT CD4 + CD45RB high cells were co-injected to Rag1 −/− mice with 1×10 5 CD45.2 CD4 + CD25 high cells sorted from WT (filled symbols) or IL-27R α

    Techniques Used: Injection, Mouse Assay

    IL-27 inhibits AICD through STAT3 signaling upstream of FasL-Fas. 1×10 5 CFSE-labeled naïve CD4 + T cells were stimulated with plate-bound anti-CD3ε and soluble anti-CD28 mAbs in the absence or presence of 10ng/ml rIL-27, as indicated
    Figure Legend Snippet: IL-27 inhibits AICD through STAT3 signaling upstream of FasL-Fas. 1×10 5 CFSE-labeled naïve CD4 + T cells were stimulated with plate-bound anti-CD3ε and soluble anti-CD28 mAbs in the absence or presence of 10ng/ml rIL-27, as indicated

    Techniques Used: Labeling

    IL-27 mediated enhancement of CD4 T cell survival involves a cell intrinsic mechanism. (A-C) 5×10 5 each of WT (CD45.1 + ) and IL-27R α −/− (CD45.1 − ) cells were co-injected intravenously to Rag1 −/− mice.
    Figure Legend Snippet: IL-27 mediated enhancement of CD4 T cell survival involves a cell intrinsic mechanism. (A-C) 5×10 5 each of WT (CD45.1 + ) and IL-27R α −/− (CD45.1 − ) cells were co-injected intravenously to Rag1 −/− mice.

    Techniques Used: Injection, Mouse Assay

    IL-27 increases cFLIP expression through a STAT3-mediated pathway. (A) Western blotting for detections of cFLIP protein. MACS sorted CD4 + CD25 − cells were stimulated as in previous figures, in the presence or absence of rIL-27. Cells were harvested
    Figure Legend Snippet: IL-27 increases cFLIP expression through a STAT3-mediated pathway. (A) Western blotting for detections of cFLIP protein. MACS sorted CD4 + CD25 − cells were stimulated as in previous figures, in the presence or absence of rIL-27. Cells were harvested

    Techniques Used: Expressing, Western Blot, Magnetic Cell Separation

    3) Product Images from "Thymic DCs derived IL-27 regulates the final maturation of CD4+ SP thymocytes"

    Article Title: Thymic DCs derived IL-27 regulates the final maturation of CD4+ SP thymocytes

    Journal: Scientific Reports

    doi: 10.1038/srep30448

    Deletion of p28-coding gene in DCs. ( A ) Mouse genotyping was performed using PCR. ( B ) p28 and Ebi3 mRNA levels were determined by quantitative PCR in total thymic DCs form CD11c-cre p28 flox/flox and WT mice. The IL-27 protein expression was measured by flow cytometry. ( C ) IL-27RA and gp130 mRNA expression was compared in CD4 SP thymocytes form CD11c-cre p28 flox/flox and WT mice. The experiments shown in B and C were repeated for more than three times and the data are presented as mean ± s.d. ** p
    Figure Legend Snippet: Deletion of p28-coding gene in DCs. ( A ) Mouse genotyping was performed using PCR. ( B ) p28 and Ebi3 mRNA levels were determined by quantitative PCR in total thymic DCs form CD11c-cre p28 flox/flox and WT mice. The IL-27 protein expression was measured by flow cytometry. ( C ) IL-27RA and gp130 mRNA expression was compared in CD4 SP thymocytes form CD11c-cre p28 flox/flox and WT mice. The experiments shown in B and C were repeated for more than three times and the data are presented as mean ± s.d. ** p

    Techniques Used: Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Mouse Assay, Expressing, Flow Cytometry, Cytometry

    The rescuing effect of intrathymically injected IL-27. CD11c-cre p28 flox/flox and WT mice received intrathymic injection of IL-27 or PBS. Two days later, the thymocytes were collected, stained with antibodies for CD4, CD8, CD69, Qa-2 and analyzed by flow cytometry. ( A ) Representative dot plots showing CD69 and Qa-2 expression in CD4 SP thymocytes. ( B ) The percentage of CD4 SP cells in total thymocytes, the percentage of SP4 cells in the CD4 SP population, and the absolute number of SP4 cells are shown. Each symbol represents an individual sample and the horizontal bar denotes the mean. * p
    Figure Legend Snippet: The rescuing effect of intrathymically injected IL-27. CD11c-cre p28 flox/flox and WT mice received intrathymic injection of IL-27 or PBS. Two days later, the thymocytes were collected, stained with antibodies for CD4, CD8, CD69, Qa-2 and analyzed by flow cytometry. ( A ) Representative dot plots showing CD69 and Qa-2 expression in CD4 SP thymocytes. ( B ) The percentage of CD4 SP cells in total thymocytes, the percentage of SP4 cells in the CD4 SP population, and the absolute number of SP4 cells are shown. Each symbol represents an individual sample and the horizontal bar denotes the mean. * p

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

    IL-27-enhanced development of Qa-2 + cells in CD4 + SP1 thymocyte cultures. ( A ) The profile of isolated CD4 + SP1 thymocytes was shown by the expression of 6C10 and CD69 (left), or by the expression of CD69 and Qa-2 (right). The purity of the sorted cells was 97–99%. ( B ) Freshly isolated CD4 + SP1 thymocytes from WT mice were cultured for 4 days in RPMI-1640 medium with or without the addition of IL-27, IL-30 or anti-IL-27p28. Acquisition of Qa-2 expression was detected by surface staining. Representative dot plots (left) and the percentage of Qa-2 + cells as well as the recovered live cells number (right) were presented. ( C ) SP1 cells were co-cultured with thymic DCs obtained from CD11c-cre p28 flox/flox or WT mice for 4 days in the presence or absence of IL-27. Qa-2 expression was evaluated by FACS analysis. Representative dot plots (left) and the percentage of Qa-2 + cells (right) are presented. The experiments were repeated for five times. * p
    Figure Legend Snippet: IL-27-enhanced development of Qa-2 + cells in CD4 + SP1 thymocyte cultures. ( A ) The profile of isolated CD4 + SP1 thymocytes was shown by the expression of 6C10 and CD69 (left), or by the expression of CD69 and Qa-2 (right). The purity of the sorted cells was 97–99%. ( B ) Freshly isolated CD4 + SP1 thymocytes from WT mice were cultured for 4 days in RPMI-1640 medium with or without the addition of IL-27, IL-30 or anti-IL-27p28. Acquisition of Qa-2 expression was detected by surface staining. Representative dot plots (left) and the percentage of Qa-2 + cells as well as the recovered live cells number (right) were presented. ( C ) SP1 cells were co-cultured with thymic DCs obtained from CD11c-cre p28 flox/flox or WT mice for 4 days in the presence or absence of IL-27. Qa-2 expression was evaluated by FACS analysis. Representative dot plots (left) and the percentage of Qa-2 + cells (right) are presented. The experiments were repeated for five times. * p

    Techniques Used: Isolation, Expressing, Mouse Assay, Cell Culture, Staining, FACS

