il-10 Search Results


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  • 99
    Thermo Fisher il 10
    <t>IL-10</t> expression supports EMT in invasive LSCC. (a) IL-10 plasmatic concentrations by ELISA in healthy donors (HD) ( n = 9) and LSCC patients ( n = 20), and Mann–Whitney U test performed for statistical analysis; data are shown as mean ± SD, ∗ p ≤ 0.05. (b) Representative IL-10 and CD68 stainings in laryngeal tissues (magnification ×40) (yellow arrow: normal epithelium, black arrow: tumor cells, and green arrow: stomal cells). (c) Percentage of stromal cells producing IL-10 was evaluated by IHC and correlated to the percentage of CD68 + stromal macrophage by the Pearson correlation test. (d) Correlogram: EMT biomarkers expression levels versus IL-10 and NOS2 expression in invasive LSCC ( n = 20).
    Il 10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2937 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems il 10
    4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) recovered high-fat diet (HFD)-impaired insulin signaling and inflammation in visceral adipose (VAD) tissue of LDLR −/− mice. ( A ) The peripheral-tissue insulin signaling pathway [insulin receptor, pyruvate dehydrogenase kinase isozyme 1 (PDK1), Akt, glycogen synthase kinase 3β (GSK3β)] was analyzed by western immunoblots. ( B ) Quantification of insulin receptor-α and -β expression, measured by imageQuant. Amounts of phosphorylated and total PDK1 ( C ), Akt ( D ), and GSK3β ( E ). ( F ) VAD tissue inflammatory cytokines and macrophage markers [interleukin (IL)-6, <t>IL-10,</t> CD11b, F4/80] were analyzed by western immunoblots, with protein levels quantified by imageQuant. ( G ) Tumor necrosis factor-α (TNF-α) expression in VAD tissue was analyzed by ELISA. ( H ) Serine protease activity in VAD tissue of mice receiving the chow, HFD, and HFD + AEBSF treatments ( n = 6/group). * P
    Il 10, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 3515 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gene exp il10 mm00439614 m1
    Construction and validation of Lenti-IL-27HA and Lenti-IL-35HA. ( A , B ). Graphic representation of the lentiviral constructs used to express IL-27 ( A ) and IL-35 ( B ). ( C , D ) Cytokines production was assessed by ELISA (IL-27, C ) or WB (IL-35, D ). ( E – G ). Affinity purified IL-27 (white bars) and IL-35 (black bars) were used to treat polarized CD4 + T cells and measure mRNA expression of IL-2 ( E ), IL-17 ( F ), gm-csf ( G ), and <t>IL-10</t> ( F ) (mean ± sd). NC = no cytokines. *p
    Gene Exp Il10 Mm00439614 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 830 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson anti il 10
    <t>IL-10–producing</t> Tfh cells emerge after vaccination, and IL-10, but neither T regs nor Tfr cells, limits Tfh-dependent vaccine responses in aged mice. ( A ) Plots show IA b -NP tetramer staining in immunized and unimmunized young (3.5 months, n = 5) and aged (18 months, n = 5) IL-10GFP–FOXP3RFP dual-reporter mice. Graphs show the frequency and number of NP-specific FoxP3 − CD4 + T cells that are IL-10 + and the number of NP-specific FoxP3 − IL-10 + T cells that are CXCR5 + PD1 + (means ± SEM). ( B ) Aged (17 months, n ≥ 7) WT C57BL/6 (isotype or anti–IL-10) and (20 months, n = 5) FoxP3 − DTR [phosphate-buffered saline (PBS)– or DT-treated] mice were immunized with nitrophenol-keyhole limpet hemocyanin (NP-KLH) in alum. Plots display the frequency of germinal center (GC) B cells (NP-specific) gated on Fas + GL7 + . Graphs show the frequency and number of B cells that are IgG1 + NP + (means ± SEM) and serum levels of immunoglobulin (Ig) specific for NP (IgG1) (means ± SEM). Data are pooled from two independent experiments. ( C to E ) Tfh10 cells accumulate during aging in humans. Human spleen cells from young (Y) (median, 18.8; range, 18 to 26 years; three males and five females) and old (O) (median, 62; range, 60 to 67 years; four males and four females) individuals were analyzed by flow cytometry. (C) The frequencies of CD3 + CD4 + CD45RO + FoxP3 − that are CXCR5 + PD1 + (means ± SEM). (D and E) CD4 + T cells were bead-purified by negative selection; FACS-sorted memory CD4 + T cells (CD45RO + ) into Tfh cells (CD25 − CD127 + PD1 + CXCR5 + ), T regs (CD25 + CD127 − PD1 − CXCR5 − ), and other memory cells (CD25 − CD127 + PD1 − CXCR5 − ) were either stimulated in vitro with anti-CD3/CD28 beads or unstimulated, and cytokines were analyzed by Luminex (means ± SEM). Each individual is represented by a symbol. * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001, Student’s t test. BSS, balanced salt solution.
    Anti Il 10, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 964 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gene exp il10 mm01288386 m1
    Macrophages from myeloid-deficient MOF mice demonstrate decreased NF-κB inflammatory genes. ( A ) Representative figures showing Mof deletion in BMDMs and wound macrophages from Lyz2Cre Mof fl/fl mice. BMDMs were derived from Lyz2Cre Mof fl/fl or Mof fl/fl mice. Wound macrophages were sorted by magnetic-activated cell sorting (MACS) from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Mof gene expression was measured by qPCR ( n = 3 × 4 mice pooled/replicate, repeated twice). ( B ) Representative figures showing inflammatory cytokine gene expression in LPS-stimulated BMDMs. BMDMs were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice and treated with LPS or vehicle control for 8 hours. Il1b , Tnf , and Nos2 gene expression was measured by qPCR ( n = 3, repeated twice). ( C ) Representative figures showing baseline inflammatory cytokine gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Il1b ( n = 3), Tnf ( n = 3), and Nos2 ( n = 4) expression was measured by qPCR ( n = 3 mice pooled/replicate, repeated twice). ( D ) Representative figures showing antiinflammatory gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. <t>Il10</t> , Arg1 , and Ym1 expression was measured by qPCR ( n = 3 × 3 mice pooled/replicate, repeated twice). A , C , and D were analyzed using a 2-tailed Student’s t test. B was analyzed using 2-way ANOVA followed by Holm-Šídák test for multiple comparisons.
    Gene Exp Il10 Mm01288386 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 414 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gene exp il10 mm00439616 m1
    Macrophages from myeloid-deficient MOF mice demonstrate decreased NF-κB inflammatory genes. ( A ) Representative figures showing Mof deletion in BMDMs and wound macrophages from Lyz2Cre Mof fl/fl mice. BMDMs were derived from Lyz2Cre Mof fl/fl or Mof fl/fl mice. Wound macrophages were sorted by magnetic-activated cell sorting (MACS) from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Mof gene expression was measured by qPCR ( n = 3 × 4 mice pooled/replicate, repeated twice). ( B ) Representative figures showing inflammatory cytokine gene expression in LPS-stimulated BMDMs. BMDMs were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice and treated with LPS or vehicle control for 8 hours. Il1b , Tnf , and Nos2 gene expression was measured by qPCR ( n = 3, repeated twice). ( C ) Representative figures showing baseline inflammatory cytokine gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Il1b ( n = 3), Tnf ( n = 3), and Nos2 ( n = 4) expression was measured by qPCR ( n = 3 mice pooled/replicate, repeated twice). ( D ) Representative figures showing antiinflammatory gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. <t>Il10</t> , Arg1 , and Ym1 expression was measured by qPCR ( n = 3 × 3 mice pooled/replicate, repeated twice). A , C , and D were analyzed using a 2-tailed Student’s t test. B was analyzed using 2-way ANOVA followed by Holm-Šídák test for multiple comparisons.
    Gene Exp Il10 Mm00439616 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 542 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gene exp il10 hs00961622 m1
    Macrophages from myeloid-deficient MOF mice demonstrate decreased NF-κB inflammatory genes. ( A ) Representative figures showing Mof deletion in BMDMs and wound macrophages from Lyz2Cre Mof fl/fl mice. BMDMs were derived from Lyz2Cre Mof fl/fl or Mof fl/fl mice. Wound macrophages were sorted by magnetic-activated cell sorting (MACS) from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Mof gene expression was measured by qPCR ( n = 3 × 4 mice pooled/replicate, repeated twice). ( B ) Representative figures showing inflammatory cytokine gene expression in LPS-stimulated BMDMs. BMDMs were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice and treated with LPS or vehicle control for 8 hours. Il1b , Tnf , and Nos2 gene expression was measured by qPCR ( n = 3, repeated twice). ( C ) Representative figures showing baseline inflammatory cytokine gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Il1b ( n = 3), Tnf ( n = 3), and Nos2 ( n = 4) expression was measured by qPCR ( n = 3 mice pooled/replicate, repeated twice). ( D ) Representative figures showing antiinflammatory gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. <t>Il10</t> , Arg1 , and Ym1 expression was measured by qPCR ( n = 3 × 3 mice pooled/replicate, repeated twice). A , C , and D were analyzed using a 2-tailed Student’s t test. B was analyzed using 2-way ANOVA followed by Holm-Šídák test for multiple comparisons.
    Gene Exp Il10 Hs00961622 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 586 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson il 10
    Macrophages from myeloid-deficient MOF mice demonstrate decreased NF-κB inflammatory genes. ( A ) Representative figures showing Mof deletion in BMDMs and wound macrophages from Lyz2Cre Mof fl/fl mice. BMDMs were derived from Lyz2Cre Mof fl/fl or Mof fl/fl mice. Wound macrophages were sorted by magnetic-activated cell sorting (MACS) from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Mof gene expression was measured by qPCR ( n = 3 × 4 mice pooled/replicate, repeated twice). ( B ) Representative figures showing inflammatory cytokine gene expression in LPS-stimulated BMDMs. BMDMs were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice and treated with LPS or vehicle control for 8 hours. Il1b , Tnf , and Nos2 gene expression was measured by qPCR ( n = 3, repeated twice). ( C ) Representative figures showing baseline inflammatory cytokine gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Il1b ( n = 3), Tnf ( n = 3), and Nos2 ( n = 4) expression was measured by qPCR ( n = 3 mice pooled/replicate, repeated twice). ( D ) Representative figures showing antiinflammatory gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. <t>Il10</t> , Arg1 , and Ym1 expression was measured by qPCR ( n = 3 × 3 mice pooled/replicate, repeated twice). A , C , and D were analyzed using a 2-tailed Student’s t test. B was analyzed using 2-way ANOVA followed by Holm-Šídák test for multiple comparisons.
    Il 10, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 4072 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher anti il 10
    Macrophages from myeloid-deficient MOF mice demonstrate decreased NF-κB inflammatory genes. ( A ) Representative figures showing Mof deletion in BMDMs and wound macrophages from Lyz2Cre Mof fl/fl mice. BMDMs were derived from Lyz2Cre Mof fl/fl or Mof fl/fl mice. Wound macrophages were sorted by magnetic-activated cell sorting (MACS) from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Mof gene expression was measured by qPCR ( n = 3 × 4 mice pooled/replicate, repeated twice). ( B ) Representative figures showing inflammatory cytokine gene expression in LPS-stimulated BMDMs. BMDMs were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice and treated with LPS or vehicle control for 8 hours. Il1b , Tnf , and Nos2 gene expression was measured by qPCR ( n = 3, repeated twice). ( C ) Representative figures showing baseline inflammatory cytokine gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Il1b ( n = 3), Tnf ( n = 3), and Nos2 ( n = 4) expression was measured by qPCR ( n = 3 mice pooled/replicate, repeated twice). ( D ) Representative figures showing antiinflammatory gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. <t>Il10</t> , Arg1 , and Ym1 expression was measured by qPCR ( n = 3 × 3 mice pooled/replicate, repeated twice). A , C , and D were analyzed using a 2-tailed Student’s t test. B was analyzed using 2-way ANOVA followed by Holm-Šídák test for multiple comparisons.
    Anti Il 10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 574 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems rhil 10
    PRB782 shows improved biological activity. Induction of <t>IL-10</t> production with treatment of Colo205 cells with induced bacterial supernatant from PRB484 and PRB782 showed PRB782 to have significantly (~15-fold) higher biological activity as compared to PRB484. Data represent the average of 3 biological replicates with error bars indicating standard deviation from the mean. Comparison performed with t -tests. ** P
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    il 10  (Abcam)
    93
    Abcam il 10
    Immunohistochemical expression of anti‐inflammatory cytokines (Magnification: 400×). IL‐4: A, C control placenta (35 wks, 38 wks) showing moderate cytoplasmic expression in ST; B, D, preeclamptic placenta (35 wks, 38 wks) showing mild cytoplasmic expression in ST. <t>IL‐10:</t> E, G, control placenta (35 wks, 38 wks) showing moderate cytoplasmic expression in ST; F, H preeclamptic placenta (32 wks, 38 wks) showing mild cytoplasmic expression in ST
    Il 10, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 231 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gene exp il10 hs00174086 m1
    Immunohistochemical expression of anti‐inflammatory cytokines (Magnification: 400×). IL‐4: A, C control placenta (35 wks, 38 wks) showing moderate cytoplasmic expression in ST; B, D, preeclamptic placenta (35 wks, 38 wks) showing mild cytoplasmic expression in ST. <t>IL‐10:</t> E, G, control placenta (35 wks, 38 wks) showing moderate cytoplasmic expression in ST; F, H preeclamptic placenta (32 wks, 38 wks) showing mild cytoplasmic expression in ST
    Gene Exp Il10 Hs00174086 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 293 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The Jackson Laboratory il 10
    LPS increases protein content of (a) IL-6 and (b) TNFα in gastrocnemius muscles of <t>IL-10</t> +/+ wild-type and IL-10 −/− mice. * p
    Il 10, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 93/100, based on 172 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech il 10
    LPS increases protein content of (a) IL-6 and (b) TNFα in gastrocnemius muscles of <t>IL-10</t> +/+ wild-type and IL-10 −/− mice. * p
    Il 10, supplied by PeproTech, used in various techniques. Bioz Stars score: 93/100, based on 764 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems il10
    Human innate and adaptive immune cells from hNSG mice are functional and respond to cell-specific stimulation. ( A ) Human splenocytes upregulate cell surface expression of activation marker CD69 48 hours after stimulation with human CD3/28 antibody and rhIL2. ( B ) Proportion of hCD4 + and hCD8 + T cells expressing CD69 48 hours after stimulation by hCD3/28 and rhIL2. ( C ) Decrease in CFSE staining demonstrating robust proliferation of human splenocytes stimulated with hCD3/28 and rhIL2. ( D ) Proportion of proliferating splenocytes 96 hours after cell specific stimulation. ( E , F ) Quantification of human interferon gamma (IFNγ) and <t>IL10</t> in culture supernatants after 96 hours of cell specific stimulation. ( G ) Human TNFα quantification in blood serum 1 hour after in vivo injection with 3 mg/kg lipopolysaccharide (LPS). Human IgG ( H ) and IgM ( I ) from blood serum in hNSG mice. Note the absence of human cytokines and antibodies in blood serum of non-engrafted NSG mice treated with LPS, demonstrating species specificity of ELISAs. ND = not detected. Data average ± SEM; n = 2 biological replicates in ( B , D ) n = 4 biological replicates in ( E , F ) n = 3 mice per group in ( G , H ) n = 3 NSG and n = 6 hNSG mice in ( I , J ).
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    Germfree Laboratories Inc germ free il 10
    Secretion of IL-12/23p40 ( A ) and IFN-γ ( B ) by mesenteric lymph node (MLN) cells obtained from WT or <t>IL-10</t> −/− mice fed control or curcumin-supplemented diets and stimulated with cecal antigen extract. MLN cells were stimulated by
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    R&D Systems recombinant il 10
    LPS-induced ROS production is p47 phox dependent in macrophages and ROS scavenger blocks <t>IL-10</t> secretion from HLL mice. A , Representative tracing showed the generation of superoxide as counts per second (CPS) by peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice, stimulated with 100 ng/ml of LPS. B , Peritoneal macrophages from HLL mice were incubated with DPI for 30 min and the cells were left unstimulated or stimulated with LPS (100 ng/ml) for an additional 8 h. IL-10 secretion in the culture supernatant was detected by ELISA. C, E , and F , Peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice were treated with LPS (100 ng/ml) for 8 h. IL-10, IL-6, and TNF-α were measured in the culture supernatant. Results represent mean ± SEM of three separate experiments and *, p
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    Thermo Fisher gene exp il10 rn00563409 m1
    LPS-induced ROS production is p47 phox dependent in macrophages and ROS scavenger blocks <t>IL-10</t> secretion from HLL mice. A , Representative tracing showed the generation of superoxide as counts per second (CPS) by peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice, stimulated with 100 ng/ml of LPS. B , Peritoneal macrophages from HLL mice were incubated with DPI for 30 min and the cells were left unstimulated or stimulated with LPS (100 ng/ml) for an additional 8 h. IL-10 secretion in the culture supernatant was detected by ELISA. C, E , and F , Peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice were treated with LPS (100 ng/ml) for 8 h. IL-10, IL-6, and TNF-α were measured in the culture supernatant. Results represent mean ± SEM of three separate experiments and *, p
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    PeproTech recombinant human interleukin 10
    LPS-induced ROS production is p47 phox dependent in macrophages and ROS scavenger blocks <t>IL-10</t> secretion from HLL mice. A , Representative tracing showed the generation of superoxide as counts per second (CPS) by peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice, stimulated with 100 ng/ml of LPS. B , Peritoneal macrophages from HLL mice were incubated with DPI for 30 min and the cells were left unstimulated or stimulated with LPS (100 ng/ml) for an additional 8 h. IL-10 secretion in the culture supernatant was detected by ELISA. C, E , and F , Peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice were treated with LPS (100 ng/ml) for 8 h. IL-10, IL-6, and TNF-α were measured in the culture supernatant. Results represent mean ± SEM of three separate experiments and *, p
    Recombinant Human Interleukin 10, supplied by PeproTech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology il 10
    Schematic of adaptive immunoregulation of LUT and chlorogenic acid in LPS-induced <t>IL-10</t> expression. CGA enhanced LPS-induced expression of IL-10 and IL-6 and increased the expression of NF-κB, Sp1, C/EBPβ, and δ. The effect of CGA was interfered with LUT by suppressing the phosphorylation of IκB and p38 and downregulated NF-κB activity. In the event, IL-6 was suppressed and IL-10 was not influenced. LPS: Lipopolysaccharide, CGA: Chlorogenic acid, LUT: Luteolin, NF-κB: Nuclear factor-κB, IL-10: Interleukin-10
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    Abcam anti il 10
    Serum levels of <t>IL-10</t> and IL-17 in ulcerative colitis patients and healthy subjects were detected with ELISA. IL-10 and IL-17 levels were significantly higher, compared with control group, ∗ P
    Anti Il 10, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 243 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 243 article reviews
    Price from $9.99 to $1999.99
    anti il 10 - by Bioz Stars, 2020-09
    99/100 stars
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    95
    Thermo Fisher mouse il 10
    The effect of amino acid substitution at the RRCHR region of mouse <t>IL-10</t> on biological activities. (A) The secondary structure of IL-10 mutants by CD spectroscopy adopted from unpublished data ( 36 ). (B) The ED50 of natural IL-10 was calculated as flow: Nm RRCHR (un-mutant) (black line) (0.1 ng/mL), Nm RACHR (green line) (0.28 ng/mL), Nm AACHR (blue line) (0.96 ng/mL), and Nm ARCHA (red line) (59.28 ng/mL). (C) The activation of cellular signaling (p-STAT3) determines by western blot after lysate from spleen cells of the C57BL/6 mouse treated Nm mutants and un-mutant (control) in dose-response and using the total STAT3 as an internal control. (D) The ED50 of stable IL-10 was calculated as flow: STm RRCHR (un-mutant) (black line) (0.36 ng/mL), STm RACHR (green line) (1.6 ng/mL), STm AACHR (blue line) (2.55 ng/mL), and ST ARCHA (red line) (3.02 ng/mL). (E) The activation p-STAT3 determines by western blot from spleen cells lysate of the C57BL/6 mouse treated STm mutants and un-mutant (control) in dose-response and using the total STAT3 as an internal control. The data represented as AUC (Area Under the Curve). All data are representative of two independent experiments, with triplicate cultures per experiment ( N = 2, n = 3), and bars represent standard error of the mean.
    Mouse Il 10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 180 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse il 10/product/Thermo Fisher
    Average 95 stars, based on 180 article reviews
    Price from $9.99 to $1999.99
    mouse il 10 - by Bioz Stars, 2020-09
    95/100 stars
      Buy from Supplier