    IL27- mediated signal in CD4 + SP thymocytes. ( A ) CD4 + SP thymocytes were isolated form CD11c-cre p28 flox/flox and WT mice, cultured in RPMI-1640 medium with IL-27. Cells were harvested at 0, 5, 15, 30 and 60 min after stimulation, Phosphorylated and total STAT1 and STAT3 were detected by Western blotting. The cropped gels have been run under the same experimental conditions. Representative blots are shown of three independent experiments. β-actin was used as a control. ( B ) SP3 cells from CD11c-cre p28 flox/flox mice and WT littermates were stimulated by 2 ng/ml IL-27 for 24 hours, cells were collected and the mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 were evaluated (n ≥ 3). The experiments were repeated for three times and the data are shown as mean ± s.d. ( C ) Total RNA was extracted from FACS-purified CD4 + SP thymocyte subsets from CD11c-cre p28 flox/flox mice and WT littermates. Quantitative PCR was performed to analyze mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 (n ≥ 3). * p
    Figure Legend Snippet: IL27- mediated signal in CD4 + SP thymocytes. ( A ) CD4 + SP thymocytes were isolated form CD11c-cre p28 flox/flox and WT mice, cultured in RPMI-1640 medium with IL-27. Cells were harvested at 0, 5, 15, 30 and 60 min after stimulation, Phosphorylated and total STAT1 and STAT3 were detected by Western blotting. The cropped gels have been run under the same experimental conditions. Representative blots are shown of three independent experiments. β-actin was used as a control. ( B ) SP3 cells from CD11c-cre p28 flox/flox mice and WT littermates were stimulated by 2 ng/ml IL-27 for 24 hours, cells were collected and the mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 were evaluated (n ≥ 3). The experiments were repeated for three times and the data are shown as mean ± s.d. ( C ) Total RNA was extracted from FACS-purified CD4 + SP thymocyte subsets from CD11c-cre p28 flox/flox mice and WT littermates. Quantitative PCR was performed to analyze mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 (n ≥ 3). * p

    Techniques Used: Isolation, Mouse Assay, Cell Culture, Western Blot, Expressing, FACS, Purification, Real-time Polymerase Chain Reaction

    IL-27 and its receptor expression in the thymus. ( A ) Total thymocyte, CD11c int CD45RA + pDC, CD11c hi CD8α + CD172α − and CD11c hi CD8α − CD172α + cDC, CD45 − Epcam hi Ly51 + cTEC, CD45 - Epcam hi Ly51 - mTEC and CD11b + F4/80 + macrophage were isolated from C57BL/6J mice. The expression of p28 and EBI3 mRNA was evaluated by quantitative PCR. nd, no detectable. ( B ) IL-27 protein expression in pDC, CD8α + and CD172α + cDC was analyzed by flow cytometry following intracellular staining with anti-IL-27. Representative histograms are shown on the left and the mean fluorescence intensity (MFI) on the right. ( C ) Purified DN, DP, CD8 SP and the four subsets of CD4 SP (SP1-4) thymocytes were analyzed for IL-27RA and gp130 mRNA expression using quantitative PCR. The lymph node (LN) naïve T cell was used as a positive control. All the experiments were repeated for more than three times and the data are shown as mean ± s.d. * p
    Figure Legend Snippet: IL-27 and its receptor expression in the thymus. ( A ) Total thymocyte, CD11c int CD45RA + pDC, CD11c hi CD8α + CD172α − and CD11c hi CD8α − CD172α + cDC, CD45 − Epcam hi Ly51 + cTEC, CD45 - Epcam hi Ly51 - mTEC and CD11b + F4/80 + macrophage were isolated from C57BL/6J mice. The expression of p28 and EBI3 mRNA was evaluated by quantitative PCR. nd, no detectable. ( B ) IL-27 protein expression in pDC, CD8α + and CD172α + cDC was analyzed by flow cytometry following intracellular staining with anti-IL-27. Representative histograms are shown on the left and the mean fluorescence intensity (MFI) on the right. ( C ) Purified DN, DP, CD8 SP and the four subsets of CD4 SP (SP1-4) thymocytes were analyzed for IL-27RA and gp130 mRNA expression using quantitative PCR. The lymph node (LN) naïve T cell was used as a positive control. All the experiments were repeated for more than three times and the data are shown as mean ± s.d. * p

    Techniques Used: Expressing, Isolation, Mouse Assay, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry, Staining, Fluorescence, Purification, Positive Control

    4) Product Images from "Interleukin 27 (IL-27) Alleviates Bone Loss in Estrogen-deficient Conditions by Induction of Early Growth Response-2 Gene *"

    Article Title: Interleukin 27 (IL-27) Alleviates Bone Loss in Estrogen-deficient Conditions by Induction of Early Growth Response-2 Gene *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M116.764779

    A, mRNA expression of IL-27 was determined in PBMCs of sham and Ovx animals. B, serum level of IL-27 in sham and Ovx mice. n = 8 mice/group. Data are presented mean ± S.E. and are representative of three independent experiments. ***, p
    Figure Legend Snippet: A, mRNA expression of IL-27 was determined in PBMCs of sham and Ovx animals. B, serum level of IL-27 in sham and Ovx mice. n = 8 mice/group. Data are presented mean ± S.E. and are representative of three independent experiments. ***, p

    Techniques Used: Expressing, Mouse Assay

    % CD4 + T cell populations in the BM of Sham, Sham + IL-27, Ovx, and Ovx + IL-27 groups were analyzed by flow cytometry. A, quantitative representation of the FACS data. B, % CD4 T cells in different groups. C, OPG/RANKL ratio was determined in B cells of Sham, Sham + IL-27, Ovx, Ovx + IL-27 groups. D, representative images B220 + cells of the FACS data. E, % B cells in BM of Sham, Sham + IL-27, Ovx, and Ovx + IL-27 groups was analyzed by flow cytometry. n = 8 mice/group; data are presented mean ± S.E. and are representative of three independent experiments. ***, p
    Figure Legend Snippet: % CD4 + T cell populations in the BM of Sham, Sham + IL-27, Ovx, and Ovx + IL-27 groups were analyzed by flow cytometry. A, quantitative representation of the FACS data. B, % CD4 T cells in different groups. C, OPG/RANKL ratio was determined in B cells of Sham, Sham + IL-27, Ovx, Ovx + IL-27 groups. D, representative images B220 + cells of the FACS data. E, % B cells in BM of Sham, Sham + IL-27, Ovx, and Ovx + IL-27 groups was analyzed by flow cytometry. n = 8 mice/group; data are presented mean ± S.E. and are representative of three independent experiments. ***, p

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

    Levels of IL-27 in healthy, osteopenic, and osteoporotic subjects and its correlation with BMD data. A, demographic characteristics of post-menopausal women divided into normal, osteopenic, and osteoporotic subjects based on BMD measurements. B, IL-27 levels in normal, osteopenic, and osteoporotic post-menopausal women. C, correlation of IL-27 serum levels and BMD values. D, mRNA expression of Egr-2 in PBMCs isolated from normal and osteoporotic subjects. Data are shown as mean ± S.E. for normal osteopenic and osteoporotic groups. NS , non-significant. ***, p
    Figure Legend Snippet: Levels of IL-27 in healthy, osteopenic, and osteoporotic subjects and its correlation with BMD data. A, demographic characteristics of post-menopausal women divided into normal, osteopenic, and osteoporotic subjects based on BMD measurements. B, IL-27 levels in normal, osteopenic, and osteoporotic post-menopausal women. C, correlation of IL-27 serum levels and BMD values. D, mRNA expression of Egr-2 in PBMCs isolated from normal and osteoporotic subjects. Data are shown as mean ± S.E. for normal osteopenic and osteoporotic groups. NS , non-significant. ***, p