    Image Search Results


    IL-10 expression supports EMT in invasive LSCC. (a) IL-10 plasmatic concentrations by ELISA in healthy donors (HD) ( n = 9) and LSCC patients ( n = 20), and Mann–Whitney U test performed for statistical analysis; data are shown as mean ± SD, ∗ p ≤ 0.05. (b) Representative IL-10 and CD68 stainings in laryngeal tissues (magnification ×40) (yellow arrow: normal epithelium, black arrow: tumor cells, and green arrow: stomal cells). (c) Percentage of stromal cells producing IL-10 was evaluated by IHC and correlated to the percentage of CD68 + stromal macrophage by the Pearson correlation test. (d) Correlogram: EMT biomarkers expression levels versus IL-10 and NOS2 expression in invasive LSCC ( n = 20).

    Journal: Journal of Oncology

    Article Title: Expression Analysis of the Mediators of Epithelial to Mesenchymal Transition and Early Risk Assessment of Therapeutic Failure in Laryngeal Carcinoma

    doi: 10.1155/2019/5649846

    Figure Lengend Snippet: IL-10 expression supports EMT in invasive LSCC. (a) IL-10 plasmatic concentrations by ELISA in healthy donors (HD) ( n = 9) and LSCC patients ( n = 20), and Mann–Whitney U test performed for statistical analysis; data are shown as mean ± SD, ∗ p ≤ 0.05. (b) Representative IL-10 and CD68 stainings in laryngeal tissues (magnification ×40) (yellow arrow: normal epithelium, black arrow: tumor cells, and green arrow: stomal cells). (c) Percentage of stromal cells producing IL-10 was evaluated by IHC and correlated to the percentage of CD68 + stromal macrophage by the Pearson correlation test. (d) Correlogram: EMT biomarkers expression levels versus IL-10 and NOS2 expression in invasive LSCC ( n = 20).

    Article Snippet: Immunohistochemistry (IHC) Tissue sections of 5 μ m were tested against E-cadherin (NCH-38, prediluted format, Dako), β -catenin (17C2, prediluted format, Novocastra), vimentin (V9, 1/500 Novocastra), NOS2 (SAB4502012, 1/1000, Sigma), NF-κ B (p65, 2A12A7, 1/500, Invitrogen), TGF-β (TB21, 1/1000, Thermofisher), IL-6 (10C12, 1/100, Novocastra), IL-17 (50104, 1/100, Invitrogen), IL-10 (945A2A5, 1/100, Invitrogen), CD68 (KP1, prediluted format, Dako), and MMP-9 (2C3, 1/200, Santa Cruz Biotechnology).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Immunohistochemistry

    KM survival analysis for LSCC patients ( n = 20) per clusters and analyzed EMT inducers molecules. Patients' stratification was based on the hierarchical clustering and the median concentrations of the tested biomarkers as cutoffs: IL-17A (42 pg/ml), TGF- β (543.5 pg/ml), IL-6 (34.86 pg/ml), IL-10 (10.5 pg/ml), NO (32.69 μ M). p values were calculated using the log-rank test.

    Journal: Journal of Oncology

    Article Title: Expression Analysis of the Mediators of Epithelial to Mesenchymal Transition and Early Risk Assessment of Therapeutic Failure in Laryngeal Carcinoma

    doi: 10.1155/2019/5649846

    Figure Lengend Snippet: KM survival analysis for LSCC patients ( n = 20) per clusters and analyzed EMT inducers molecules. Patients' stratification was based on the hierarchical clustering and the median concentrations of the tested biomarkers as cutoffs: IL-17A (42 pg/ml), TGF- β (543.5 pg/ml), IL-6 (34.86 pg/ml), IL-10 (10.5 pg/ml), NO (32.69 μ M). p values were calculated using the log-rank test.

    Article Snippet: Immunohistochemistry (IHC) Tissue sections of 5 μ m were tested against E-cadherin (NCH-38, prediluted format, Dako), β -catenin (17C2, prediluted format, Novocastra), vimentin (V9, 1/500 Novocastra), NOS2 (SAB4502012, 1/1000, Sigma), NF-κ B (p65, 2A12A7, 1/500, Invitrogen), TGF-β (TB21, 1/1000, Thermofisher), IL-6 (10C12, 1/100, Novocastra), IL-17 (50104, 1/100, Invitrogen), IL-10 (945A2A5, 1/100, Invitrogen), CD68 (KP1, prediluted format, Dako), and MMP-9 (2C3, 1/200, Santa Cruz Biotechnology).

    Techniques:

    4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) recovered high-fat diet (HFD)-impaired insulin signaling and inflammation in visceral adipose (VAD) tissue of LDLR −/− mice. ( A ) The peripheral-tissue insulin signaling pathway [insulin receptor, pyruvate dehydrogenase kinase isozyme 1 (PDK1), Akt, glycogen synthase kinase 3β (GSK3β)] was analyzed by western immunoblots. ( B ) Quantification of insulin receptor-α and -β expression, measured by imageQuant. Amounts of phosphorylated and total PDK1 ( C ), Akt ( D ), and GSK3β ( E ). ( F ) VAD tissue inflammatory cytokines and macrophage markers [interleukin (IL)-6, IL-10, CD11b, F4/80] were analyzed by western immunoblots, with protein levels quantified by imageQuant. ( G ) Tumor necrosis factor-α (TNF-α) expression in VAD tissue was analyzed by ELISA. ( H ) Serine protease activity in VAD tissue of mice receiving the chow, HFD, and HFD + AEBSF treatments ( n = 6/group). * P

    Journal: Scientific Reports

    Article Title: Inhibition of Serine Protease Activity Protects Against High Fat Diet-Induced Inflammation and Insulin Resistance

    doi: 10.1038/s41598-020-58361-4

    Figure Lengend Snippet: 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) recovered high-fat diet (HFD)-impaired insulin signaling and inflammation in visceral adipose (VAD) tissue of LDLR −/− mice. ( A ) The peripheral-tissue insulin signaling pathway [insulin receptor, pyruvate dehydrogenase kinase isozyme 1 (PDK1), Akt, glycogen synthase kinase 3β (GSK3β)] was analyzed by western immunoblots. ( B ) Quantification of insulin receptor-α and -β expression, measured by imageQuant. Amounts of phosphorylated and total PDK1 ( C ), Akt ( D ), and GSK3β ( E ). ( F ) VAD tissue inflammatory cytokines and macrophage markers [interleukin (IL)-6, IL-10, CD11b, F4/80] were analyzed by western immunoblots, with protein levels quantified by imageQuant. ( G ) Tumor necrosis factor-α (TNF-α) expression in VAD tissue was analyzed by ELISA. ( H ) Serine protease activity in VAD tissue of mice receiving the chow, HFD, and HFD + AEBSF treatments ( n = 6/group). * P

    Article Snippet: Antibodies against mouse interleukin (IL)-6 (MAB-406) and IL-10 (AF-519) and all proteins were detected using horseradish peroxidase (HRP)-conjugated secondary antibodies (R & D Systems, Minneapolis, MN).

    Techniques: Mouse Assay, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Activity Assay

    Construction and validation of Lenti-IL-27HA and Lenti-IL-35HA. ( A , B ). Graphic representation of the lentiviral constructs used to express IL-27 ( A ) and IL-35 ( B ). ( C , D ) Cytokines production was assessed by ELISA (IL-27, C ) or WB (IL-35, D ). ( E – G ). Affinity purified IL-27 (white bars) and IL-35 (black bars) were used to treat polarized CD4 + T cells and measure mRNA expression of IL-2 ( E ), IL-17 ( F ), gm-csf ( G ), and IL-10 ( F ) (mean ± sd). NC = no cytokines. *p

    Journal: Scientific Reports

    Article Title: IL-27, but not IL-35, inhibits neuroinflammation through modulating GM-CSF expression

    doi: 10.1038/s41598-017-16702-w

    Figure Lengend Snippet: Construction and validation of Lenti-IL-27HA and Lenti-IL-35HA. ( A , B ). Graphic representation of the lentiviral constructs used to express IL-27 ( A ) and IL-35 ( B ). ( C , D ) Cytokines production was assessed by ELISA (IL-27, C ) or WB (IL-35, D ). ( E – G ). Affinity purified IL-27 (white bars) and IL-35 (black bars) were used to treat polarized CD4 + T cells and measure mRNA expression of IL-2 ( E ), IL-17 ( F ), gm-csf ( G ), and IL-10 ( F ) (mean ± sd). NC = no cytokines. *p

    Article Snippet: Arg-1 (Mm00475988_m1); inos (Mm00440502_m1); ym1 (Mm00657889_mH); cd274 (Mm00452054_m1); tbx21 (Mm00450960_m1); csf2 (Mm01290062_m1); IL-17a (Mm00439618_m1); rorc (Mm00441144_g1); foxp3 (Mm00475156_m1); IL-2 (Mm00434256_m1); IL-10 (Mm00439614_m1) and gapdh (4352339E) mRNA levels were measured by real-time RT-PCR (Applied Biosystem, Invitrogen).

    Techniques: Construct, Enzyme-linked Immunosorbent Assay, Western Blot, Affinity Purification, Expressing

    IL-10–producing Tfh cells emerge after vaccination, and IL-10, but neither T regs nor Tfr cells, limits Tfh-dependent vaccine responses in aged mice. ( A ) Plots show IA b -NP tetramer staining in immunized and unimmunized young (3.5 months, n = 5) and aged (18 months, n = 5) IL-10GFP–FOXP3RFP dual-reporter mice. Graphs show the frequency and number of NP-specific FoxP3 − CD4 + T cells that are IL-10 + and the number of NP-specific FoxP3 − IL-10 + T cells that are CXCR5 + PD1 + (means ± SEM). ( B ) Aged (17 months, n ≥ 7) WT C57BL/6 (isotype or anti–IL-10) and (20 months, n = 5) FoxP3 − DTR [phosphate-buffered saline (PBS)– or DT-treated] mice were immunized with nitrophenol-keyhole limpet hemocyanin (NP-KLH) in alum. Plots display the frequency of germinal center (GC) B cells (NP-specific) gated on Fas + GL7 + . Graphs show the frequency and number of B cells that are IgG1 + NP + (means ± SEM) and serum levels of immunoglobulin (Ig) specific for NP (IgG1) (means ± SEM). Data are pooled from two independent experiments. ( C to E ) Tfh10 cells accumulate during aging in humans. Human spleen cells from young (Y) (median, 18.8; range, 18 to 26 years; three males and five females) and old (O) (median, 62; range, 60 to 67 years; four males and four females) individuals were analyzed by flow cytometry. (C) The frequencies of CD3 + CD4 + CD45RO + FoxP3 − that are CXCR5 + PD1 + (means ± SEM). (D and E) CD4 + T cells were bead-purified by negative selection; FACS-sorted memory CD4 + T cells (CD45RO + ) into Tfh cells (CD25 − CD127 + PD1 + CXCR5 + ), T regs (CD25 + CD127 − PD1 − CXCR5 − ), and other memory cells (CD25 − CD127 + PD1 − CXCR5 − ) were either stimulated in vitro with anti-CD3/CD28 beads or unstimulated, and cytokines were analyzed by Luminex (means ± SEM). Each individual is represented by a symbol. * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001, Student’s t test. BSS, balanced salt solution.