    Techniques Used: Expressing, Isolation

    IL-27 initiates Egr-2 activation to induce Tr1 regulatory cell development and in vitro treatment of IL-27 induce IL-10 producing Tr1 cells. A, transcript levels of Egr-2 were determined in CD4 + T cells isolated from all groups. B, % of CD49b + LAG3 + Tr1 cells was measured by FACS. C, quantitative representation of the FACS data. D, CD4 + T cells were treated with IL-27 ( in vitro ) and percentage of CD49b + LAG3 + Tr1 cells were measured by FACS, and level of IL-10 cytokine was measured in gated Tr1 cells. E and F, quantitative representation of FACS data. Effect of IL-6 on Tr1 cells ( G and H ). n = 8 mice/group. Data are presented mean ± S.E. and are representative of three independent experiments. *, p
    Figure Legend Snippet: IL-27 initiates Egr-2 activation to induce Tr1 regulatory cell development and in vitro treatment of IL-27 induce IL-10 producing Tr1 cells. A, transcript levels of Egr-2 were determined in CD4 + T cells isolated from all groups. B, % of CD49b + LAG3 + Tr1 cells was measured by FACS. C, quantitative representation of the FACS data. D, CD4 + T cells were treated with IL-27 ( in vitro ) and percentage of CD49b + LAG3 + Tr1 cells were measured by FACS, and level of IL-10 cytokine was measured in gated Tr1 cells. E and F, quantitative representation of FACS data. Effect of IL-6 on Tr1 cells ( G and H ). n = 8 mice/group. Data are presented mean ± S.E. and are representative of three independent experiments. *, p

    Techniques Used: Activation Assay, In Vitro, Isolation, FACS, Mouse Assay

    Ovx mice treated with IL-27 prevented the loss of femoral bone microarchitecture. A, representative images of femoral bones of different groups. B, BV/TV; C, Tb.Th; D, Tb.N; E, Tb.Sp; F, SMI; and G, Tb.Pf. Statistical analysis was performed by ANOVA method followed by the Newman-Keuls test of significance using Prism version 3.0 software. n = 8 mice/group; data are presented as mean ± S.E.; ***, p
    Figure Legend Snippet: Ovx mice treated with IL-27 prevented the loss of femoral bone microarchitecture. A, representative images of femoral bones of different groups. B, BV/TV; C, Tb.Th; D, Tb.N; E, Tb.Sp; F, SMI; and G, Tb.Pf. Statistical analysis was performed by ANOVA method followed by the Newman-Keuls test of significance using Prism version 3.0 software. n = 8 mice/group; data are presented as mean ± S.E.; ***, p

    Techniques Used: Mouse Assay, Software

    IL-27 inhibits mouse calvarial osteoblast apoptosis by enhancing Egr-2 expression and suppressing inhibitory effect of CD4 + T cells isolated from Ovx mice on osteoblast differentiation. A, representative images of apoptosis assay done by annexin and PI staining. B, percentage of apoptotic cells in each set. Transcript levels of Egr-2 ( C ) and MCL-1 in each set ( D ). IL-27 prevented the decrease in osteoblast differentiation genes in mouse osteoblasts co-cultured with Ovx- CD4 + T cells, which include BMP-2 ( E ), Runx-2 ( F ), and Wnt-10b ( G ). IL-27 prevents secretion of osteoclastogenic cytokines from osteoblasts. Data are presented as mean ± S.E. ***, p
    Figure Legend Snippet: IL-27 inhibits mouse calvarial osteoblast apoptosis by enhancing Egr-2 expression and suppressing inhibitory effect of CD4 + T cells isolated from Ovx mice on osteoblast differentiation. A, representative images of apoptosis assay done by annexin and PI staining. B, percentage of apoptotic cells in each set. Transcript levels of Egr-2 ( C ) and MCL-1 in each set ( D ). IL-27 prevented the decrease in osteoblast differentiation genes in mouse osteoblasts co-cultured with Ovx- CD4 + T cells, which include BMP-2 ( E ), Runx-2 ( F ), and Wnt-10b ( G ). IL-27 prevents secretion of osteoclastogenic cytokines from osteoblasts. Data are presented as mean ± S.E. ***, p

    Techniques Used: Expressing, Isolation, Mouse Assay, Apoptosis Assay, Staining, Cell Culture

    IL-27 negatively regulates osteoclastogenesis through Egr-2-mediated regulation of Id2 protein. A, histomorphometric analysis of femoral epiphysis by TRAP staining. B, number of osteoclasts ( OC )/bone perimeter ( N.Oc/B.Pm ); C, osteoclast surface/bone surface (%) measured in all the groups. D, transcript levels of Egr-2 ; E, Id2 were determined in osteoclast cultures at days 2, 4, and 6. mRNA expression of EGR-2 ( F ), ID-2 ( G ), and TRAP expression ( H ) in EGR-2 knockdown osteoclasts by siRNA. Data are representative of three independent experiments. ***, p
    Figure Legend Snippet: IL-27 negatively regulates osteoclastogenesis through Egr-2-mediated regulation of Id2 protein. A, histomorphometric analysis of femoral epiphysis by TRAP staining. B, number of osteoclasts ( OC )/bone perimeter ( N.Oc/B.Pm ); C, osteoclast surface/bone surface (%) measured in all the groups. D, transcript levels of Egr-2 ; E, Id2 were determined in osteoclast cultures at days 2, 4, and 6. mRNA expression of EGR-2 ( F ), ID-2 ( G ), and TRAP expression ( H ) in EGR-2 knockdown osteoclasts by siRNA. Data are representative of three independent experiments. ***, p

    Techniques Used: Staining, Expressing

    Improved cortical bone parameters and bone biomechanical properties in Ovx mice treated with IL-27. A, representative 3D micro-CT images of cortical bone at the femoral diaphysis. B, Cs.Th was measured in all the groups. C, B.Ar was measured in all the groups. Three-point bending test of the femur was carried out to measure stiffness ( D ), energy ( E ) and power required to break femoral bone ( F ). n = 8 mice/group; data are presented as mean ± S.E.; ***, p
    Figure Legend Snippet: Improved cortical bone parameters and bone biomechanical properties in Ovx mice treated with IL-27. A, representative 3D micro-CT images of cortical bone at the femoral diaphysis. B, Cs.Th was measured in all the groups. C, B.Ar was measured in all the groups. Three-point bending test of the femur was carried out to measure stiffness ( D ), energy ( E ) and power required to break femoral bone ( F ). n = 8 mice/group; data are presented as mean ± S.E.; ***, p

    Techniques Used: Mouse Assay, Micro-CT

    5) Product Images from "IL-27 targets Foxp3+ Tregs to mediate antiinflammatory functions during experimental allergic airway inflammation"

    Article Title: IL-27 targets Foxp3+ Tregs to mediate antiinflammatory functions during experimental allergic airway inflammation