    Journal: Science Advances

    Article Title: IL-10–producing Tfh cells accumulate with age and link inflammation with age-related immune suppression

    doi: 10.1126/sciadv.abb0806

    Figure Lengend Snippet: IL-10–producing Tfh cells emerge after vaccination, and IL-10, but neither T regs nor Tfr cells, limits Tfh-dependent vaccine responses in aged mice. ( A ) Plots show IA b -NP tetramer staining in immunized and unimmunized young (3.5 months, n = 5) and aged (18 months, n = 5) IL-10GFP–FOXP3RFP dual-reporter mice. Graphs show the frequency and number of NP-specific FoxP3 − CD4 + T cells that are IL-10 + and the number of NP-specific FoxP3 − IL-10 + T cells that are CXCR5 + PD1 + (means ± SEM). ( B ) Aged (17 months, n ≥ 7) WT C57BL/6 (isotype or anti–IL-10) and (20 months, n = 5) FoxP3 − DTR [phosphate-buffered saline (PBS)– or DT-treated] mice were immunized with nitrophenol-keyhole limpet hemocyanin (NP-KLH) in alum. Plots display the frequency of germinal center (GC) B cells (NP-specific) gated on Fas + GL7 + . Graphs show the frequency and number of B cells that are IgG1 + NP + (means ± SEM) and serum levels of immunoglobulin (Ig) specific for NP (IgG1) (means ± SEM). Data are pooled from two independent experiments. ( C to E ) Tfh10 cells accumulate during aging in humans. Human spleen cells from young (Y) (median, 18.8; range, 18 to 26 years; three males and five females) and old (O) (median, 62; range, 60 to 67 years; four males and four females) individuals were analyzed by flow cytometry. (C) The frequencies of CD3 + CD4 + CD45RO + FoxP3 − that are CXCR5 + PD1 + (means ± SEM). (D and E) CD4 + T cells were bead-purified by negative selection; FACS-sorted memory CD4 + T cells (CD45RO + ) into Tfh cells (CD25 − CD127 + PD1 + CXCR5 + ), T regs (CD25 + CD127 − PD1 − CXCR5 − ), and other memory cells (CD25 − CD127 + PD1 − CXCR5 − ) were either stimulated in vitro with anti-CD3/CD28 beads or unstimulated, and cytokines were analyzed by Luminex (means ± SEM). Each individual is represented by a symbol. * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001, Student’s t test. BSS, balanced salt solution.

    Article Snippet: A luminescent ELISA was performed using anti–IL-6 (MP5-20F3, Invitrogen) or anti–IL-10 (JES5-2A5, BD Biosciences) as the coating antibody.

    Techniques: Mouse Assay, Staining, Flow Cytometry, Purification, Selection, FACS, In Vitro, Luminex

    Aged mice have increased systemic levels of IL-10. ( A ) Serum IL-10 (means ± SEM) levels in young (4 months, n = 6) and aged (18 months, n = 5) mice, representative of four independent experiments. ( B ) Mean fold change in IL-10 mRNA gene expression (means ± SEM) from the spleen, liver, gut, lymph nodes (LNs), inguinal white adipose tissue (iWAT), epididymal white adipose tissue (eWAT), and brown adipose tissue (BAT) from individual young (2 months, n = 4 to 8) and aged (21 months, n = 5 to 9) C57BL/6 mice. Dashed line represents equal aged:young ratio. Data pooled from two independent experiments. ( C ) Splenocytes from young (1.5 months, n = 3) and aged (18 months, n = 5) IL-10 gfp (VertX) mice were analyzed by flow cytometry. Upper graph shows the frequency of cells that are green fluorescent protein–positive (GFP + ) (means ± SEM). Lower graph shows the average level of GFP expression in aged CD4 + , CD8 + , CD19 + , and CD19 − that are GFP + (means ± SEM). ( D ) IL-10 levels (means ± SEM) from phorbol 12-myristate 13-acetate and ionomycin (P + I)–stimulated cells sorted from young (3.5 months, n = 4) and aged (24 months, n = 4) FoxP3–internal ribosomal entry site (IRES)–diphtheria toxin receptor (DTR)–GFP mice. ( E ) Gating strategy, frequencies, and numbers of FoxP3 + or FoxP3 − that are IL-10 + from P + I–stimulated splenocytes from young (1.5 months, n = 4) and aged (23 months, n = 4) C57BL/6 mice (means ± SEM). ( F ) Serum IL-10 levels (means ± SEM) in young (2.5 months, n = 6) and aged (18 months, n = 14) C57BL/6 mice treated with anti-CD4 or isotype control or DT-treated FoxP3-DTR mice (19 months, n = 6). Data are pooled from two independent experiments. ( G ) Percentage of FoxP3 − splenocytes that are IL-10 + (means ± SEM) from aged (18 months, n = 8) and DT-treated FoxP3-DTR (19 months, n = 6) mice. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001, Student’s t test. MFI, mean fluorescence intensity.

    Journal: Science Advances

    Article Title: IL-10–producing Tfh cells accumulate with age and link inflammation with age-related immune suppression

    doi: 10.1126/sciadv.abb0806

    Figure Lengend Snippet: Aged mice have increased systemic levels of IL-10. ( A ) Serum IL-10 (means ± SEM) levels in young (4 months, n = 6) and aged (18 months, n = 5) mice, representative of four independent experiments. ( B ) Mean fold change in IL-10 mRNA gene expression (means ± SEM) from the spleen, liver, gut, lymph nodes (LNs), inguinal white adipose tissue (iWAT), epididymal white adipose tissue (eWAT), and brown adipose tissue (BAT) from individual young (2 months, n = 4 to 8) and aged (21 months, n = 5 to 9) C57BL/6 mice. Dashed line represents equal aged:young ratio. Data pooled from two independent experiments. ( C ) Splenocytes from young (1.5 months, n = 3) and aged (18 months, n = 5) IL-10 gfp (VertX) mice were analyzed by flow cytometry. Upper graph shows the frequency of cells that are green fluorescent protein–positive (GFP + ) (means ± SEM). Lower graph shows the average level of GFP expression in aged CD4 + , CD8 + , CD19 + , and CD19 − that are GFP + (means ± SEM). ( D ) IL-10 levels (means ± SEM) from phorbol 12-myristate 13-acetate and ionomycin (P + I)–stimulated cells sorted from young (3.5 months, n = 4) and aged (24 months, n = 4) FoxP3–internal ribosomal entry site (IRES)–diphtheria toxin receptor (DTR)–GFP mice. ( E ) Gating strategy, frequencies, and numbers of FoxP3 + or FoxP3 − that are IL-10 + from P + I–stimulated splenocytes from young (1.5 months, n = 4) and aged (23 months, n = 4) C57BL/6 mice (means ± SEM). ( F ) Serum IL-10 levels (means ± SEM) in young (2.5 months, n = 6) and aged (18 months, n = 14) C57BL/6 mice treated with anti-CD4 or isotype control or DT-treated FoxP3-DTR mice (19 months, n = 6). Data are pooled from two independent experiments. ( G ) Percentage of FoxP3 − splenocytes that are IL-10 + (means ± SEM) from aged (18 months, n = 8) and DT-treated FoxP3-DTR (19 months, n = 6) mice. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001, Student’s t test. MFI, mean fluorescence intensity.

    Article Snippet: A luminescent ELISA was performed using anti–IL-6 (MP5-20F3, Invitrogen) or anti–IL-10 (JES5-2A5, BD Biosciences) as the coating antibody.

    Techniques: Mouse Assay, Expressing, Flow Cytometry, Fluorescence

    IL-10–producing FoxP3 − CD4 + T cells in aged mice are predominantly Tfh cells. ( A ) Splenocytes from young (2 months, n = 6) and aged (18 months, n = 6) C57BL/6 mice were stimulated with P + I, stained with antibodies against TCRβ, CD8, FoxP3, IL-10, and IL-21, and analyzed by flow cytometry. The representative plots and graphs show the frequencies and total numbers of IL-21 + cells originating from FoxP3 − IL10 + cells (means ± SEM). Data are representative of at least two independent experiments. ( B ) Splenocytes from young (2 months, n = 4) and aged (18 months, n = 4) C57BL/6 mice were stimulated as above and stained with antibodies against TCRβ, CD8, FoxP3, CXCR5, PD1, and IL-10 and analyzed by flow cytometry. The representative plots and graphs show the frequencies and total numbers of CXCR5 + PD1 + cells originating from FoxP3 − IL10 + cells (means ± SEM). * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001, Student’s t test.

    Journal: Science Advances

    Article Title: IL-10–producing Tfh cells accumulate with age and link inflammation with age-related immune suppression

    doi: 10.1126/sciadv.abb0806

    Figure Lengend Snippet: IL-10–producing FoxP3 − CD4 + T cells in aged mice are predominantly Tfh cells. ( A ) Splenocytes from young (2 months, n = 6) and aged (18 months, n = 6) C57BL/6 mice were stimulated with P + I, stained with antibodies against TCRβ, CD8, FoxP3, IL-10, and IL-21, and analyzed by flow cytometry. The representative plots and graphs show the frequencies and total numbers of IL-21 + cells originating from FoxP3 − IL10 + cells (means ± SEM). Data are representative of at least two independent experiments. ( B ) Splenocytes from young (2 months, n = 4) and aged (18 months, n = 4) C57BL/6 mice were stimulated as above and stained with antibodies against TCRβ, CD8, FoxP3, CXCR5, PD1, and IL-10 and analyzed by flow cytometry. The representative plots and graphs show the frequencies and total numbers of CXCR5 + PD1 + cells originating from FoxP3 − IL10 + cells (means ± SEM). * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001, Student’s t test.

    Article Snippet: A luminescent ELISA was performed using anti–IL-6 (MP5-20F3, Invitrogen) or anti–IL-10 (JES5-2A5, BD Biosciences) as the coating antibody.

    Techniques: Mouse Assay, Staining, Flow Cytometry

    IL-6 is required for Tfh10 cells and for elevated levels of IL-10 in aged mice. ( A and B ) Splenocytes from young (2 months, n ≥ 4 per group) and aged (17 months, n ≥ 4 per group) C57BL/6 or IL-6 −/− mice were stimulated with P + I, stained with antibody against TCRβ, CD8, CXCR5, PD1, FoxP3, and IL-10 and analyzed by flow cytometry. The representative plots and bar graphs show the frequency and total number of FoxP3 − that are (A) CXCR5 + PD1 + and (B) those that produce IL-10 (means ± SEM). ( C ) Aged C57BL/6 (17 months, n = 9) and IL-6 −/− (17 months, n = 9) mice were intravenously injected with biotinylated anti–IL-10 antibodies, serum was collected 24 hours later, and IL-10 levels were measured by ELISA. Graph shows the average serum IL-10 (means ± SEM). ( D ) Aged C57BL/6 mice were treated with isotype control (19 months, n = 6) or α–IL-6 blocking antibody (19 months, n = 8) on day 0 and euthanized on day 2. Splenocytes were stimulated with P + I, stained with antibody against TCRβ, CD8, PD1, FoxP3, and IL-10, and analyzed by flow cytometry. The representative bar graph shows the frequency of FoxP3 − that are IL-10 + (means ± SEM). * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001, Student’s t test.

    Journal: Science Advances

    Article Title: IL-10–producing Tfh cells accumulate with age and link inflammation with age-related immune suppression

    doi: 10.1126/sciadv.abb0806

    Figure Lengend Snippet: IL-6 is required for Tfh10 cells and for elevated levels of IL-10 in aged mice. ( A and B ) Splenocytes from young (2 months, n ≥ 4 per group) and aged (17 months, n ≥ 4 per group) C57BL/6 or IL-6 −/− mice were stimulated with P + I, stained with antibody against TCRβ, CD8, CXCR5, PD1, FoxP3, and IL-10 and analyzed by flow cytometry. The representative plots and bar graphs show the frequency and total number of FoxP3 − that are (A) CXCR5 + PD1 + and (B) those that produce IL-10 (means ± SEM). ( C ) Aged C57BL/6 (17 months, n = 9) and IL-6 −/− (17 months, n = 9) mice were intravenously injected with biotinylated anti–IL-10 antibodies, serum was collected 24 hours later, and IL-10 levels were measured by ELISA. Graph shows the average serum IL-10 (means ± SEM). ( D ) Aged C57BL/6 mice were treated with isotype control (19 months, n = 6) or α–IL-6 blocking antibody (19 months, n = 8) on day 0 and euthanized on day 2. Splenocytes were stimulated with P + I, stained with antibody against TCRβ, CD8, PD1, FoxP3, and IL-10, and analyzed by flow cytometry. The representative bar graph shows the frequency of FoxP3 − that are IL-10 + (means ± SEM). * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001, Student’s t test.

    Article Snippet: A luminescent ELISA was performed using anti–IL-6 (MP5-20F3, Invitrogen) or anti–IL-10 (JES5-2A5, BD Biosciences) as the coating antibody.

    Techniques: Mouse Assay, Staining, Flow Cytometry, Injection, Enzyme-linked Immunosorbent Assay, Blocking Assay

    IL-21 contributes to accrual of Tfh10 cells and regulates the systemic IL-6/IL-10 balance. ( A ) Splenocytes from young (2.5 months, n ≥ 4 per group) and aged (16 months, n ≥ 4 per group) C57BL/6 or IL-6 −/− mice were stimulated with P + I, stained with antibody against TCRβ, CD8, FoxP3, and IL-21, and analyzed by flow cytometry. Bar graphs show the frequency and total number of FoxP3 − that are IL-21 + (means ± SEM). ( B and C ) Splenocytes from young (2 months, n ≥ 4 per group) or aged (17 months, n ≥ 3 per group) C57BL/6 or IL-21 −/− mice were stimulated as above, stained with antibody against TCRβ, CD8, CXCR5, PD1, FoxP3, and IL-10, and analyzed by flow cytometry. The representative plots and bar graphs show the frequency and total number of FoxP3 − cells that are (B) CXCR5 + PD1 + and (C) those that produce IL-10 (means ± SEM). Data are pooled from two independent experiments. ( D and E ) Aged C57BL/6 (17 months, n = 4) and IL-21 −/− (17 months, n = 3) mice were intravenously injected with biotinylated anti–IL-10 and anti–IL-6 capturing antibodies, serum was collected 24 hours later, and IL-10 and IL-6 levels were measured by ELISA. Graphs show the average serum (D) IL-10 and (E) IL-6 (means ± SEM). * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001, Student’s t test.

    Journal: Science Advances

    Article Title: IL-10–producing Tfh cells accumulate with age and link inflammation with age-related immune suppression

    doi: 10.1126/sciadv.abb0806

    Figure Lengend Snippet: IL-21 contributes to accrual of Tfh10 cells and regulates the systemic IL-6/IL-10 balance. ( A ) Splenocytes from young (2.5 months, n ≥ 4 per group) and aged (16 months, n ≥ 4 per group) C57BL/6 or IL-6 −/− mice were stimulated with P + I, stained with antibody against TCRβ, CD8, FoxP3, and IL-21, and analyzed by flow cytometry. Bar graphs show the frequency and total number of FoxP3 − that are IL-21 + (means ± SEM). ( B and C ) Splenocytes from young (2 months, n ≥ 4 per group) or aged (17 months, n ≥ 3 per group) C57BL/6 or IL-21 −/− mice were stimulated as above, stained with antibody against TCRβ, CD8, CXCR5, PD1, FoxP3, and IL-10, and analyzed by flow cytometry. The representative plots and bar graphs show the frequency and total number of FoxP3 − cells that are (B) CXCR5 + PD1 + and (C) those that produce IL-10 (means ± SEM). Data are pooled from two independent experiments. ( D and E ) Aged C57BL/6 (17 months, n = 4) and IL-21 −/− (17 months, n = 3) mice were intravenously injected with biotinylated anti–IL-10 and anti–IL-6 capturing antibodies, serum was collected 24 hours later, and IL-10 and IL-6 levels were measured by ELISA. Graphs show the average serum (D) IL-10 and (E) IL-6 (means ± SEM). * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001, Student’s t test.

    Article Snippet: A luminescent ELISA was performed using anti–IL-6 (MP5-20F3, Invitrogen) or anti–IL-10 (JES5-2A5, BD Biosciences) as the coating antibody.

    Techniques: Mouse Assay, Staining, Flow Cytometry, Injection, Enzyme-linked Immunosorbent Assay

    IL-21–driven repression of Bim in aged Tfh10 cells results in their enhanced survival. Splenocytes from young (1.5 months, n = 4) and aged (18 months, n = 4) mice were stimulated with P + I, stained, and analyzed by flow cytometry. ( A ) Graph shows the frequency of FoxP3 − IL-10 + cells that are Ki67 + (means ± SEM). ( B ) Spleen cells were pooled from young (2 months, n = 9) and aged (18 months, n = 6) IL-10 gfp (VertX) mice, and CXCR5 + PD1 + GFP + cells were fluorescence-activated cell sorting (FACS)–sorted and cultured. After 15 hours in culture, cells were stained for FoxP3 and a viability dye. Graph shows the percentage of live cells in gated FoxP3 − cells (means ± SEM). ( C ) Splenocytes from young (2 months, n = 4) and aged (18 months, n = 4) mice were stimulated as mentioned in (A). Graph shows the level of expression of Bim in FoxP3 − IL-10 + cells (means ± SEM). ( D ) Splenocytes from 6-month-old WT C57BL/6 and Bim −/− mice ( n = 6 per group) were stimulated as mentioned in (A), stained, and analyzed by flow cytometry. Plots and bar graphs show the frequency and total number of FoxP3 − that are IL-10 + (means ± SEM). ( E ) Splenocytes from aged (17 months, n = 3 to 4 per group) C57BL/6 or IL-21 −/− mice were stimulated as mentioned in (A), stained, and analyzed by flow cytometry. Graph shows the levels of expression of Bim in FoxP3 − CXCR5 + PD1 + that are IL-10 + cells (means ± SEM). * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001, Student’s t test.