    Journal: JCI Insight

    doi: 10.1172/jci.insight.123216

    IL-27 inhibits effector T cell proliferation via Treg. ( A ) Foxp3 DTR mice were sensitized with CA in alum adjuvant and intranasally challenged in the presence or absence of IL-27 as described above. DTX was also injected as indicated. BrdU was injected 24 hours prior to sacrifice. BrdU incorporation was determined by FACS analysis as described in Methods. ( B ) Treg and effector T cell expression of the listed surface molecules was determined by FACS analysis. ( C ) Il27ra –/– mice were treated as described above. BrdU incorporation was determined by FACS analysis as described. The data shown represent the mean ± SD of 2 independent experiments ( n = 4–9). Each symbol represents an individually tested mouse. * P
    Figure Legend Snippet: IL-27 inhibits effector T cell proliferation via Treg. ( A ) Foxp3 DTR mice were sensitized with CA in alum adjuvant and intranasally challenged in the presence or absence of IL-27 as described above. DTX was also injected as indicated. BrdU was injected 24 hours prior to sacrifice. BrdU incorporation was determined by FACS analysis as described in Methods. ( B ) Treg and effector T cell expression of the listed surface molecules was determined by FACS analysis. ( C ) Il27ra –/– mice were treated as described above. BrdU incorporation was determined by FACS analysis as described. The data shown represent the mean ± SD of 2 independent experiments ( n = 4–9). Each symbol represents an individually tested mouse. * P

    Techniques Used: Mouse Assay, Injection, BrdU Incorporation Assay, FACS, Expressing

    Phosphorylation of STAT1 in response to IL-27 is reduced in T cells of asthmatic patients. ( A ) PBMCs collected from controls and asthmatic patients were ex vivo stimulated with or without IL-27 (10 ng/ml) for the indicated duration. STAT1 and STAT3 phosphorylation was determined by flow cytometry analysis. Treg and conventional CD4 T cells were gated as CD3 + CD4 + CD25 + CD127 lo/– and CD3 + CD4 + CD25 – CD127 lo/– cells, respectively. The geometric mean fluorescence intensity is normalized to those of untreated controls and shown as ΔgMFI. ( B ) IL-27Rα expression on Tregs and conventional CD4 T cells was determined by flow cytometry. All the experiments were repeated more than twice. The data shown indicate the mean ± SD ( n = 4–6). * P
    Figure Legend Snippet: Phosphorylation of STAT1 in response to IL-27 is reduced in T cells of asthmatic patients. ( A ) PBMCs collected from controls and asthmatic patients were ex vivo stimulated with or without IL-27 (10 ng/ml) for the indicated duration. STAT1 and STAT3 phosphorylation was determined by flow cytometry analysis. Treg and conventional CD4 T cells were gated as CD3 + CD4 + CD25 + CD127 lo/– and CD3 + CD4 + CD25 – CD127 lo/– cells, respectively. The geometric mean fluorescence intensity is normalized to those of untreated controls and shown as ΔgMFI. ( B ) IL-27Rα expression on Tregs and conventional CD4 T cells was determined by flow cytometry. All the experiments were repeated more than twice. The data shown indicate the mean ± SD ( n = 4–6). * P

    Techniques Used: Ex Vivo, Flow Cytometry, Cytometry, Fluorescence, Expressing

    Transfer of WT Tregs into Il27ra –/– mice diminishes allergic airway inflammation after IL-27 treatment. WT and Il27ra –/– mice were sensitized with CA in alum adjuvant and intranasally challenged in the presence or absence of intranasal IL-27. In some experiments, 2 × 10 6 iTregs were transferred 2 days before antigen challenge. Mice were sacrificed 24 hours after the last challenge. ( A ) BAL cells were examined for Ly6G and Siglec F expression, and differential cell counts was performed by FACS. ( B ) Lung cells were stimulated ex vivo, and intracellular cytokine expression was determined. ( C ) H E staining of the lung tissues are shown (original magnification ×20), and histology score was evaluated. The data shown represent the mean ± SD of 2 independent experiments ( n = 6). Each symbol represents an individually tested mouse. * P
    Figure Legend Snippet: Transfer of WT Tregs into Il27ra –/– mice diminishes allergic airway inflammation after IL-27 treatment. WT and Il27ra –/– mice were sensitized with CA in alum adjuvant and intranasally challenged in the presence or absence of intranasal IL-27. In some experiments, 2 × 10 6 iTregs were transferred 2 days before antigen challenge. Mice were sacrificed 24 hours after the last challenge. ( A ) BAL cells were examined for Ly6G and Siglec F expression, and differential cell counts was performed by FACS. ( B ) Lung cells were stimulated ex vivo, and intracellular cytokine expression was determined. ( C ) H E staining of the lung tissues are shown (original magnification ×20), and histology score was evaluated. The data shown represent the mean ± SD of 2 independent experiments ( n = 6). Each symbol represents an individually tested mouse. * P

    Techniques Used: Mouse Assay, Expressing, FACS, Ex Vivo, Staining

    Lag3 expression on Treg is necessary for Treg-mediated suppression in response to IL-27. ( A and B ) Foxp3 DTR mice were sensitized with CA in alum and intranasally challenged in the presence or absence of IL-27 as described above. DTX was injected 1 day before and on the day of first Ag challenge. Then, the mice received 2 × 10 6 in vitro–generated Foxp3 + Tregs deficient in Lag3 two days before antigen challenge. Mice were sacrificed 24 hours after the last challenge. Granulocytes and CD4 T cells in the BAL ( A ) and cytokine-expressing CD4 T cells in the lung ( B ) were calculated by FACS analysis. ( C ) Treg WT and Treg ΔIl27ra mice were sensitized with CA in alum and intranasally challenged as described above. Upon sacrifice, Lag3 and IL-10 expression of lung-infiltrating conventional CD4 and Foxp3 + Tregs was measured by FACS analysis. All the experiments were repeated more than twice. The data shown indicate the mean ± SD ( n = 4–13). Each symbol represents an individually tested animal. * P
    Figure Legend Snippet: Lag3 expression on Treg is necessary for Treg-mediated suppression in response to IL-27. ( A and B ) Foxp3 DTR mice were sensitized with CA in alum and intranasally challenged in the presence or absence of IL-27 as described above. DTX was injected 1 day before and on the day of first Ag challenge. Then, the mice received 2 × 10 6 in vitro–generated Foxp3 + Tregs deficient in Lag3 two days before antigen challenge. Mice were sacrificed 24 hours after the last challenge. Granulocytes and CD4 T cells in the BAL ( A ) and cytokine-expressing CD4 T cells in the lung ( B ) were calculated by FACS analysis. ( C ) Treg WT and Treg ΔIl27ra mice were sensitized with CA in alum and intranasally challenged as described above. Upon sacrifice, Lag3 and IL-10 expression of lung-infiltrating conventional CD4 and Foxp3 + Tregs was measured by FACS analysis. All the experiments were repeated more than twice. The data shown indicate the mean ± SD ( n = 4–13). Each symbol represents an individually tested animal. * P

    Techniques Used: Expressing, Mouse Assay, Injection, In Vitro, Generated, FACS

    Intranasal IL-27 administration attenuates the development of allergic airway inflammation. ( A ) Experimental protocol. B6 mice were injected i.p. on days 0 and 7 with CA in alum adjuvant. Starting at day 14, mice were intranasally challenged for 4 consecutive days with CA alone or together with IL-27. Mice were sacrificed 24 hours after the last challenge. ( B ) BAL cells were examined for Ly6G and Siglec F expression. Differential cell count was performed by FACS analysis. ( C and D ) Lung ( C ) and draining medLN ( D ) cells were harvested and stimulated ex vivo to assess intracellular cytokine expression. ( E ) IL-4 and IL-13 secretion in the BALF was determined using a CBA assay. ( F ) H E and PAS staining of the lung tissues (original magnification ×20 and ×10, respectively) was used to evaluate inflammation. Histology score was determined as described in Methods. ( G ) Muc5a and Muc5b mRNA expression in the lung was determined by qPCR analysis. ( H ) Airway resistance was measured by flexivent experiments. Each symbol represents individually tested animal. The data shown are the mean ± SD of 3 independent experiments ( n = 9). * P
    Figure Legend Snippet: Intranasal IL-27 administration attenuates the development of allergic airway inflammation. ( A ) Experimental protocol. B6 mice were injected i.p. on days 0 and 7 with CA in alum adjuvant. Starting at day 14, mice were intranasally challenged for 4 consecutive days with CA alone or together with IL-27. Mice were sacrificed 24 hours after the last challenge. ( B ) BAL cells were examined for Ly6G and Siglec F expression. Differential cell count was performed by FACS analysis. ( C and D ) Lung ( C ) and draining medLN ( D ) cells were harvested and stimulated ex vivo to assess intracellular cytokine expression. ( E ) IL-4 and IL-13 secretion in the BALF was determined using a CBA assay. ( F ) H E and PAS staining of the lung tissues (original magnification ×20 and ×10, respectively) was used to evaluate inflammation. Histology score was determined as described in Methods. ( G ) Muc5a and Muc5b mRNA expression in the lung was determined by qPCR analysis. ( H ) Airway resistance was measured by flexivent experiments. Each symbol represents individually tested animal. The data shown are the mean ± SD of 3 independent experiments ( n = 9). * P

    Techniques Used: Mouse Assay, Injection, Expressing, Cell Counting, FACS, Ex Vivo, Crocin Bleaching Assay, Staining, Real-time Polymerase Chain Reaction

    6) Product Images from "IL-6-dependent and -independent pathways in the development of interleukin 17-producing T helper cells"

    Article Title: IL-6-dependent and -independent pathways in the development of interleukin 17-producing T helper cells

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

    doi: 10.1073/pnas.0705268104

    Activation of Stat1 and Stat3 by stimulation to produce IL-17 in naïve T cells. MACS-sorted naïve T cells were cultured with anti-CD3/CD28 beads. ( A ) Naïve T cells were stimulated with TGF-β alone or TGF-β plus IL-6 for 30 min or 24 h. ( B ) Naïve T cells were stimulated with TGF-β alone or TGF-β plus IL-6 in the presence or absence of IL-27 or IFN-γ for 24 h. ( C ) Naïve T cells were cultured with DCCM with or without IL-27. Cells were fixed and permeabilized in 90% methanol, followed by staining with Alexa Fluor 488-conjugated phospho-Stat1 and phycoerythrin-conjugated phospho-Stat3. Intracellular phospho-Stat1 and -Stat3 were measured by flow cytometry. These results are representative of three independent experiments.
    Figure Legend Snippet: Activation of Stat1 and Stat3 by stimulation to produce IL-17 in naïve T cells. MACS-sorted naïve T cells were cultured with anti-CD3/CD28 beads. ( A ) Naïve T cells were stimulated with TGF-β alone or TGF-β plus IL-6 for 30 min or 24 h. ( B ) Naïve T cells were stimulated with TGF-β alone or TGF-β plus IL-6 in the presence or absence of IL-27 or IFN-γ for 24 h. ( C ) Naïve T cells were cultured with DCCM with or without IL-27. Cells were fixed and permeabilized in 90% methanol, followed by staining with Alexa Fluor 488-conjugated phospho-Stat1 and phycoerythrin-conjugated phospho-Stat3. Intracellular phospho-Stat1 and -Stat3 were measured by flow cytometry. These results are representative of three independent experiments.

    Techniques Used: Activation Assay, Magnetic Cell Separation, Cell Culture, Staining, Flow Cytometry, Cytometry

    IL-17 is produced by DCCM without IL-6-mediated activity in naïve T cells. ( A ) Purified naïve T cells were cultured with anti-CD3/CD28 beads and the indicated cytokines, LPS, or CM from DCs stimulated with LPS. ( B ) Naïve T cells were stimulated with TGF-β plus IL-6, DCCM, or DCCM from IL-6 KO mice in the presence or absence of IL-27 or neutralizing antibodies for IL-6. Supernatants were collected 2 days after stimulation, and IL-17 production was measured by means of ELISA. Data show means ± SE of three independent experiments. (C) MACS-sorted naïve T cells were activated with anti-CD3/CD28 beads. Cells were cultured with the indicated stimulators. After 3 days, cells were restimulated with PMA and ionomycin before intracellular cytokine staining for IFN-γ and IL-17 and analysis by flow cytometry. Dot plots show intracellular staining for IFN-γ and IL-17.
    Figure Legend Snippet: IL-17 is produced by DCCM without IL-6-mediated activity in naïve T cells. ( A ) Purified naïve T cells were cultured with anti-CD3/CD28 beads and the indicated cytokines, LPS, or CM from DCs stimulated with LPS. ( B ) Naïve T cells were stimulated with TGF-β plus IL-6, DCCM, or DCCM from IL-6 KO mice in the presence or absence of IL-27 or neutralizing antibodies for IL-6. Supernatants were collected 2 days after stimulation, and IL-17 production was measured by means of ELISA. Data show means ± SE of three independent experiments. (C) MACS-sorted naïve T cells were activated with anti-CD3/CD28 beads. Cells were cultured with the indicated stimulators. After 3 days, cells were restimulated with PMA and ionomycin before intracellular cytokine staining for IFN-γ and IL-17 and analysis by flow cytometry. Dot plots show intracellular staining for IFN-γ and IL-17.

    Techniques Used: Produced, Activity Assay, Purification, Cell Culture, Mouse Assay, Enzyme-linked Immunosorbent Assay, Magnetic Cell Separation, Staining, Flow Cytometry, Cytometry

    RORγt mRNA expression by IL-6 and TGF-β with or without IL-27 or IFN-γ. Isolated naïve T cells were stimulated with anti-CD3/CD28 beads and the indicated cytokines, followed by total RNA and cDNA prepared as described in Materials and Methods . RORγt induction was examined by using RT-PCR.
    Figure Legend Snippet: RORγt mRNA expression by IL-6 and TGF-β with or without IL-27 or IFN-γ. Isolated naïve T cells were stimulated with anti-CD3/CD28 beads and the indicated cytokines, followed by total RNA and cDNA prepared as described in Materials and Methods . RORγt induction was examined by using RT-PCR.