    Journal: Science Advances

    Article Title: IL-10–producing Tfh cells accumulate with age and link inflammation with age-related immune suppression

    doi: 10.1126/sciadv.abb0806

    Figure Lengend Snippet: IL-21–driven repression of Bim in aged Tfh10 cells results in their enhanced survival. Splenocytes from young (1.5 months, n = 4) and aged (18 months, n = 4) mice were stimulated with P + I, stained, and analyzed by flow cytometry. ( A ) Graph shows the frequency of FoxP3 − IL-10 + cells that are Ki67 + (means ± SEM). ( B ) Spleen cells were pooled from young (2 months, n = 9) and aged (18 months, n = 6) IL-10 gfp (VertX) mice, and CXCR5 + PD1 + GFP + cells were fluorescence-activated cell sorting (FACS)–sorted and cultured. After 15 hours in culture, cells were stained for FoxP3 and a viability dye. Graph shows the percentage of live cells in gated FoxP3 − cells (means ± SEM). ( C ) Splenocytes from young (2 months, n = 4) and aged (18 months, n = 4) mice were stimulated as mentioned in (A). Graph shows the level of expression of Bim in FoxP3 − IL-10 + cells (means ± SEM). ( D ) Splenocytes from 6-month-old WT C57BL/6 and Bim −/− mice ( n = 6 per group) were stimulated as mentioned in (A), stained, and analyzed by flow cytometry. Plots and bar graphs show the frequency and total number of FoxP3 − that are IL-10 + (means ± SEM). ( E ) Splenocytes from aged (17 months, n = 3 to 4 per group) C57BL/6 or IL-21 −/− mice were stimulated as mentioned in (A), stained, and analyzed by flow cytometry. Graph shows the levels of expression of Bim in FoxP3 − CXCR5 + PD1 + that are IL-10 + cells (means ± SEM). * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001, Student’s t test.

    Article Snippet: A luminescent ELISA was performed using anti–IL-6 (MP5-20F3, Invitrogen) or anti–IL-10 (JES5-2A5, BD Biosciences) as the coating antibody.

    Techniques: Mouse Assay, Staining, Flow Cytometry, Fluorescence, FACS, Cell Culture, Expressing

    Neutralization of IL-10 increases IFN-γ, TNF-α and IL-6 production in response to B. pseudomallei. PBMCs of one representative donor were stimulated with killed B. pseudomallei at various ratios (0.3:1, 3:1 or 30:1) with or without the addition of anti-IL-10 mAb for 48 hours in vitro ( a ). Kinetics of PMBCs cytokine responses to killed B. pseudomallei (30:1 ratio) in the presence or absence of anti-IL-10 mAb for 6, 12, 24 or 48 hours in vitro ( b ). IFN-γ, TNF-α and IL-6 in cell supernatants were measured by ELISA. The data are presented with the mean and the standard deviation.

    Journal: Scientific Reports

    Article Title: Interleukin 10 inhibits pro-inflammatory cytokine responses and killing of Burkholderia pseudomallei

    doi: 10.1038/srep42791

    Figure Lengend Snippet: Neutralization of IL-10 increases IFN-γ, TNF-α and IL-6 production in response to B. pseudomallei. PBMCs of one representative donor were stimulated with killed B. pseudomallei at various ratios (0.3:1, 3:1 or 30:1) with or without the addition of anti-IL-10 mAb for 48 hours in vitro ( a ). Kinetics of PMBCs cytokine responses to killed B. pseudomallei (30:1 ratio) in the presence or absence of anti-IL-10 mAb for 6, 12, 24 or 48 hours in vitro ( b ). IFN-γ, TNF-α and IL-6 in cell supernatants were measured by ELISA. The data are presented with the mean and the standard deviation.

    Article Snippet: Intracellular cytokine production was detected with anti-IFN-γ FITC (clone 4SB3), anti-TNF-α PE (clone MAb11) (BioLegend) and anti-IL-10 PE (clone JES3-9D7) (BD Biosciences).

    Techniques: Neutralization, In Vitro, Enzyme-linked Immunosorbent Assay, Standard Deviation

    CD3 − CD14 + monocytes are the main IL-10 producers in response to B. pseudomallei . PBMCs from healthy seropositive individuals were incubated with killed B. pseudomallei for 20 hours and, intracellular IL-10 versus cell surface expression of either CD3 or CD14 detected by flow cytometry. The profile ( a ) of one representative donor was gated on monocytes ( b ) and lymphocytes ( c ). d and e shows the frequency of IL-10 producing CD3 − CD14 + monocytes ( d ) or CD3 + lymphocytes ( e ) from multiple individuals (n = 4). The data in ( d ) are presented with the mean and the standard deviation. Statistical significance of killed B. pseudomallei versus medium alone was determined using paired T test; ns, non significant, * p

    Journal: Scientific Reports

    Article Title: Interleukin 10 inhibits pro-inflammatory cytokine responses and killing of Burkholderia pseudomallei

    doi: 10.1038/srep42791

    Figure Lengend Snippet: CD3 − CD14 + monocytes are the main IL-10 producers in response to B. pseudomallei . PBMCs from healthy seropositive individuals were incubated with killed B. pseudomallei for 20 hours and, intracellular IL-10 versus cell surface expression of either CD3 or CD14 detected by flow cytometry. The profile ( a ) of one representative donor was gated on monocytes ( b ) and lymphocytes ( c ). d and e shows the frequency of IL-10 producing CD3 − CD14 + monocytes ( d ) or CD3 + lymphocytes ( e ) from multiple individuals (n = 4). The data in ( d ) are presented with the mean and the standard deviation. Statistical significance of killed B. pseudomallei versus medium alone was determined using paired T test; ns, non significant, * p

    Article Snippet: Intracellular cytokine production was detected with anti-IFN-γ FITC (clone 4SB3), anti-TNF-α PE (clone MAb11) (BioLegend) and anti-IL-10 PE (clone JES3-9D7) (BD Biosciences).

    Techniques: Incubation, Expressing, Flow Cytometry, Cytometry, Standard Deviation

    IFN-γ production in response to B. pseudomallei is reduced in individuals with DM. Whole blood of healthy seropositive individuals (n = 75) and individuals with DM (n = 11) were incubated with killed B. pseudomallei (30:1 ratio), 10 μg/ml of E. coli LPS or medium alone and culture supernatants assayed for IFN-γ ( a ) and IL-10 ( b ) by ELISA after 48 hours in vitro . The data are presented with the mean and the standard deviation. Statistical significance was determined by Mann-Whitney test; ns, non significant, * p

    Journal: Scientific Reports

    Article Title: Interleukin 10 inhibits pro-inflammatory cytokine responses and killing of Burkholderia pseudomallei

    doi: 10.1038/srep42791

    Figure Lengend Snippet: IFN-γ production in response to B. pseudomallei is reduced in individuals with DM. Whole blood of healthy seropositive individuals (n = 75) and individuals with DM (n = 11) were incubated with killed B. pseudomallei (30:1 ratio), 10 μg/ml of E. coli LPS or medium alone and culture supernatants assayed for IFN-γ ( a ) and IL-10 ( b ) by ELISA after 48 hours in vitro . The data are presented with the mean and the standard deviation. Statistical significance was determined by Mann-Whitney test; ns, non significant, * p

    Article Snippet: Intracellular cytokine production was detected with anti-IFN-γ FITC (clone 4SB3), anti-TNF-α PE (clone MAb11) (BioLegend) and anti-IL-10 PE (clone JES3-9D7) (BD Biosciences).

    Techniques: Incubation, Enzyme-linked Immunosorbent Assay, In Vitro, Standard Deviation, MANN-WHITNEY

    Neutralization of IL-10 increases IFN-γ and TNF-α production by CD3 + versus CD3 − cells in response to B. pseudomallei. PBMCs of healthy seropositive individuals (n = 8) were incubated with 10 μg/ml E. coli LPS or killed B. pseudomallei (30:1 ratio) versus medium alone and in the presence or absence of 3 μg/ml of anti-IL-10 mAb. IFN-γ ( a ), TNF-α ( b ) and IL-6 ( c ) production was measured by ELISA from collected supernatant after 48 hours in vitro . Each symbol represents data from an individual. The values from the same individual in the presence or absence of anti-IL-10 mAb condition are joined by a line. To identify the cellular source of cytokines, PBMCs were incubated with killed B. pseudomallei (30:1 ratio) for 20 hours and assayed by flow cytometry for intracellular IFN-γ and TNF-α versus cell surface expression of CD3. A profile of one representative donor, gated initially on lymphocytes ( d ) and monocytes ( e ) by FSC/SSC shows the frequency of IFN-γ and TNF-α producing cells respectively. Statistical significance was determined using paired T test; ns, non significant, * p

    Journal: Scientific Reports

    Article Title: Interleukin 10 inhibits pro-inflammatory cytokine responses and killing of Burkholderia pseudomallei

    doi: 10.1038/srep42791

    Figure Lengend Snippet: Neutralization of IL-10 increases IFN-γ and TNF-α production by CD3 + versus CD3 − cells in response to B. pseudomallei. PBMCs of healthy seropositive individuals (n = 8) were incubated with 10 μg/ml E. coli LPS or killed B. pseudomallei (30:1 ratio) versus medium alone and in the presence or absence of 3 μg/ml of anti-IL-10 mAb. IFN-γ ( a ), TNF-α ( b ) and IL-6 ( c ) production was measured by ELISA from collected supernatant after 48 hours in vitro . Each symbol represents data from an individual. The values from the same individual in the presence or absence of anti-IL-10 mAb condition are joined by a line. To identify the cellular source of cytokines, PBMCs were incubated with killed B. pseudomallei (30:1 ratio) for 20 hours and assayed by flow cytometry for intracellular IFN-γ and TNF-α versus cell surface expression of CD3. A profile of one representative donor, gated initially on lymphocytes ( d ) and monocytes ( e ) by FSC/SSC shows the frequency of IFN-γ and TNF-α producing cells respectively. Statistical significance was determined using paired T test; ns, non significant, * p

    Article Snippet: Intracellular cytokine production was detected with anti-IFN-γ FITC (clone 4SB3), anti-TNF-α PE (clone MAb11) (BioLegend) and anti-IL-10 PE (clone JES3-9D7) (BD Biosciences).

    Techniques: Neutralization, Incubation, Enzyme-linked Immunosorbent Assay, In Vitro, Flow Cytometry, Cytometry, Expressing

    Neutralization of IL-10 increases IFN-γ, TNF-α and IL-6 production in individuals with DM. PBMCs (n = 5) were incubated with 10 μg/ml E. coli LPS, killed B. pseudomallei (30:1 ratio) or medium alone in the presence or absence of anti-IL-10 mAb. IFN-γ ( a ), TNF-α ( b ) and IL-6 ( c ) was measured by ELISA from collected supernatant after 48 hours in vitro . Each symbol represents data from an individual. The values from the same individual in the presence or absence of anti-IL-10 mAb are joined by a line. Statistical significance was determined using paired T test; ns, non significant, * p

    Journal: Scientific Reports

    Article Title: Interleukin 10 inhibits pro-inflammatory cytokine responses and killing of Burkholderia pseudomallei

    doi: 10.1038/srep42791

    Figure Lengend Snippet: Neutralization of IL-10 increases IFN-γ, TNF-α and IL-6 production in individuals with DM. PBMCs (n = 5) were incubated with 10 μg/ml E. coli LPS, killed B. pseudomallei (30:1 ratio) or medium alone in the presence or absence of anti-IL-10 mAb. IFN-γ ( a ), TNF-α ( b ) and IL-6 ( c ) was measured by ELISA from collected supernatant after 48 hours in vitro . Each symbol represents data from an individual. The values from the same individual in the presence or absence of anti-IL-10 mAb are joined by a line. Statistical significance was determined using paired T test; ns, non significant, * p

    Article Snippet: Intracellular cytokine production was detected with anti-IFN-γ FITC (clone 4SB3), anti-TNF-α PE (clone MAb11) (BioLegend) and anti-IL-10 PE (clone JES3-9D7) (BD Biosciences).

    Techniques: Neutralization, Incubation, Enzyme-linked Immunosorbent Assay, In Vitro

    IL-10 and IFN-γ production by whole blood of healthy individuals in response to B. pseudomallei . Whole blood of a healthy representative seropositive donor was incubated with killed B. pseudomallei (30:1 ratio) for 6, 12, 24 or 48 hours in vitro and supernatants assayed for IFN-γ and IL-10 ( a ). Whole blood of healthy seropositive donors (n = 75) was stimulated with killed B. pseudomallei (30:1 ratio) or 10 μg/ml of E. coli LPS and supernatants assayed for IFN-γ and IL-10 by ELISA after 48 hours in vitro ( b ). IFN-γ ( c ) and IL-10 ( d ) production in response to E. coli LPS or killed B. pseudomallei (30:1 ratio) from experiment performed in ( b ) were plotted according to the age of the individuals. The data in ( b ) are presented with the mean and the standard deviation. Statistical significance was determined using one way ANOVA and Tukey’s post test or Pearson correlation; ns, non significant, * p

    Journal: Scientific Reports

    Article Title: Interleukin 10 inhibits pro-inflammatory cytokine responses and killing of Burkholderia pseudomallei

    doi: 10.1038/srep42791

    Figure Lengend Snippet: IL-10 and IFN-γ production by whole blood of healthy individuals in response to B. pseudomallei . Whole blood of a healthy representative seropositive donor was incubated with killed B. pseudomallei (30:1 ratio) for 6, 12, 24 or 48 hours in vitro and supernatants assayed for IFN-γ and IL-10 ( a ). Whole blood of healthy seropositive donors (n = 75) was stimulated with killed B. pseudomallei (30:1 ratio) or 10 μg/ml of E. coli LPS and supernatants assayed for IFN-γ and IL-10 by ELISA after 48 hours in vitro ( b ). IFN-γ ( c ) and IL-10 ( d ) production in response to E. coli LPS or killed B. pseudomallei (30:1 ratio) from experiment performed in ( b ) were plotted according to the age of the individuals. The data in ( b ) are presented with the mean and the standard deviation. Statistical significance was determined using one way ANOVA and Tukey’s post test or Pearson correlation; ns, non significant, * p

    Article Snippet: Intracellular cytokine production was detected with anti-IFN-γ FITC (clone 4SB3), anti-TNF-α PE (clone MAb11) (BioLegend) and anti-IL-10 PE (clone JES3-9D7) (BD Biosciences).

    Techniques: Incubation, In Vitro, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Neutralization of IL-10 increases the killing of B. pseudomallei by healthy individuals and individuals with DM. PBMCs from healthy individuals (n = 7) ( a ) and individuals with DM (n = 8) ( b ) were incubated with medium, live B. pseudomallei at an MOI of 1 in the presence or absence of 3 ug/ml of anti-IL-10 mAb for 6 hours. The number of live B. pseudomallei was assessed by colony forming unit assay. The cultured supernatants of healthy individuals (n = 5) ( c ) and individuals with DM (n = 8) ( d ) were assayed for IFN-γ, TNF-α and IL-6 by ELISA. Each symbol represents data from an individual. The values from the same individual in the presence or absence of anti-IL-10 mAb are joined by a line. Statistical significance was determined using paired T test; ns, non significant, * p

    Journal: Scientific Reports

    Article Title: Interleukin 10 inhibits pro-inflammatory cytokine responses and killing of Burkholderia pseudomallei

    doi: 10.1038/srep42791

    Figure Lengend Snippet: Neutralization of IL-10 increases the killing of B. pseudomallei by healthy individuals and individuals with DM. PBMCs from healthy individuals (n = 7) ( a ) and individuals with DM (n = 8) ( b ) were incubated with medium, live B. pseudomallei at an MOI of 1 in the presence or absence of 3 ug/ml of anti-IL-10 mAb for 6 hours. The number of live B. pseudomallei was assessed by colony forming unit assay. The cultured supernatants of healthy individuals (n = 5) ( c ) and individuals with DM (n = 8) ( d ) were assayed for IFN-γ, TNF-α and IL-6 by ELISA. Each symbol represents data from an individual. The values from the same individual in the presence or absence of anti-IL-10 mAb are joined by a line. Statistical significance was determined using paired T test; ns, non significant, * p

    Article Snippet: Intracellular cytokine production was detected with anti-IFN-γ FITC (clone 4SB3), anti-TNF-α PE (clone MAb11) (BioLegend) and anti-IL-10 PE (clone JES3-9D7) (BD Biosciences).