    Techniques Used: Expressing, Isolation, Reverse Transcription Polymerase Chain Reaction

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  • 93
    R&D Systems recombinant mouse il 27
    Deletion of p28-coding gene in DCs. ( A ) Mouse genotyping was performed using PCR. ( B ) p28 and Ebi3 mRNA levels were determined by quantitative PCR in total thymic DCs form CD11c-cre p28 flox/flox and WT mice. The <t>IL-27</t> protein expression was measured by flow cytometry. ( C ) IL-27RA and gp130 mRNA expression was compared in CD4 SP thymocytes form CD11c-cre p28 flox/flox and WT mice. The experiments shown in B and C were repeated for more than three times and the data are presented as mean ± s.d. ** p
    Recombinant Mouse Il 27, 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/recombinant mouse il 27/product/R&D Systems
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant mouse il 27 - by Bioz Stars, 2021-05
    93/100 stars
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    96
    R&D Systems il 27
    TLR3 co‐stimulation of microglia does not lead to overshooting responses. (a–k) WT microglial cells were stimulated for 24 hr with TLR2 or TLR3 agonists alone or in combination. Cell culture supernatants were collected and cytokine production was measured by multiplex assay (IL‐12p70, IL‐23, <t>IL‐27,</t> IL‐22, IFN‐γ, IL‐4, and IL‐5) or ELISA (TNF, IL‐6, IFN‐β, and IL‐1β). Significant statistical differences relative to TLR2 are represented by *; to TLR3 by #. (l) WT and IFNAR−/− microglial cells were stimulated for 24 hr with the TLR2 or TLR3 agonists alone or in combination. Cell culture supernatants were collected and IL‐27 production was measured by multiplex. (m) WT microglial cells were stimulated for 24 hr with the TLR2 agonist, IFN‐β or IL‐27 alone or in combination. Cell culture supernatants were collected and IL‐10 production was measured by ELISA. Significant statistical differences relative to TLR2 are represented by *; to TLR2 + IFN‐β by #; to TLR2 + IL‐27 by $. The detection limit for each cytokine is represented as a dotted line in each graph. Represented are the mean ± SD for triplicate wells per condition set after mixed cultures generated from independent mice. Statistical differences were assessed by student's t test or one‐way ANOVA. Significant statistical differences are represented by one symbol, p
    Il 27, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il 27/product/R&D Systems
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 27 - by Bioz Stars, 2021-05
    96/100 stars
      Buy from Supplier

    92
    R&D Systems il 17d
    Expression of <t>IL-17D</t> mediates progressor tumor rejection
    Il 17d, 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/il 17d/product/R&D Systems
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 17d - by Bioz Stars, 2021-05
    92/100 stars
      Buy from Supplier

    N/A
    The Recombinant Mouse IL 27 p28 IL 30 Protein from R D Systems is derived from NS0 The Recombinant Mouse IL 27 p28 IL 30 Protein has been validated for
      Buy from Supplier

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    Deletion of p28-coding gene in DCs. ( A ) Mouse genotyping was performed using PCR. ( B ) p28 and Ebi3 mRNA levels were determined by quantitative PCR in total thymic DCs form CD11c-cre p28 flox/flox and WT mice. The IL-27 protein expression was measured by flow cytometry. ( C ) IL-27RA and gp130 mRNA expression was compared in CD4 SP thymocytes form CD11c-cre p28 flox/flox and WT mice. The experiments shown in B and C were repeated for more than three times and the data are presented as mean ± s.d. ** p

    Journal: Scientific Reports

    Article Title: Thymic DCs derived IL-27 regulates the final maturation of CD4+ SP thymocytes

    doi: 10.1038/srep30448

    Figure Lengend Snippet: Deletion of p28-coding gene in DCs. ( A ) Mouse genotyping was performed using PCR. ( B ) p28 and Ebi3 mRNA levels were determined by quantitative PCR in total thymic DCs form CD11c-cre p28 flox/flox and WT mice. The IL-27 protein expression was measured by flow cytometry. ( C ) IL-27RA and gp130 mRNA expression was compared in CD4 SP thymocytes form CD11c-cre p28 flox/flox and WT mice. The experiments shown in B and C were repeated for more than three times and the data are presented as mean ± s.d. ** p

    Article Snippet: Recombinant mouse IL-27 and IL-27 neutralizing antibody (Anti-IL-27p28) were purchased from R & D Systems (Minneapolis, MN).

    Techniques: Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Mouse Assay, Expressing, Flow Cytometry, Cytometry

    The rescuing effect of intrathymically injected IL-27. CD11c-cre p28 flox/flox and WT mice received intrathymic injection of IL-27 or PBS. Two days later, the thymocytes were collected, stained with antibodies for CD4, CD8, CD69, Qa-2 and analyzed by flow cytometry. ( A ) Representative dot plots showing CD69 and Qa-2 expression in CD4 SP thymocytes. ( B ) The percentage of CD4 SP cells in total thymocytes, the percentage of SP4 cells in the CD4 SP population, and the absolute number of SP4 cells are shown. Each symbol represents an individual sample and the horizontal bar denotes the mean. * p

    Journal: Scientific Reports

    Article Title: Thymic DCs derived IL-27 regulates the final maturation of CD4+ SP thymocytes

    doi: 10.1038/srep30448

    Figure Lengend Snippet: The rescuing effect of intrathymically injected IL-27. CD11c-cre p28 flox/flox and WT mice received intrathymic injection of IL-27 or PBS. Two days later, the thymocytes were collected, stained with antibodies for CD4, CD8, CD69, Qa-2 and analyzed by flow cytometry. ( A ) Representative dot plots showing CD69 and Qa-2 expression in CD4 SP thymocytes. ( B ) The percentage of CD4 SP cells in total thymocytes, the percentage of SP4 cells in the CD4 SP population, and the absolute number of SP4 cells are shown. Each symbol represents an individual sample and the horizontal bar denotes the mean. * p

    Article Snippet: Recombinant mouse IL-27 and IL-27 neutralizing antibody (Anti-IL-27p28) were purchased from R & D Systems (Minneapolis, MN).

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

    IL-27-enhanced development of Qa-2 + cells in CD4 + SP1 thymocyte cultures. ( A ) The profile of isolated CD4 + SP1 thymocytes was shown by the expression of 6C10 and CD69 (left), or by the expression of CD69 and Qa-2 (right). The purity of the sorted cells was 97–99%. ( B ) Freshly isolated CD4 + SP1 thymocytes from WT mice were cultured for 4 days in RPMI-1640 medium with or without the addition of IL-27, IL-30 or anti-IL-27p28. Acquisition of Qa-2 expression was detected by surface staining. Representative dot plots (left) and the percentage of Qa-2 + cells as well as the recovered live cells number (right) were presented. ( C ) SP1 cells were co-cultured with thymic DCs obtained from CD11c-cre p28 flox/flox or WT mice for 4 days in the presence or absence of IL-27. Qa-2 expression was evaluated by FACS analysis. Representative dot plots (left) and the percentage of Qa-2 + cells (right) are presented. The experiments were repeated for five times. * p

    Journal: Scientific Reports

    Article Title: Thymic DCs derived IL-27 regulates the final maturation of CD4+ SP thymocytes

    doi: 10.1038/srep30448

    Figure Lengend Snippet: IL-27-enhanced development of Qa-2 + cells in CD4 + SP1 thymocyte cultures. ( A ) The profile of isolated CD4 + SP1 thymocytes was shown by the expression of 6C10 and CD69 (left), or by the expression of CD69 and Qa-2 (right). The purity of the sorted cells was 97–99%. ( B ) Freshly isolated CD4 + SP1 thymocytes from WT mice were cultured for 4 days in RPMI-1640 medium with or without the addition of IL-27, IL-30 or anti-IL-27p28. Acquisition of Qa-2 expression was detected by surface staining. Representative dot plots (left) and the percentage of Qa-2 + cells as well as the recovered live cells number (right) were presented. ( C ) SP1 cells were co-cultured with thymic DCs obtained from CD11c-cre p28 flox/flox or WT mice for 4 days in the presence or absence of IL-27. Qa-2 expression was evaluated by FACS analysis. Representative dot plots (left) and the percentage of Qa-2 + cells (right) are presented. The experiments were repeated for five times. * p

    Article Snippet: Recombinant mouse IL-27 and IL-27 neutralizing antibody (Anti-IL-27p28) were purchased from R & D Systems (Minneapolis, MN).

    Techniques: Isolation, Expressing, Mouse Assay, Cell Culture, Staining, FACS

    IL27- mediated signal in CD4 + SP thymocytes. ( A ) CD4 + SP thymocytes were isolated form CD11c-cre p28 flox/flox and WT mice, cultured in RPMI-1640 medium with IL-27. Cells were harvested at 0, 5, 15, 30 and 60 min after stimulation, Phosphorylated and total STAT1 and STAT3 were detected by Western blotting. The cropped gels have been run under the same experimental conditions. Representative blots are shown of three independent experiments. β-actin was used as a control. ( B ) SP3 cells from CD11c-cre p28 flox/flox mice and WT littermates were stimulated by 2 ng/ml IL-27 for 24 hours, cells were collected and the mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 were evaluated (n ≥ 3). The experiments were repeated for three times and the data are shown as mean ± s.d. ( C ) Total RNA was extracted from FACS-purified CD4 + SP thymocyte subsets from CD11c-cre p28 flox/flox mice and WT littermates. Quantitative PCR was performed to analyze mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 (n ≥ 3). * p

    Journal: Scientific Reports

    Article Title: Thymic DCs derived IL-27 regulates the final maturation of CD4+ SP thymocytes

    doi: 10.1038/srep30448

    Figure Lengend Snippet: IL27- mediated signal in CD4 + SP thymocytes. ( A ) CD4 + SP thymocytes were isolated form CD11c-cre p28 flox/flox and WT mice, cultured in RPMI-1640 medium with IL-27. Cells were harvested at 0, 5, 15, 30 and 60 min after stimulation, Phosphorylated and total STAT1 and STAT3 were detected by Western blotting. The cropped gels have been run under the same experimental conditions. Representative blots are shown of three independent experiments. β-actin was used as a control. ( B ) SP3 cells from CD11c-cre p28 flox/flox mice and WT littermates were stimulated by 2 ng/ml IL-27 for 24 hours, cells were collected and the mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 were evaluated (n ≥ 3). The experiments were repeated for three times and the data are shown as mean ± s.d. ( C ) Total RNA was extracted from FACS-purified CD4 + SP thymocyte subsets from CD11c-cre p28 flox/flox mice and WT littermates. Quantitative PCR was performed to analyze mRNA expression of IFIT1, IFIT3, IRF7 and IRF8 (n ≥ 3). * p

    Article Snippet: Recombinant mouse IL-27 and IL-27 neutralizing antibody (Anti-IL-27p28) were purchased from R & D Systems (Minneapolis, MN).

    Techniques: Isolation, Mouse Assay, Cell Culture, Western Blot, Expressing, FACS, Purification, Real-time Polymerase Chain Reaction

    IL-27 and its receptor expression in the thymus. ( A ) Total thymocyte, CD11c int CD45RA + pDC, CD11c hi CD8α + CD172α − and CD11c hi CD8α − CD172α + cDC, CD45 − Epcam hi Ly51 + cTEC, CD45 - Epcam hi Ly51 - mTEC and CD11b + F4/80 + macrophage were isolated from C57BL/6J mice. The expression of p28 and EBI3 mRNA was evaluated by quantitative PCR. nd, no detectable. ( B ) IL-27 protein expression in pDC, CD8α + and CD172α + cDC was analyzed by flow cytometry following intracellular staining with anti-IL-27. Representative histograms are shown on the left and the mean fluorescence intensity (MFI) on the right. ( C ) Purified DN, DP, CD8 SP and the four subsets of CD4 SP (SP1-4) thymocytes were analyzed for IL-27RA and gp130 mRNA expression using quantitative PCR. The lymph node (LN) naïve T cell was used as a positive control. All the experiments were repeated for more than three times and the data are shown as mean ± s.d. * p

    Journal: Scientific Reports

    Article Title: Thymic DCs derived IL-27 regulates the final maturation of CD4+ SP thymocytes

    doi: 10.1038/srep30448

    Figure Lengend Snippet: IL-27 and its receptor expression in the thymus. ( A ) Total thymocyte, CD11c int CD45RA + pDC, CD11c hi CD8α + CD172α − and CD11c hi CD8α − CD172α + cDC, CD45 − Epcam hi Ly51 + cTEC, CD45 - Epcam hi Ly51 - mTEC and CD11b + F4/80 + macrophage were isolated from C57BL/6J mice. The expression of p28 and EBI3 mRNA was evaluated by quantitative PCR. nd, no detectable. ( B ) IL-27 protein expression in pDC, CD8α + and CD172α + cDC was analyzed by flow cytometry following intracellular staining with anti-IL-27. Representative histograms are shown on the left and the mean fluorescence intensity (MFI) on the right. ( C ) Purified DN, DP, CD8 SP and the four subsets of CD4 SP (SP1-4) thymocytes were analyzed for IL-27RA and gp130 mRNA expression using quantitative PCR. The lymph node (LN) naïve T cell was used as a positive control. All the experiments were repeated for more than three times and the data are shown as mean ± s.d. * p

    Article Snippet: Recombinant mouse IL-27 and IL-27 neutralizing antibody (Anti-IL-27p28) were purchased from R & D Systems (Minneapolis, MN).

    Techniques: Expressing, Isolation, Mouse Assay, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry, Staining, Fluorescence, Purification, Positive Control

    TLR3 co‐stimulation of microglia does not lead to overshooting responses. (a–k) WT microglial cells were stimulated for 24 hr with TLR2 or TLR3 agonists alone or in combination. Cell culture supernatants were collected and cytokine production was measured by multiplex assay (IL‐12p70, IL‐23, IL‐27, IL‐22, IFN‐γ, IL‐4, and IL‐5) or ELISA (TNF, IL‐6, IFN‐β, and IL‐1β). Significant statistical differences relative to TLR2 are represented by *; to TLR3 by #. (l) WT and IFNAR−/− microglial cells were stimulated for 24 hr with the TLR2 or TLR3 agonists alone or in combination. Cell culture supernatants were collected and IL‐27 production was measured by multiplex. (m) WT microglial cells were stimulated for 24 hr with the TLR2 agonist, IFN‐β or IL‐27 alone or in combination. Cell culture supernatants were collected and IL‐10 production was measured by ELISA. Significant statistical differences relative to TLR2 are represented by *; to TLR2 + IFN‐β by #; to TLR2 + IL‐27 by $. The detection limit for each cytokine is represented as a dotted line in each graph. Represented are the mean ± SD for triplicate wells per condition set after mixed cultures generated from independent mice. Statistical differences were assessed by student's t test or one‐way ANOVA. Significant statistical differences are represented by one symbol, p