    Techniques: Neutralization, Incubation, Colony-forming Unit Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

    Macrophages from myeloid-deficient MOF mice demonstrate decreased NF-κB inflammatory genes. ( A ) Representative figures showing Mof deletion in BMDMs and wound macrophages from Lyz2Cre Mof fl/fl mice. BMDMs were derived from Lyz2Cre Mof fl/fl or Mof fl/fl mice. Wound macrophages were sorted by magnetic-activated cell sorting (MACS) from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Mof gene expression was measured by qPCR ( n = 3 × 4 mice pooled/replicate, repeated twice). ( B ) Representative figures showing inflammatory cytokine gene expression in LPS-stimulated BMDMs. BMDMs were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice and treated with LPS or vehicle control for 8 hours. Il1b , Tnf , and Nos2 gene expression was measured by qPCR ( n = 3, repeated twice). ( C ) Representative figures showing baseline inflammatory cytokine gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Il1b ( n = 3), Tnf ( n = 3), and Nos2 ( n = 4) expression was measured by qPCR ( n = 3 mice pooled/replicate, repeated twice). ( D ) Representative figures showing antiinflammatory gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Il10 , Arg1 , and Ym1 expression was measured by qPCR ( n = 3 × 3 mice pooled/replicate, repeated twice). A , C , and D were analyzed using a 2-tailed Student’s t test. B was analyzed using 2-way ANOVA followed by Holm-Šídák test for multiple comparisons.

    Journal: JCI Insight

    Article Title: TNF- α regulates diabetic macrophage function through the histone acetyltransferase MOF

    doi: 10.1172/jci.insight.132306

    Figure Lengend Snippet: Macrophages from myeloid-deficient MOF mice demonstrate decreased NF-κB inflammatory genes. ( A ) Representative figures showing Mof deletion in BMDMs and wound macrophages from Lyz2Cre Mof fl/fl mice. BMDMs were derived from Lyz2Cre Mof fl/fl or Mof fl/fl mice. Wound macrophages were sorted by magnetic-activated cell sorting (MACS) from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Mof gene expression was measured by qPCR ( n = 3 × 4 mice pooled/replicate, repeated twice). ( B ) Representative figures showing inflammatory cytokine gene expression in LPS-stimulated BMDMs. BMDMs were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice and treated with LPS or vehicle control for 8 hours. Il1b , Tnf , and Nos2 gene expression was measured by qPCR ( n = 3, repeated twice). ( C ) Representative figures showing baseline inflammatory cytokine gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Il1b ( n = 3), Tnf ( n = 3), and Nos2 ( n = 4) expression was measured by qPCR ( n = 3 mice pooled/replicate, repeated twice). ( D ) Representative figures showing antiinflammatory gene expression in wound macrophages. Wound macrophages (CD11b + CD3 – CD19 – Ly6G – ) were isolated from Lyz2Cre Mof fl/fl or Mof fl/fl mice on day 5 after wounding. Il10 , Arg1 , and Ym1 expression was measured by qPCR ( n = 3 × 3 mice pooled/replicate, repeated twice). A , C , and D were analyzed using a 2-tailed Student’s t test. B was analyzed using 2-way ANOVA followed by Holm-Šídák test for multiple comparisons.

    Article Snippet: RNA was then normalized and reversed-transcribed to cDNA using Superscript III (Invitrogen, Thermo Fisher Scientific). qPCR was performed in triplicate with 2X TaqMan PCR mix using the 7500 Real-Time PCR System and primers for Il1b (Mm00434228_m10), Tnfa (Mm00443258_m1), Mof (Mm01137921_g1), Msl1 (Mm00511921_m1), Msl2 (Mm01235702_m1), Kansl1 (Mm1246891_m1), Il10 (Mm01288386_m1), Arg1 (Mm00475988_m1), Ym1 (Mm00657889_mH) (all from Thermo Fisher Scientific), and 18s as the internal control.

    Techniques: Mouse Assay, Derivative Assay, FACS, Magnetic Cell Separation, Expressing, Real-time Polymerase Chain Reaction, Isolation

    PRB782 shows improved biological activity. Induction of IL-10 production with treatment of Colo205 cells with induced bacterial supernatant from PRB484 and PRB782 showed PRB782 to have significantly (~15-fold) higher biological activity as compared to PRB484. Data represent the average of 3 biological replicates with error bars indicating standard deviation from the mean. Comparison performed with t -tests. ** P

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Challenges and Pitfalls in the Engineering of Human Interleukin 22 (hIL-22) Secreting Lactobacillus reuteri

    doi: 10.3389/fbioe.2020.00543

    Figure Lengend Snippet: PRB782 shows improved biological activity. Induction of IL-10 production with treatment of Colo205 cells with induced bacterial supernatant from PRB484 and PRB782 showed PRB782 to have significantly (~15-fold) higher biological activity as compared to PRB484. Data represent the average of 3 biological replicates with error bars indicating standard deviation from the mean. Comparison performed with t -tests. ** P

    Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA)ELISA was used to quantify the expression levels of hIL-22, human IL-10, and Reg3α.

    Techniques: Activity Assay, Standard Deviation

    Expression and secretion of hIL-22 by L. reuteri . (A) Recombinant hIL-22 detected by ELISA in supernatants of recombinant L. reuteri (PRB484) upon induction. L. reuteri expressing pSIP411 was used as control (PRB577). (B) Induction of IL-10 production from Colo205 cells upon treatment with commercial rhIL-22 as compared to bacterially secreted IL-22. Data are mean of n = 3 with error bars representing deviation from the mean; comparisons performed with t -tests (two groups) or analysis of variance (ANOVA) (multiple groups). *** P

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Challenges and Pitfalls in the Engineering of Human Interleukin 22 (hIL-22) Secreting Lactobacillus reuteri

    doi: 10.3389/fbioe.2020.00543

    Figure Lengend Snippet: Expression and secretion of hIL-22 by L. reuteri . (A) Recombinant hIL-22 detected by ELISA in supernatants of recombinant L. reuteri (PRB484) upon induction. L. reuteri expressing pSIP411 was used as control (PRB577). (B) Induction of IL-10 production from Colo205 cells upon treatment with commercial rhIL-22 as compared to bacterially secreted IL-22. Data are mean of n = 3 with error bars representing deviation from the mean; comparisons performed with t -tests (two groups) or analysis of variance (ANOVA) (multiple groups). *** P

    Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA)ELISA was used to quantify the expression levels of hIL-22, human IL-10, and Reg3α.

    Techniques: Expressing, Recombinant, Enzyme-linked Immunosorbent Assay

    Immunohistochemical expression of anti‐inflammatory cytokines (Magnification: 400×). IL‐4: A, C control placenta (35 wks, 38 wks) showing moderate cytoplasmic expression in ST; B, D, preeclamptic placenta (35 wks, 38 wks) showing mild cytoplasmic expression in ST. IL‐10: E, G, control placenta (35 wks, 38 wks) showing moderate cytoplasmic expression in ST; F, H preeclamptic placenta (32 wks, 38 wks) showing mild cytoplasmic expression in ST

    Journal: Journal of Clinical Laboratory Analysis

    Article Title: Association of pro‐ and anti‐inflammatory cytokines in preeclampsia. Association of pro‐ and anti‐inflammatory cytokines in preeclampsia

    doi: 10.1002/jcla.22834

    Figure Lengend Snippet: Immunohistochemical expression of anti‐inflammatory cytokines (Magnification: 400×). IL‐4: A, C control placenta (35 wks, 38 wks) showing moderate cytoplasmic expression in ST; B, D, preeclamptic placenta (35 wks, 38 wks) showing mild cytoplasmic expression in ST. IL‐10: E, G, control placenta (35 wks, 38 wks) showing moderate cytoplasmic expression in ST; F, H preeclamptic placenta (32 wks, 38 wks) showing mild cytoplasmic expression in ST

    Article Snippet: The sections were incubated overnight in humid chamber at 4°C with primary antibodies IL‐6 (dilution 1:200, rabbit polyclonal, Abcam Inc, Cambridge, UK), TNF‐α (dilution 1:150, rabbit polyclonal, Abcam Inc), IL‐4 (dilution 1:200, rabbit polyclonal, Abcam Inc), and IL‐10 (dilution 1:150, rabbit polyclonal, Abcam Inc).

    Techniques: Immunohistochemistry, Expressing

    Receiver operating characteristics (ROC) curve showing the expression of serum IL‐6, TNF‐α, IL‐4, IL‐10 to differentiate preeclamptic Group from control. A, B, Group I and Group II of IL‐6; C, D, Group I and Group II of TNF‐α; E, F, Group I and Group II of IL‐4; G, H, Group I and Group II of IL‐10

    Journal: Journal of Clinical Laboratory Analysis

    Article Title: Association of pro‐ and anti‐inflammatory cytokines in preeclampsia. Association of pro‐ and anti‐inflammatory cytokines in preeclampsia

    doi: 10.1002/jcla.22834

    Figure Lengend Snippet: Receiver operating characteristics (ROC) curve showing the expression of serum IL‐6, TNF‐α, IL‐4, IL‐10 to differentiate preeclamptic Group from control. A, B, Group I and Group II of IL‐6; C, D, Group I and Group II of TNF‐α; E, F, Group I and Group II of IL‐4; G, H, Group I and Group II of IL‐10

    Article Snippet: The sections were incubated overnight in humid chamber at 4°C with primary antibodies IL‐6 (dilution 1:200, rabbit polyclonal, Abcam Inc, Cambridge, UK), TNF‐α (dilution 1:150, rabbit polyclonal, Abcam Inc), IL‐4 (dilution 1:200, rabbit polyclonal, Abcam Inc), and IL‐10 (dilution 1:150, rabbit polyclonal, Abcam Inc).

    Techniques: Expressing

    Reverse transcriptase PCR analysis of the preeclamptic (P) and normal (N) placental tissues on representative samples. A, mRNA transcripts of all proteins where N1, P1 represents Group I samples and N2, N3, P2, P3 represents Group II samples. β‐actin was used as control. B, Quantification of the mRNA transcripts of IL‐6, TNF‐α, IL‐4, IL‐10 where the values in the curve represents the percentage of the samples

    Journal: Journal of Clinical Laboratory Analysis

    Article Title: Association of pro‐ and anti‐inflammatory cytokines in preeclampsia. Association of pro‐ and anti‐inflammatory cytokines in preeclampsia

    doi: 10.1002/jcla.22834

    Figure Lengend Snippet: Reverse transcriptase PCR analysis of the preeclamptic (P) and normal (N) placental tissues on representative samples. A, mRNA transcripts of all proteins where N1, P1 represents Group I samples and N2, N3, P2, P3 represents Group II samples. β‐actin was used as control. B, Quantification of the mRNA transcripts of IL‐6, TNF‐α, IL‐4, IL‐10 where the values in the curve represents the percentage of the samples

    Article Snippet: The sections were incubated overnight in humid chamber at 4°C with primary antibodies IL‐6 (dilution 1:200, rabbit polyclonal, Abcam Inc, Cambridge, UK), TNF‐α (dilution 1:150, rabbit polyclonal, Abcam Inc), IL‐4 (dilution 1:200, rabbit polyclonal, Abcam Inc), and IL‐10 (dilution 1:150, rabbit polyclonal, Abcam Inc).

    Techniques: Polymerase Chain Reaction

    LPS increases protein content of (a) IL-6 and (b) TNFα in gastrocnemius muscles of IL-10 +/+ wild-type and IL-10 −/− mice. * p

    Journal:

    Article Title: Exaggerated Expression of Skeletal Muscle-Derived Interleukin-6, but not TNF?, in Mice Lacking Interleukin-10

    doi: 10.1016/j.jneuroim.2008.05.004

    Figure Lengend Snippet: LPS increases protein content of (a) IL-6 and (b) TNFα in gastrocnemius muscles of IL-10 +/+ wild-type and IL-10 −/− mice. * p

    Article Snippet: The response to LPS was greater (p < 0.05) in the IL-10−/− mice compared to the IL-10+/+ mice at both 4 and 24 h. IL-6 mRNA expression was markedly increased relative to the corresponding saline controls for both IL-10+/+ and IL-10−/− mice at 4 h post LPS, the response of IL-10−/− mice being more that twice that of the wild-type mice.

    Techniques: Mouse Assay

    LPS increases mRNA expression of both (a) IL-6 and (b) TNFα in gastrocnemius muscles of IL-10 +/+ wild-type and IL-10 −/− mice. Data are expressed as fold changes relative to control (set at 1.0 indicated by the dashed line). * p

    Journal:

    Article Title: Exaggerated Expression of Skeletal Muscle-Derived Interleukin-6, but not TNF?, in Mice Lacking Interleukin-10

    doi: 10.1016/j.jneuroim.2008.05.004

    Figure Lengend Snippet: LPS increases mRNA expression of both (a) IL-6 and (b) TNFα in gastrocnemius muscles of IL-10 +/+ wild-type and IL-10 −/− mice. Data are expressed as fold changes relative to control (set at 1.0 indicated by the dashed line). * p

    Article Snippet: The response to LPS was greater (p < 0.05) in the IL-10−/− mice compared to the IL-10+/+ mice at both 4 and 24 h. IL-6 mRNA expression was markedly increased relative to the corresponding saline controls for both IL-10+/+ and IL-10−/− mice at 4 h post LPS, the response of IL-10−/− mice being more that twice that of the wild-type mice.

    Techniques: Expressing, Mouse Assay

    Human innate and adaptive immune cells from hNSG mice are functional and respond to cell-specific stimulation. ( A ) Human splenocytes upregulate cell surface expression of activation marker CD69 48 hours after stimulation with human CD3/28 antibody and rhIL2. ( B ) Proportion of hCD4 + and hCD8 + T cells expressing CD69 48 hours after stimulation by hCD3/28 and rhIL2. ( C ) Decrease in CFSE staining demonstrating robust proliferation of human splenocytes stimulated with hCD3/28 and rhIL2. ( D ) Proportion of proliferating splenocytes 96 hours after cell specific stimulation. ( E , F ) Quantification of human interferon gamma (IFNγ) and IL10 in culture supernatants after 96 hours of cell specific stimulation. ( G ) Human TNFα quantification in blood serum 1 hour after in vivo injection with 3 mg/kg lipopolysaccharide (LPS). Human IgG ( H ) and IgM ( I ) from blood serum in hNSG mice. Note the absence of human cytokines and antibodies in blood serum of non-engrafted NSG mice treated with LPS, demonstrating species specificity of ELISAs. ND = not detected. Data average ± SEM; n = 2 biological replicates in ( B , D ) n = 4 biological replicates in ( E , F ) n = 3 mice per group in ( G , H ) n = 3 NSG and n = 6 hNSG mice in ( I , J ).

    Journal: Scientific Reports

    Article Title: Human immune cells infiltrate the spinal cord and impair recovery after spinal cord injury in humanized mice

    doi: 10.1038/s41598-019-55729-z

    Figure Lengend Snippet: Human innate and adaptive immune cells from hNSG mice are functional and respond to cell-specific stimulation. ( A ) Human splenocytes upregulate cell surface expression of activation marker CD69 48 hours after stimulation with human CD3/28 antibody and rhIL2. ( B ) Proportion of hCD4 + and hCD8 + T cells expressing CD69 48 hours after stimulation by hCD3/28 and rhIL2. ( C ) Decrease in CFSE staining demonstrating robust proliferation of human splenocytes stimulated with hCD3/28 and rhIL2. ( D ) Proportion of proliferating splenocytes 96 hours after cell specific stimulation. ( E , F ) Quantification of human interferon gamma (IFNγ) and IL10 in culture supernatants after 96 hours of cell specific stimulation. ( G ) Human TNFα quantification in blood serum 1 hour after in vivo injection with 3 mg/kg lipopolysaccharide (LPS). Human IgG ( H ) and IgM ( I ) from blood serum in hNSG mice. Note the absence of human cytokines and antibodies in blood serum of non-engrafted NSG mice treated with LPS, demonstrating species specificity of ELISAs. ND = not detected. Data average ± SEM; n = 2 biological replicates in ( B , D ) n = 4 biological replicates in ( E , F ) n = 3 mice per group in ( G , H ) n = 3 NSG and n = 6 hNSG mice in ( I , J ).