    Journal: Glia

    Article Title: Interferon‐β regulates the production of IL‐10 by toll‐like receptor‐activated microglia. Interferon‐ β regulates the production of IL‐10 by toll‐like receptor‐activated microglia

    doi: 10.1002/glia.23172

    Figure Lengend Snippet: TLR3 co‐stimulation of microglia does not lead to overshooting responses. (a–k) WT microglial cells were stimulated for 24 hr with TLR2 or TLR3 agonists alone or in combination. Cell culture supernatants were collected and cytokine production was measured by multiplex assay (IL‐12p70, IL‐23, IL‐27, IL‐22, IFN‐γ, IL‐4, and IL‐5) or ELISA (TNF, IL‐6, IFN‐β, and IL‐1β). Significant statistical differences relative to TLR2 are represented by *; to TLR3 by #. (l) WT and IFNAR−/− microglial cells were stimulated for 24 hr with the TLR2 or TLR3 agonists alone or in combination. Cell culture supernatants were collected and IL‐27 production was measured by multiplex. (m) WT microglial cells were stimulated for 24 hr with the TLR2 agonist, IFN‐β or IL‐27 alone or in combination. Cell culture supernatants were collected and IL‐10 production was measured by ELISA. Significant statistical differences relative to TLR2 are represented by *; to TLR2 + IFN‐β by #; to TLR2 + IL‐27 by $. The detection limit for each cytokine is represented as a dotted line in each graph. Represented are the mean ± SD for triplicate wells per condition set after mixed cultures generated from independent mice. Statistical differences were assessed by student's t test or one‐way ANOVA. Significant statistical differences are represented by one symbol, p

    Article Snippet: Mouse recombinant IFN‐β, IL‐27, IFN‐γ, and IL‐17 were bought from R & D systems.

    Techniques: Cell Culture, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Generated, Mouse Assay

    Inflammatory landscape of TLR‐stimulated microglia. (a–k). Primary microglial cell cultures were left unstimulated or stimulated for 24 hr with chemical TLR agonists for TLR2, TLR3, TLR4, or TLR9, as described in Section 2 . Cell culture supernatants were collected and cytokine production was measured by multiplex assay (IL‐10, TNF, IL‐12p70, IL‐23, IL‐27, IL‐22, IFN‐γ, IL‐4, and IL‐5) or ELISA (IL‐6 and IFN‐β). Unstimulated cells did not produce detectable amounts of cytokines. The detection limit for each cytokine is represented as a dotted line in each graph. Represented are the mean ± SD for triplicate wells per condition set after mixed cultures generated from independent mice. Statistical differences were assessed by one‐way ANOVA or student's t test. Significant statistical differences relative to TLR2 are represented by *; to TLR3 by #; to TLR4 by $. One symbol, p

    Journal: Glia

    Article Title: Interferon‐β regulates the production of IL‐10 by toll‐like receptor‐activated microglia. Interferon‐ β regulates the production of IL‐10 by toll‐like receptor‐activated microglia

    doi: 10.1002/glia.23172

    Figure Lengend Snippet: Inflammatory landscape of TLR‐stimulated microglia. (a–k). Primary microglial cell cultures were left unstimulated or stimulated for 24 hr with chemical TLR agonists for TLR2, TLR3, TLR4, or TLR9, as described in Section 2 . Cell culture supernatants were collected and cytokine production was measured by multiplex assay (IL‐10, TNF, IL‐12p70, IL‐23, IL‐27, IL‐22, IFN‐γ, IL‐4, and IL‐5) or ELISA (IL‐6 and IFN‐β). Unstimulated cells did not produce detectable amounts of cytokines. The detection limit for each cytokine is represented as a dotted line in each graph. Represented are the mean ± SD for triplicate wells per condition set after mixed cultures generated from independent mice. Statistical differences were assessed by one‐way ANOVA or student's t test. Significant statistical differences relative to TLR2 are represented by *; to TLR3 by #; to TLR4 by $. One symbol, p

    Article Snippet: Mouse recombinant IFN‐β, IL‐27, IFN‐γ, and IL‐17 were bought from R & D systems.

    Techniques: Cell Culture, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Generated, Mouse Assay

    Expression of IL-17D mediates progressor tumor rejection

    Journal: Cell reports

    Article Title: Interleukin-17D mediates tumor rejection through recruitment of natural killer cells

    doi: 10.1016/j.celrep.2014.03.073

    Figure Lengend Snippet: Expression of IL-17D mediates progressor tumor rejection

    Article Snippet: On day 7, either 1ml of LPS (1μg/ml), IL-17A (5μg/ml) (R & D Systems), IL-17D (5μg/ml) (R & D Systems), IL-17D (5μg/ml) (Mayfield Lab), MCP-1 (5μg/ml) (Peprotech), or IL-17D (5μg/ml) + anti-MCP-1 polyclonal antibodies (25μg/ml) (R & D Systems) was injected into mouse air pouches 8h before air pouch harvest.

    Techniques: Expressing

    IL-17D is highly expressed in some regressor cell lines and is downregulated in progressor tumor cell lines and several human cancer samples

    Journal: Cell reports

    Article Title: Interleukin-17D mediates tumor rejection through recruitment of natural killer cells

    doi: 10.1016/j.celrep.2014.03.073

    Figure Lengend Snippet: IL-17D is highly expressed in some regressor cell lines and is downregulated in progressor tumor cell lines and several human cancer samples

    Article Snippet: On day 7, either 1ml of LPS (1μg/ml), IL-17A (5μg/ml) (R & D Systems), IL-17D (5μg/ml) (R & D Systems), IL-17D (5μg/ml) (Mayfield Lab), MCP-1 (5μg/ml) (Peprotech), or IL-17D (5μg/ml) + anti-MCP-1 polyclonal antibodies (25μg/ml) (R & D Systems) was injected into mouse air pouches 8h before air pouch harvest.

    Techniques:

    Overexpression of IL-17D in progressor tumors recruits NK cells that are required for tumor rejection in WT mice and promote M1 macrophage infiltration

    Journal: Cell reports

    Article Title: Interleukin-17D mediates tumor rejection through recruitment of natural killer cells

    doi: 10.1016/j.celrep.2014.03.073

    Figure Lengend Snippet: Overexpression of IL-17D in progressor tumors recruits NK cells that are required for tumor rejection in WT mice and promote M1 macrophage infiltration

    Article Snippet: On day 7, either 1ml of LPS (1μg/ml), IL-17A (5μg/ml) (R & D Systems), IL-17D (5μg/ml) (R & D Systems), IL-17D (5μg/ml) (Mayfield Lab), MCP-1 (5μg/ml) (Peprotech), or IL-17D (5μg/ml) + anti-MCP-1 polyclonal antibodies (25μg/ml) (R & D Systems) was injected into mouse air pouches 8h before air pouch harvest.

    Techniques: Over Expression, Mouse Assay

    Recombinant mouse IL-17D recruits NK cells in an air pouch inflammation model

    Journal: Cell reports

    Article Title: Interleukin-17D mediates tumor rejection through recruitment of natural killer cells

    doi: 10.1016/j.celrep.2014.03.073

    Figure Lengend Snippet: Recombinant mouse IL-17D recruits NK cells in an air pouch inflammation model

    Article Snippet: On day 7, either 1ml of LPS (1μg/ml), IL-17A (5μg/ml) (R & D Systems), IL-17D (5μg/ml) (R & D Systems), IL-17D (5μg/ml) (Mayfield Lab), MCP-1 (5μg/ml) (Peprotech), or IL-17D (5μg/ml) + anti-MCP-1 polyclonal antibodies (25μg/ml) (R & D Systems) was injected into mouse air pouches 8h before air pouch harvest.

    Techniques: Recombinant