    Article Snippet: Human IFNγ and IL10 were quantified in culture supernatants (1:2 dilution) using DuoSet ELISA kits (R & D Systems).

    Techniques: Mouse Assay, Functional Assay, Expressing, Activation Assay, Marker, Staining, In Vivo, Injection

    Secretion of IL-12/23p40 ( A ) and IFN-γ ( B ) by mesenteric lymph node (MLN) cells obtained from WT or IL-10 −/− mice fed control or curcumin-supplemented diets and stimulated with cecal antigen extract. MLN cells were stimulated by

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Secretion of IL-12/23p40 ( A ) and IFN-γ ( B ) by mesenteric lymph node (MLN) cells obtained from WT or IL-10 −/− mice fed control or curcumin-supplemented diets and stimulated with cecal antigen extract. MLN cells were stimulated by

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques: Mouse Assay

    Synergistic inhibition of IL-1β-induced NF-κB activity by IL-10 and curcumin in stably transfected Mode-K cells. Mode-K cells transfected with NF-κB-inducible reporter plasmid (pNiFty2-Luc) were determined to express detectable

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Synergistic inhibition of IL-1β-induced NF-κB activity by IL-10 and curcumin in stably transfected Mode-K cells. Mode-K cells transfected with NF-κB-inducible reporter plasmid (pNiFty2-Luc) were determined to express detectable

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques: Inhibition, Activity Assay, Stable Transfection, Transfection, Plasmid Preparation

    Secretion of IL-12/23p40 ( A ) and IFN-γ ( B ) by cultured colonic explants obtained from wild-type (WT) or IL-10 −/− mice fed control or curcumin-supplemented diets. Colonic tissues were cultured for 18 h, and concentration of the

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Secretion of IL-12/23p40 ( A ) and IFN-γ ( B ) by cultured colonic explants obtained from wild-type (WT) or IL-10 −/− mice fed control or curcumin-supplemented diets. Colonic tissues were cultured for 18 h, and concentration of the

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques: Cell Culture, Mouse Assay, Concentration Assay

    Synergistic inhibition of LPS-induced IL-12/23p40 production by IL-10 and submicromolar concentrations of curcumin in dendritic cells in vitro. Dose response to increasing concentrations of curcumin in DC2.4 cells stimulated with LPS (100 ng/ml) for 24

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Synergistic inhibition of LPS-induced IL-12/23p40 production by IL-10 and submicromolar concentrations of curcumin in dendritic cells in vitro. Dose response to increasing concentrations of curcumin in DC2.4 cells stimulated with LPS (100 ng/ml) for 24

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques: Inhibition, In Vitro

    Histological scoring of the proximal ( A ) and distal colon ( B ). Values represent the mean of 5 WT animals and 7 IL-10 −/− animals

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Histological scoring of the proximal ( A ) and distal colon ( B ). Values represent the mean of 5 WT animals and 7 IL-10 −/− animals

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques:

    Scoring summary of phospho-Ser 276 p65-positive cells in the proximal ( A ) and distal colon ( B ) of WT or IL-10 −/− mice fed control or curcumin-supplemented diets. Staining was scored by counting the number of phospho-Ser 276 p65-positive cells;

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Scoring summary of phospho-Ser 276 p65-positive cells in the proximal ( A ) and distal colon ( B ) of WT or IL-10 −/− mice fed control or curcumin-supplemented diets. Staining was scored by counting the number of phospho-Ser 276 p65-positive cells;

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques: Mouse Assay, Staining

    Evaluation of enhanced green fluorescent protein (EGFP) fluorescence in the colon of IL-10 wt/wt /NF-κB EGFP and IL-10 −/− /NF-κB EGFP mice fed control or 1% curcumin-supplemented diet. Germ-free mice were colonized with

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Evaluation of enhanced green fluorescent protein (EGFP) fluorescence in the colon of IL-10 wt/wt /NF-κB EGFP and IL-10 −/− /NF-κB EGFP mice fed control or 1% curcumin-supplemented diet. Germ-free mice were colonized with

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques: Fluorescence, Mouse Assay

    Inhibition of IL-12/23p40 secretion in LPS-stimulated splenocytes from WT or IL-10 −/− mice by curcumin. Splenocytes from WT and IL-10 −/− mice were treated with curcumin (0.5 to 10 μM) for 2 h prior to LPS stimulation

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Inhibition of IL-12/23p40 secretion in LPS-stimulated splenocytes from WT or IL-10 −/− mice by curcumin. Splenocytes from WT and IL-10 −/− mice were treated with curcumin (0.5 to 10 μM) for 2 h prior to LPS stimulation

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques: Inhibition, Mouse Assay

    Histology of the proximal and distal colon or wild type (WT) and IL-10 −/− mice fed control or curcumin-supplemented diets. A : hematoxylin and eosin (H E) staining of proximal and distal colon from WT mice fed control diet. B : H E

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Histology of the proximal and distal colon or wild type (WT) and IL-10 −/− mice fed control or curcumin-supplemented diets. A : hematoxylin and eosin (H E) staining of proximal and distal colon from WT mice fed control diet. B : H E

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques: Mouse Assay, Staining

    Immunohistochemical analysis of the effects of dietary curcumin on NF-κB activation (phospho-Ser 276 p65) in the proximal and distal colon of IL-10 −/− mice in situ. Colonic tissues from wild-type mice fed control diet ( A ) and IL-10

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Limited effects of dietary curcumin on Th-1 driven colitis in IL-10 deficient mice suggest an IL-10-dependent mechanism of protection

    doi: 10.1152/ajpgi.90365.2008

    Figure Lengend Snippet: Immunohistochemical analysis of the effects of dietary curcumin on NF-κB activation (phospho-Ser 276 p65) in the proximal and distal colon of IL-10 −/− mice in situ. Colonic tissues from wild-type mice fed control diet ( A ) and IL-10

    Article Snippet: Specific pathogen-free (SPF) wild-type (WT) 129/SvEv mice and germ-free IL-10−/− mice on the same genetic background were obtained from the National Gnotobiotic Rodent Resource Center at the University of North Carolina, Chapel Hill.

    Techniques: Immunohistochemistry, Activation Assay, Mouse Assay, In Situ

    LPS-induced ROS production is p47 phox dependent in macrophages and ROS scavenger blocks IL-10 secretion from HLL mice. A , Representative tracing showed the generation of superoxide as counts per second (CPS) by peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice, stimulated with 100 ng/ml of LPS. B , Peritoneal macrophages from HLL mice were incubated with DPI for 30 min and the cells were left unstimulated or stimulated with LPS (100 ng/ml) for an additional 8 h. IL-10 secretion in the culture supernatant was detected by ELISA. C, E , and F , Peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice were treated with LPS (100 ng/ml) for 8 h. IL-10, IL-6, and TNF-α were measured in the culture supernatant. Results represent mean ± SEM of three separate experiments and *, p

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

    Article Title: Protective Role of Reactive Oxygen Species in Endotoxin-Induced Lung Inflammation through Modulation of IL-10 Expression

    doi: 10.4049/jimmunol.1101323

    Figure Lengend Snippet: LPS-induced ROS production is p47 phox dependent in macrophages and ROS scavenger blocks IL-10 secretion from HLL mice. A , Representative tracing showed the generation of superoxide as counts per second (CPS) by peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice, stimulated with 100 ng/ml of LPS. B , Peritoneal macrophages from HLL mice were incubated with DPI for 30 min and the cells were left unstimulated or stimulated with LPS (100 ng/ml) for an additional 8 h. IL-10 secretion in the culture supernatant was detected by ELISA. C, E , and F , Peritoneal macrophages from p47 phox−/− /HLL mice and HLL mice were treated with LPS (100 ng/ml) for 8 h. IL-10, IL-6, and TNF-α were measured in the culture supernatant. Results represent mean ± SEM of three separate experiments and *, p

    Article Snippet: In another group, each mouse was administered (i.p.) 1 μg of recombinant IL-10 (R & D Systems Inc., MN, USA) 2 h prior to LPS treatment.

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

    Macrophages from p47 phox deficient mice have decreased phosphorylation of Akt and GSK3-β induced by LPS, which modulates IL-10 production by inhibition of Akt and GSK3. Bone Marrow Macrophages were derived from p47 phox−/− /HLL or HLL mice for 6 d. A , The cells were serum-starved for 4 h, then stimulated with 100 ng/ml LPS for 30 and 60 min or left unstimulated. B , Macrophages from HLL mice were starved for 4 h and then preincubated with 5 μM Akt inhibitor (Akt-i) for 1 h before stimulation with 100 ng/ml LPS or left unstimulated. Akt and GSK3- β phosphorylation were assessed using equal amounts of protein by Western blotting with antibodies against Akt-total and GSK3-β-total. Data are representative of 3 separate experiments. C, D , IL-10 production in macrophages isolated from HLL mice ( C ) and p47 phox−/− /HLL mice ( D ) were preincubated for 1 h with PBS, DMSO, 20 μM Akt inhibitor or 10 μM SB216763 and then left unstimulated or stimulated with 100 ng/ml LPS for 8 h. Cell-free supernatants were analyzed by ELISA. Results represent mean ± SEM and *, p

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

    Article Title: Protective Role of Reactive Oxygen Species in Endotoxin-Induced Lung Inflammation through Modulation of IL-10 Expression

    doi: 10.4049/jimmunol.1101323

    Figure Lengend Snippet: Macrophages from p47 phox deficient mice have decreased phosphorylation of Akt and GSK3-β induced by LPS, which modulates IL-10 production by inhibition of Akt and GSK3. Bone Marrow Macrophages were derived from p47 phox−/− /HLL or HLL mice for 6 d. A , The cells were serum-starved for 4 h, then stimulated with 100 ng/ml LPS for 30 and 60 min or left unstimulated. B , Macrophages from HLL mice were starved for 4 h and then preincubated with 5 μM Akt inhibitor (Akt-i) for 1 h before stimulation with 100 ng/ml LPS or left unstimulated. Akt and GSK3- β phosphorylation were assessed using equal amounts of protein by Western blotting with antibodies against Akt-total and GSK3-β-total. Data are representative of 3 separate experiments. C, D , IL-10 production in macrophages isolated from HLL mice ( C ) and p47 phox−/− /HLL mice ( D ) were preincubated for 1 h with PBS, DMSO, 20 μM Akt inhibitor or 10 μM SB216763 and then left unstimulated or stimulated with 100 ng/ml LPS for 8 h. Cell-free supernatants were analyzed by ELISA. Results represent mean ± SEM and *, p

    Article Snippet: In another group, each mouse was administered (i.p.) 1 μg of recombinant IL-10 (R & D Systems Inc., MN, USA) 2 h prior to LPS treatment.

    Techniques: Mouse Assay, Inhibition, Derivative Assay, Western Blot, Isolation, Enzyme-linked Immunosorbent Assay

    Recombinant IL-10 can substantially reduce the lung inflammation induced by LPS in p47 phox−/− /HLL mice. A , Bioluminescent detection of NF-kB activation in p47 phox−/− /HLL mice was performed. After a single i.p. injection of recombinant IL-10 (1 μg/mouse), LPS were administered by i.p. injection at 2 h and then bioluminescence was measured at 0, 8, and 24 h following luciferin injection. Images are representative of 3 mice per group. B , Photons were emitted over the chest in previous groups at 0, 8, and 24 h. Results represent mean ± SEM and **, p

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

    Article Title: Protective Role of Reactive Oxygen Species in Endotoxin-Induced Lung Inflammation through Modulation of IL-10 Expression

    doi: 10.4049/jimmunol.1101323

    Figure Lengend Snippet: Recombinant IL-10 can substantially reduce the lung inflammation induced by LPS in p47 phox−/− /HLL mice. A , Bioluminescent detection of NF-kB activation in p47 phox−/− /HLL mice was performed. After a single i.p. injection of recombinant IL-10 (1 μg/mouse), LPS were administered by i.p. injection at 2 h and then bioluminescence was measured at 0, 8, and 24 h following luciferin injection. Images are representative of 3 mice per group. B , Photons were emitted over the chest in previous groups at 0, 8, and 24 h. Results represent mean ± SEM and **, p

    Article Snippet: In another group, each mouse was administered (i.p.) 1 μg of recombinant IL-10 (R & D Systems Inc., MN, USA) 2 h prior to LPS treatment.

    Techniques: Recombinant, Mouse Assay, Activation Assay, Injection

    Schematic of adaptive immunoregulation of LUT and chlorogenic acid in LPS-induced IL-10 expression. CGA enhanced LPS-induced expression of IL-10 and IL-6 and increased the expression of NF-κB, Sp1, C/EBPβ, and δ. The effect of CGA was interfered with LUT by suppressing the phosphorylation of IκB and p38 and downregulated NF-κB activity. In the event, IL-6 was suppressed and IL-10 was not influenced. LPS: Lipopolysaccharide, CGA: Chlorogenic acid, LUT: Luteolin, NF-κB: Nuclear factor-κB, IL-10: Interleukin-10

    Journal: Tzu-Chi Medical Journal

    Article Title: Adaptive immunoregulation of luteolin and chlorogenic acid in lipopolysaccharide-induced interleukin-10 expression

    doi: 10.4103/tcmj.tcmj_23_19

    Figure Lengend Snippet: Schematic of adaptive immunoregulation of LUT and chlorogenic acid in LPS-induced IL-10 expression. CGA enhanced LPS-induced expression of IL-10 and IL-6 and increased the expression of NF-κB, Sp1, C/EBPβ, and δ. The effect of CGA was interfered with LUT by suppressing the phosphorylation of IκB and p38 and downregulated NF-κB activity. In the event, IL-6 was suppressed and IL-10 was not influenced. LPS: Lipopolysaccharide, CGA: Chlorogenic acid, LUT: Luteolin, NF-κB: Nuclear factor-κB, IL-10: Interleukin-10

    Article Snippet: SP-1, C/EBPβ, C/EBPd, and enzyme-linked immunosorbent assay (ELISA) kits of IL-10 and IL-6 were obtained from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA.

    Techniques: Expressing, Activity Assay

    GGS and LJF affect LPS-induced inflammatory factor in RAW264.7 cells. RAW cells were challenged at baseline with LPS (10 ng/mL) or none ( n = 6). Raw cells receiving LPS were pretreated with 100 μg/mL GGS ( n = 6) and 10 μg/mL LJF. After 24 h, the cells were obtained and assayed. The mRNA of cells was collected, and pro-inflammatory cytokines and IL-10 were evaluated by RT-PCR. RT-PCR: Reverse transcription-polymerase chain reaction, LPS: Lipopolysaccharide, IL-10: Interleukin-10, GGS: Gingyo-san, LJF: Lonicerae japonica flos

    Journal: Tzu-Chi Medical Journal

    Article Title: Adaptive immunoregulation of luteolin and chlorogenic acid in lipopolysaccharide-induced interleukin-10 expression

    doi: 10.4103/tcmj.tcmj_23_19

    Figure Lengend Snippet: GGS and LJF affect LPS-induced inflammatory factor in RAW264.7 cells. RAW cells were challenged at baseline with LPS (10 ng/mL) or none ( n = 6). Raw cells receiving LPS were pretreated with 100 μg/mL GGS ( n = 6) and 10 μg/mL LJF. After 24 h, the cells were obtained and assayed. The mRNA of cells was collected, and pro-inflammatory cytokines and IL-10 were evaluated by RT-PCR. RT-PCR: Reverse transcription-polymerase chain reaction, LPS: Lipopolysaccharide, IL-10: Interleukin-10, GGS: Gingyo-san, LJF: Lonicerae japonica flos

    Article Snippet: SP-1, C/EBPβ, C/EBPd, and enzyme-linked immunosorbent assay (ELISA) kits of IL-10 and IL-6 were obtained from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA.

    Techniques: Reverse Transcription Polymerase Chain Reaction

    CGA and LUT affect LPS-induced IL-10 (a, b) and IL-6 (c, d) expression. Concentrations of IL-10 and IL-6 in the supernatant were measured in the culture medium obtained 24 h after LPS treatment (10 ng/mL) or none ( n = 6). The cells receiving LPS were pretreated with different doses of CGA and LUT ( n = 6). * P

    Journal: Tzu-Chi Medical Journal

    Article Title: Adaptive immunoregulation of luteolin and chlorogenic acid in lipopolysaccharide-induced interleukin-10 expression

    doi: 10.4103/tcmj.tcmj_23_19

    Figure Lengend Snippet: CGA and LUT affect LPS-induced IL-10 (a, b) and IL-6 (c, d) expression. Concentrations of IL-10 and IL-6 in the supernatant were measured in the culture medium obtained 24 h after LPS treatment (10 ng/mL) or none ( n = 6). The cells receiving LPS were pretreated with different doses of CGA and LUT ( n = 6). * P

    Article Snippet: SP-1, C/EBPβ, C/EBPd, and enzyme-linked immunosorbent assay (ELISA) kits of IL-10 and IL-6 were obtained from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA.

    Techniques: Expressing

    The synchronizing action of CGA and LUT on LPS-induced IL-10 (a) and IL-6 (b) expression. Concentrations of IL-10 and IL-6 in the supernatant were measured in the culture medium obtained 24 h after LPS treatment (10 ng/ml) or none ( n = 6). The cells receiving LPS were pretreated with 10 μM CGA and/or 1 μM LUT ( n = 6). Mean ± SEM. * P

    Journal: Tzu-Chi Medical Journal

    Article Title: Adaptive immunoregulation of luteolin and chlorogenic acid in lipopolysaccharide-induced interleukin-10 expression

    doi: 10.4103/tcmj.tcmj_23_19

    Figure Lengend Snippet: The synchronizing action of CGA and LUT on LPS-induced IL-10 (a) and IL-6 (b) expression. Concentrations of IL-10 and IL-6 in the supernatant were measured in the culture medium obtained 24 h after LPS treatment (10 ng/ml) or none ( n = 6). The cells receiving LPS were pretreated with 10 μM CGA and/or 1 μM LUT ( n = 6). Mean ± SEM. * P

    Article Snippet: SP-1, C/EBPβ, C/EBPd, and enzyme-linked immunosorbent assay (ELISA) kits of IL-10 and IL-6 were obtained from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA.

    Techniques: Expressing

    The herbal effect of LPS-induced IL-10 expression on RAW264.7 cells. IL-10 was induced by LPS (10 ng/ml) at 24 h and pretreated with GGS for 1 h (▪) and ten herbs on RAW264.7 cell by ELISA kit. The ten herbs include Lonicera japonica . (♦), Forsythia suspense (▴), Mentha haplocalyx (○), Schizonepeta tenuifolia (•), Glycine max (◊), Glycyrrhiza uralensis (△), Platycodon grandiflorum (□), Lophatherum gracile (▾), Arctium lappa L. (+), and Phragmites communis (▽). All data were means of triplicates, and numbers in parentheses indicate the standard deviation of triplicates ( n = 6). LPS: Lipopolysaccharide, IL-10: Interleukin-10, GGS: Gingyo-san

    Journal: Tzu-Chi Medical Journal

    Article Title: Adaptive immunoregulation of luteolin and chlorogenic acid in lipopolysaccharide-induced interleukin-10 expression

    doi: 10.4103/tcmj.tcmj_23_19

    Figure Lengend Snippet: The herbal effect of LPS-induced IL-10 expression on RAW264.7 cells. IL-10 was induced by LPS (10 ng/ml) at 24 h and pretreated with GGS for 1 h (▪) and ten herbs on RAW264.7 cell by ELISA kit. The ten herbs include Lonicera japonica . (♦), Forsythia suspense (▴), Mentha haplocalyx (○), Schizonepeta tenuifolia (•), Glycine max (◊), Glycyrrhiza uralensis (△), Platycodon grandiflorum (□), Lophatherum gracile (▾), Arctium lappa L. (+), and Phragmites communis (▽). All data were means of triplicates, and numbers in parentheses indicate the standard deviation of triplicates ( n = 6). LPS: Lipopolysaccharide, IL-10: Interleukin-10, GGS: Gingyo-san

    Article Snippet: SP-1, C/EBPβ, C/EBPd, and enzyme-linked immunosorbent assay (ELISA) kits of IL-10 and IL-6 were obtained from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA.

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Serum levels of IL-10 and IL-17 in ulcerative colitis patients and healthy subjects were detected with ELISA. IL-10 and IL-17 levels were significantly higher, compared with control group, ∗ P

    Journal: Canadian Journal of Gastroenterology & Hepatology

    Article Title: Decreased Breg/Th17 Ratio Improved the Prognosis of Patients with Ulcerative Colitis

    doi: 10.1155/2018/5760849

    Figure Lengend Snippet: Serum levels of IL-10 and IL-17 in ulcerative colitis patients and healthy subjects were detected with ELISA. IL-10 and IL-17 levels were significantly higher, compared with control group, ∗ P

    Article Snippet: After three washes with PBS, samples were blocked with goat serum for 30 minutes and incubated with anti-IL-10 (Abcam, ab134742) and anti-IL-17 (Abcam, ab92486) at 4°C overnight.

    Techniques: Enzyme-linked Immunosorbent Assay

    B10 cell and Th17 cell counts in ulcerative colitis patients and healthy subjects. (a-b) Gating criteria to define the CD24 + CD27 + CD38 + IL-10 + Breg cell population. Different cell subsets were distinguished according to different cell labels. (c-d) The percentage of CD4 + IL-17 + cells in peripheral blood. Compared with control group, ∗ P

    Journal: Canadian Journal of Gastroenterology & Hepatology

    Article Title: Decreased Breg/Th17 Ratio Improved the Prognosis of Patients with Ulcerative Colitis

    doi: 10.1155/2018/5760849

    Figure Lengend Snippet: B10 cell and Th17 cell counts in ulcerative colitis patients and healthy subjects. (a-b) Gating criteria to define the CD24 + CD27 + CD38 + IL-10 + Breg cell population. Different cell subsets were distinguished according to different cell labels. (c-d) The percentage of CD4 + IL-17 + cells in peripheral blood. Compared with control group, ∗ P

    Article Snippet: After three washes with PBS, samples were blocked with goat serum for 30 minutes and incubated with anti-IL-10 (Abcam, ab134742) and anti-IL-17 (Abcam, ab92486) at 4°C overnight.

    Techniques:

    The expression of IL-10 and ROR γ T in ulcerative colitis patients and healthy subjects was upregulated. (a) The mRNA expression levels of IL-10 and ROR γ T were determined with qRT-PCR. (b) IL-10 and IL-17 expression by immunohistochemistry. Compared with control group, ∗ P

    Journal: Canadian Journal of Gastroenterology & Hepatology

    Article Title: Decreased Breg/Th17 Ratio Improved the Prognosis of Patients with Ulcerative Colitis

    doi: 10.1155/2018/5760849

    Figure Lengend Snippet: The expression of IL-10 and ROR γ T in ulcerative colitis patients and healthy subjects was upregulated. (a) The mRNA expression levels of IL-10 and ROR γ T were determined with qRT-PCR. (b) IL-10 and IL-17 expression by immunohistochemistry. Compared with control group, ∗ P

    Article Snippet: After three washes with PBS, samples were blocked with goat serum for 30 minutes and incubated with anti-IL-10 (Abcam, ab134742) and anti-IL-17 (Abcam, ab92486) at 4°C overnight.

    Techniques: Expressing, Quantitative RT-PCR, Immunohistochemistry

    Correlation of cell counts and cytokine levels with Mayo scoring in ulcerative colitis patients, B10/Th17 ratio, and Mayo scoring were positively correlated; serum IL-10/IL-17 ratio and Mayo scoring were positively correlated. (a) Peripheral blood B10/Th17 ratio was positively correlated with Mayo in ulcerative colitis patients ( r = 0.758, P

    Journal: Canadian Journal of Gastroenterology & Hepatology

    Article Title: Decreased Breg/Th17 Ratio Improved the Prognosis of Patients with Ulcerative Colitis

    doi: 10.1155/2018/5760849

    Figure Lengend Snippet: Correlation of cell counts and cytokine levels with Mayo scoring in ulcerative colitis patients, B10/Th17 ratio, and Mayo scoring were positively correlated; serum IL-10/IL-17 ratio and Mayo scoring were positively correlated. (a) Peripheral blood B10/Th17 ratio was positively correlated with Mayo in ulcerative colitis patients ( r = 0.758, P

    Article Snippet: After three washes with PBS, samples were blocked with goat serum for 30 minutes and incubated with anti-IL-10 (Abcam, ab134742) and anti-IL-17 (Abcam, ab92486) at 4°C overnight.

    Techniques:

    The differences in the cell counts and related cytokine levels between ulcerative colitis patients in remission and nonremission after treatment were analyzed with flow cytometry and ELISA. (a) Gating criteria to define the CD24 + CD27 + CD38 + IL-10 + Breg cell population in the remission group and nonremission group. (b) The percentage of Th17 cells in the remission group and nonremission. (c) B10/Th17 ratio in the remission group and nonremission group. (d) Serum IL-10 levels and IL-17 levels were detected with ELISA. (e) The IL-10/IL-17 ratio in the remission group and nonremission group. (f) IL-10 and IL-17 expression by immunohistochemistry in in the remission group and nonremission group. Compared with remission group, ∗ P

    Journal: Canadian Journal of Gastroenterology & Hepatology

    Article Title: Decreased Breg/Th17 Ratio Improved the Prognosis of Patients with Ulcerative Colitis

    doi: 10.1155/2018/5760849

    Figure Lengend Snippet: The differences in the cell counts and related cytokine levels between ulcerative colitis patients in remission and nonremission after treatment were analyzed with flow cytometry and ELISA. (a) Gating criteria to define the CD24 + CD27 + CD38 + IL-10 + Breg cell population in the remission group and nonremission group. (b) The percentage of Th17 cells in the remission group and nonremission. (c) B10/Th17 ratio in the remission group and nonremission group. (d) Serum IL-10 levels and IL-17 levels were detected with ELISA. (e) The IL-10/IL-17 ratio in the remission group and nonremission group. (f) IL-10 and IL-17 expression by immunohistochemistry in in the remission group and nonremission group. Compared with remission group, ∗ P

    Article Snippet: After three washes with PBS, samples were blocked with goat serum for 30 minutes and incubated with anti-IL-10 (Abcam, ab134742) and anti-IL-17 (Abcam, ab92486) at 4°C overnight.

    Techniques: Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemistry

    The effect of amino acid substitution at the RRCHR region of mouse IL-10 on biological activities. (A) The secondary structure of IL-10 mutants by CD spectroscopy adopted from unpublished data ( 36 ). (B) The ED50 of natural IL-10 was calculated as flow: Nm RRCHR (un-mutant) (black line) (0.1 ng/mL), Nm RACHR (green line) (0.28 ng/mL), Nm AACHR (blue line) (0.96 ng/mL), and Nm ARCHA (red line) (59.28 ng/mL). (C) The activation of cellular signaling (p-STAT3) determines by western blot after lysate from spleen cells of the C57BL/6 mouse treated Nm mutants and un-mutant (control) in dose-response and using the total STAT3 as an internal control. (D) The ED50 of stable IL-10 was calculated as flow: STm RRCHR (un-mutant) (black line) (0.36 ng/mL), STm RACHR (green line) (1.6 ng/mL), STm AACHR (blue line) (2.55 ng/mL), and ST ARCHA (red line) (3.02 ng/mL). (E) The activation p-STAT3 determines by western blot from spleen cells lysate of the C57BL/6 mouse treated STm mutants and un-mutant (control) in dose-response and using the total STAT3 as an internal control. The data represented as AUC (Area Under the Curve). All data are representative of two independent experiments, with triplicate cultures per experiment ( N = 2, n = 3), and bars represent standard error of the mean.

    Journal: Frontiers in Immunology

    Article Title: The Generation of an Engineered Interleukin-10 Protein With Improved Stability and Biological Function

    doi: 10.3389/fimmu.2020.01794

    Figure Lengend Snippet: The effect of amino acid substitution at the RRCHR region of mouse IL-10 on biological activities. (A) The secondary structure of IL-10 mutants by CD spectroscopy adopted from unpublished data ( 36 ). (B) The ED50 of natural IL-10 was calculated as flow: Nm RRCHR (un-mutant) (black line) (0.1 ng/mL), Nm RACHR (green line) (0.28 ng/mL), Nm AACHR (blue line) (0.96 ng/mL), and Nm ARCHA (red line) (59.28 ng/mL). (C) The activation of cellular signaling (p-STAT3) determines by western blot after lysate from spleen cells of the C57BL/6 mouse treated Nm mutants and un-mutant (control) in dose-response and using the total STAT3 as an internal control. (D) The ED50 of stable IL-10 was calculated as flow: STm RRCHR (un-mutant) (black line) (0.36 ng/mL), STm RACHR (green line) (1.6 ng/mL), STm AACHR (blue line) (2.55 ng/mL), and ST ARCHA (red line) (3.02 ng/mL). (E) The activation p-STAT3 determines by western blot from spleen cells lysate of the C57BL/6 mouse treated STm mutants and un-mutant (control) in dose-response and using the total STAT3 as an internal control. The data represented as AUC (Area Under the Curve). All data are representative of two independent experiments, with triplicate cultures per experiment ( N = 2, n = 3), and bars represent standard error of the mean.

    Article Snippet: The nine amino acids (-G3 SG4 S-) and 13 amino acid linker(-G3 SG4 -SG4-) were generated by inserting the synthetic insert into the second monomer of mouse IL-10.

    Techniques: Spectroscopy, Mutagenesis, Activation Assay, Western Blot

    Generation and characterization of the stable version of human IL-10. (A) The effect of STh (with different linkers length) and Nh on LPS-induce luciferase activity on BMDMs of transgenic mouse: the ED 50 of Nh(black line) is 0.87 ng/mL and the ED 50 of STm proteins: STh7 (green) STh9 (blue), and STh13 (red) is calculated as 0.82, 1, and 1.2 ng/mL, respectively. (B) Splenocyte lysates of the C57BL/6 mouse either unstimulated (∅) or stimulated with IL-10 (Natural type or Stable) in a dose-dependent manner. (C) The effect mouse antiIL-10 receptor antibody (clone 1B1.3a) on blocking the suppression induced by Nh and STh (10 ng/mL) in a dose-dependent fashion. Furthermore, a significant difference is also calculated to compare Nm with STm after antiIL-10Ra treatment; ANOVA. (D) Luciferase activity represented as AUC from LPS-induced BMDMs co-treated with either Nh (black line) and STh (blue line) at 10 ng/mL after incubated at 37°C in the time course. (E) Both Nh and STh were incubated at 55°C in time course, and the luciferase activity was calculated after LPS-stimulated and co-treated with either Nm (black line) and STm (blue line) at 10 ng/mL on BMDMs. (F,G) LPS-induced Luciferase activity when co-stimulated with either heat-treated at 55°C for 10 min (10 min) or untreated (0 min) of Nh and STh. The ED 50 is calculated as follow: (F) Nh heat-treated for (3.74 ng/mL) or untreated (0.17 ng/mL); (G) STh heat-treated for (0.10 ng/mL) or untreated (0.12 ng/mL). (H) Both Nh and STh at 100 ng/mL were pre-incubated with different pH buffers at 4°C for 24 h flowed by buffer exchange columns. BMDMs of the transgenic mouse then stimulated with LPS (10 ng/mL) and pH-treated IL-10 (Nh and STh) at 10 ng/mL. The percentage of luciferase activities is relative to the maximum LPS induction of luciferase. The significant difference compared between pH treatments with neutral pH (pH7) of Nh and STh on LPS-stimulated cells; *** p

    Journal: Frontiers in Immunology

    Article Title: The Generation of an Engineered Interleukin-10 Protein With Improved Stability and Biological Function

    doi: 10.3389/fimmu.2020.01794

    Figure Lengend Snippet: Generation and characterization of the stable version of human IL-10. (A) The effect of STh (with different linkers length) and Nh on LPS-induce luciferase activity on BMDMs of transgenic mouse: the ED 50 of Nh(black line) is 0.87 ng/mL and the ED 50 of STm proteins: STh7 (green) STh9 (blue), and STh13 (red) is calculated as 0.82, 1, and 1.2 ng/mL, respectively. (B) Splenocyte lysates of the C57BL/6 mouse either unstimulated (∅) or stimulated with IL-10 (Natural type or Stable) in a dose-dependent manner. (C) The effect mouse antiIL-10 receptor antibody (clone 1B1.3a) on blocking the suppression induced by Nh and STh (10 ng/mL) in a dose-dependent fashion. Furthermore, a significant difference is also calculated to compare Nm with STm after antiIL-10Ra treatment; ANOVA. (D) Luciferase activity represented as AUC from LPS-induced BMDMs co-treated with either Nh (black line) and STh (blue line) at 10 ng/mL after incubated at 37°C in the time course. (E) Both Nh and STh were incubated at 55°C in time course, and the luciferase activity was calculated after LPS-stimulated and co-treated with either Nm (black line) and STm (blue line) at 10 ng/mL on BMDMs. (F,G) LPS-induced Luciferase activity when co-stimulated with either heat-treated at 55°C for 10 min (10 min) or untreated (0 min) of Nh and STh. The ED 50 is calculated as follow: (F) Nh heat-treated for (3.74 ng/mL) or untreated (0.17 ng/mL); (G) STh heat-treated for (0.10 ng/mL) or untreated (0.12 ng/mL). (H) Both Nh and STh at 100 ng/mL were pre-incubated with different pH buffers at 4°C for 24 h flowed by buffer exchange columns. BMDMs of the transgenic mouse then stimulated with LPS (10 ng/mL) and pH-treated IL-10 (Nh and STh) at 10 ng/mL. The percentage of luciferase activities is relative to the maximum LPS induction of luciferase. The significant difference compared between pH treatments with neutral pH (pH7) of Nh and STh on LPS-stimulated cells; *** p

    Article Snippet: The nine amino acids (-G3 SG4 S-) and 13 amino acid linker(-G3 SG4 -SG4-) were generated by inserting the synthetic insert into the second monomer of mouse IL-10.

    Techniques: Luciferase, Activity Assay, Transgenic Assay, Blocking Assay, Incubation, Buffer Exchange

    Detection of the in vivo activity of the STm by suppression of LPS-induced cytokine release. (A,B) 100 μl PBS containing 25 μg LPS i.v. Injected in C57BL/6 mice (8–12 weeks). At the indicated time points, blood was taken retro-orbitally, and ELISA determined the TNF-a serum concentration, *n.d.: not detectable. (A) To establish, LPS, PBS, Nm, or STm (2 μg protein each) were injected one at a time. (B) 30 min before LPS administration, the indicated amounts of Nm or STm were i.v. Injected. (C) IL-10 -/- mice were given 10 μg LPS together with increasing concentrations of Nm or STm i.v. Injected. After 3 h, blood was drawn, and the IL 6 serum concentration determined by ELISA.

    Journal: Frontiers in Immunology

    Article Title: The Generation of an Engineered Interleukin-10 Protein With Improved Stability and Biological Function

    doi: 10.3389/fimmu.2020.01794

    Figure Lengend Snippet: Detection of the in vivo activity of the STm by suppression of LPS-induced cytokine release. (A,B) 100 μl PBS containing 25 μg LPS i.v. Injected in C57BL/6 mice (8–12 weeks). At the indicated time points, blood was taken retro-orbitally, and ELISA determined the TNF-a serum concentration, *n.d.: not detectable. (A) To establish, LPS, PBS, Nm, or STm (2 μg protein each) were injected one at a time. (B) 30 min before LPS administration, the indicated amounts of Nm or STm were i.v. Injected. (C) IL-10 -/- mice were given 10 μg LPS together with increasing concentrations of Nm or STm i.v. Injected. After 3 h, blood was drawn, and the IL 6 serum concentration determined by ELISA.

    Article Snippet: The nine amino acids (-G3 SG4 S-) and 13 amino acid linker(-G3 SG4 -SG4-) were generated by inserting the synthetic insert into the second monomer of mouse IL-10.

    Techniques: In Vivo, Activity Assay, Injection, Mouse Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

    Scheme representing the proposed biological activity and stability of stable IL-10 dimer. Both natural and stable IL-10 are biologically active in vitro and in vivo . We predict that our stable IL-10 is folded like the natural IL-10 in a domain-swapping fashion. Our mutant IL-10 model demonstrates the importance of dimerization to elicit the maximum inhibitory response. We showed that stable IL-10 is more active after treatment with different physiological stress conditions such as high-temperature and pH compared to natural IL-10.

    Journal: Frontiers in Immunology

    Article Title: The Generation of an Engineered Interleukin-10 Protein With Improved Stability and Biological Function

    doi: 10.3389/fimmu.2020.01794

    Figure Lengend Snippet: Scheme representing the proposed biological activity and stability of stable IL-10 dimer. Both natural and stable IL-10 are biologically active in vitro and in vivo . We predict that our stable IL-10 is folded like the natural IL-10 in a domain-swapping fashion. Our mutant IL-10 model demonstrates the importance of dimerization to elicit the maximum inhibitory response. We showed that stable IL-10 is more active after treatment with different physiological stress conditions such as high-temperature and pH compared to natural IL-10.

    Article Snippet: The nine amino acids (-G3 SG4 S-) and 13 amino acid linker(-G3 SG4 -SG4-) were generated by inserting the synthetic insert into the second monomer of mouse IL-10.

    Techniques: Activity Assay, In Vitro, In Vivo, Mutagenesis

    Stable mouse IL-10 (STm) is biologically active in vitro . (A) Specific induction of STAT3 phosphorylation by STm (5 ng/mL) was verified by the use of spleen cells from different mouse lines: Wild type C57BL/6 (WT), IL-10 knockout (IL-10 −/− ) and IL-10 receptor knockout (IL-10R1 −/− ). (B) Luciferase activity was monitored as the area under the curve (AUC) from BMDMs reporter mouse; cells were either unstimulated or stimulated with LPS (10 ng/mL) in the presence or absence of IL-10 from HEK 293 EBNA supernatants. A commercial mouse IL-10 (CmIL-10) was used as a positive control, and un-transfected cells supernatants (Medium) served as a negative control. Significant difference considered by comparing to LPS stimulated cells as follows: not significant (ns), *** p

    Journal: Frontiers in Immunology

    Article Title: The Generation of an Engineered Interleukin-10 Protein With Improved Stability and Biological Function

    doi: 10.3389/fimmu.2020.01794

    Figure Lengend Snippet: Stable mouse IL-10 (STm) is biologically active in vitro . (A) Specific induction of STAT3 phosphorylation by STm (5 ng/mL) was verified by the use of spleen cells from different mouse lines: Wild type C57BL/6 (WT), IL-10 knockout (IL-10 −/− ) and IL-10 receptor knockout (IL-10R1 −/− ). (B) Luciferase activity was monitored as the area under the curve (AUC) from BMDMs reporter mouse; cells were either unstimulated or stimulated with LPS (10 ng/mL) in the presence or absence of IL-10 from HEK 293 EBNA supernatants. A commercial mouse IL-10 (CmIL-10) was used as a positive control, and un-transfected cells supernatants (Medium) served as a negative control. Significant difference considered by comparing to LPS stimulated cells as follows: not significant (ns), *** p

    Article Snippet: The nine amino acids (-G3 SG4 S-) and 13 amino acid linker(-G3 SG4 -SG4-) were generated by inserting the synthetic insert into the second monomer of mouse IL-10.

    Techniques: In Vitro, Knock-Out, Luciferase, Activity Assay, Positive Control, Transfection, Negative Control

    Biological stability of mouse IL-10 variants upon treatment at different temperatures and pH in vitro . (A) Area Under the Curve (AUC) of luciferase induction was measured after incubating 100 ng/mL Nm (black line) and STm (blue line) at 37°C in the time course. (B) Both Nm (black line) and STm (blue line) were treated at 55°C in time course before luciferase activity was measured as the area under the curve (AUC) after LPS-stimulated BMDMs of reporter mouse. (C–E) LPS-induced luciferase inhibition was measured after co-stimulated with either heat-treated at 55°C for 10 min (10 min) or untreated (0 min) of the commercial mouse IL-10 (CmIL-10), Nm and STm. CmIL-10 was used as a control in this experiment. The ED 50 was calculated as follow: (C) CmIL-10 heat-treated for (2.24 ng/mL) or untreated (0.137); (D) Nm heat-treated for (3.23 ng/mL) or untreated (0.8 ng/mL); (E) STm heat-treated for (0.09 ng/mL) or untreated (0.08 ng/mL). (F) Both Nm and STm at 100 ng/mL were pre-incubated with different pH buffers at 4°C for 24 h flowed by buffer exchange columns. BMDMs of the transgenic mouse then stimulated with LPS (10 ng/mL), and pH treated IL-10 (Nm and STm) at 10 ng/mL. The percentage of luciferase activities is relative to LPS treatment. The significant difference compared between pH treatments with neutral pH (pH7) of Nm and STm on LPS-stimulated cells *** p

    Journal: Frontiers in Immunology

    Article Title: The Generation of an Engineered Interleukin-10 Protein With Improved Stability and Biological Function

    doi: 10.3389/fimmu.2020.01794

    Figure Lengend Snippet: Biological stability of mouse IL-10 variants upon treatment at different temperatures and pH in vitro . (A) Area Under the Curve (AUC) of luciferase induction was measured after incubating 100 ng/mL Nm (black line) and STm (blue line) at 37°C in the time course. (B) Both Nm (black line) and STm (blue line) were treated at 55°C in time course before luciferase activity was measured as the area under the curve (AUC) after LPS-stimulated BMDMs of reporter mouse. (C–E) LPS-induced luciferase inhibition was measured after co-stimulated with either heat-treated at 55°C for 10 min (10 min) or untreated (0 min) of the commercial mouse IL-10 (CmIL-10), Nm and STm. CmIL-10 was used as a control in this experiment. The ED 50 was calculated as follow: (C) CmIL-10 heat-treated for (2.24 ng/mL) or untreated (0.137); (D) Nm heat-treated for (3.23 ng/mL) or untreated (0.8 ng/mL); (E) STm heat-treated for (0.09 ng/mL) or untreated (0.08 ng/mL). (F) Both Nm and STm at 100 ng/mL were pre-incubated with different pH buffers at 4°C for 24 h flowed by buffer exchange columns. BMDMs of the transgenic mouse then stimulated with LPS (10 ng/mL), and pH treated IL-10 (Nm and STm) at 10 ng/mL. The percentage of luciferase activities is relative to LPS treatment. The significant difference compared between pH treatments with neutral pH (pH7) of Nm and STm on LPS-stimulated cells *** p

    Article Snippet: The nine amino acids (-G3 SG4 S-) and 13 amino acid linker(-G3 SG4 -SG4-) were generated by inserting the synthetic insert into the second monomer of mouse IL-10.

    Techniques: In Vitro, Luciferase, Activity Assay, Inhibition, Incubation, Buffer Exchange, Transgenic Assay

    Mutations introduced into one monomer of stable mouse IL-10 at the IL-10R binding site (IL-10M2 Mu ). (A) The cDNA construct of both un-mutant IL-10 and IL-10M2 Mu was cloned into the expression vector (pCEP V19) and the cartoon illustration of IL-10M2 Mu with the location of the 4-points mutations at the IL-10 receptor-binding site on the helix A of IL-10 protein. (B) Purified IL-10M2 Mu detected by ELISA (1/2 dilutions) to evaluate the expression level. (C) Assessment of luciferase suppression after LPS (10 ng/mL) treatment and either co-treated with Nm (10 ng/mL), STm (10 ng/mL), or IL-10M2 Mu at (10 and 50 ng/mL). The significant difference is calculated concerning LPS stimulated cells only and symbolized as *** p

    Journal: Frontiers in Immunology

    Article Title: The Generation of an Engineered Interleukin-10 Protein With Improved Stability and Biological Function

    doi: 10.3389/fimmu.2020.01794

    Figure Lengend Snippet: Mutations introduced into one monomer of stable mouse IL-10 at the IL-10R binding site (IL-10M2 Mu ). (A) The cDNA construct of both un-mutant IL-10 and IL-10M2 Mu was cloned into the expression vector (pCEP V19) and the cartoon illustration of IL-10M2 Mu with the location of the 4-points mutations at the IL-10 receptor-binding site on the helix A of IL-10 protein. (B) Purified IL-10M2 Mu detected by ELISA (1/2 dilutions) to evaluate the expression level. (C) Assessment of luciferase suppression after LPS (10 ng/mL) treatment and either co-treated with Nm (10 ng/mL), STm (10 ng/mL), or IL-10M2 Mu at (10 and 50 ng/mL). The significant difference is calculated concerning LPS stimulated cells only and symbolized as *** p

    Article Snippet: The nine amino acids (-G3 SG4 S-) and 13 amino acid linker(-G3 SG4 -SG4-) were generated by inserting the synthetic insert into the second monomer of mouse IL-10.

    Techniques: Binding Assay, Construct, Mutagenesis, Clone Assay, Expressing, Plasmid Preparation, Purification, Enzyme-linked Immunosorbent Assay, Luciferase

    Suppression of LPS-induced dermal inflammation by Nm and STm. (A) Overview of healthy skin. Mouse skin consists of a 1–2-layer epidermis (Ep), which forms the hair follicles (Hf) and sebaceous glands (Td) by invagination in the dermis (De). The dermis consists of collagenous connective tissue. This is followed by the muscle layer of the Panniculus carnosus (Pc), the fatty tissue (Fg), and the trunk muscles (Rm). (B) Enlargement of healthy skin. The epithelium shows bluish cytoplasm and loose chromatin, as well as two intact hair follicles. (C) Overview of necrotic skin. In magnification, a large number of neutrophils is recognizable. (D) Enlargement of necrotic skin. The epithelial cells show reddish cytoplasm and condensed chromatin. The hair follicle is dead. (E) Overview skin section treated STm (2 μg) STm, which LPS was co-injected. (F) A table summarizing the effect of IL-10 (Nm/STm) at different concentrations on LPS-treated (10 μg) skin.

    Journal: Frontiers in Immunology

    Article Title: The Generation of an Engineered Interleukin-10 Protein With Improved Stability and Biological Function

    doi: 10.3389/fimmu.2020.01794

    Figure Lengend Snippet: Suppression of LPS-induced dermal inflammation by Nm and STm. (A) Overview of healthy skin. Mouse skin consists of a 1–2-layer epidermis (Ep), which forms the hair follicles (Hf) and sebaceous glands (Td) by invagination in the dermis (De). The dermis consists of collagenous connective tissue. This is followed by the muscle layer of the Panniculus carnosus (Pc), the fatty tissue (Fg), and the trunk muscles (Rm). (B) Enlargement of healthy skin. The epithelium shows bluish cytoplasm and loose chromatin, as well as two intact hair follicles. (C) Overview of necrotic skin. In magnification, a large number of neutrophils is recognizable. (D) Enlargement of necrotic skin. The epithelial cells show reddish cytoplasm and condensed chromatin. The hair follicle is dead. (E) Overview skin section treated STm (2 μg) STm, which LPS was co-injected. (F) A table summarizing the effect of IL-10 (Nm/STm) at different concentrations on LPS-treated (10 μg) skin.

    Article Snippet: The nine amino acids (-G3 SG4 S-) and 13 amino acid linker(-G3 SG4 -SG4-) were generated by inserting the synthetic insert into the second monomer of mouse IL-10.

    Techniques: Injection

    Expression and ELISA measurement of natural and stable mouse IL-10. (A) The cDNA construct of both natural mouse IL-10 (Nm) and stable mouse IL-10 (STm). The cartoon illustration of the expected protein folding of both Nm and STm versions. (B) The ribbon plot of the natural IL-10 homo-dimer with N and C terminus obtained from the previous study ( 13 ) and the molecular dynamics simulation of stable IL-10. The basis for this simulation is the crystal structure of the hIL-10 dimer ( 33 ) (PDB-ID: 2ILK). After about 2 ns of simulated time, the state of the system shown here was found. The peptide linker (-G3SG3-) is colored red. The molecular simulation of stable IL-10 was carried out by Jan-Philip Gehrcke (Biotechnology Center Dresden). (C) Both Nm and STm were stained with Coomassie Blue under reducing condition in SDS-page and (D) detected by IL-10 antibody in Western Blot. (E) Nm (black line) and STm proteins (blue line) were detected ELISA (1/2 dilutions). All data are representative of triplicate wells, and the bars represent standard error of the mean.

    Journal: Frontiers in Immunology

    Article Title: The Generation of an Engineered Interleukin-10 Protein With Improved Stability and Biological Function

    doi: 10.3389/fimmu.2020.01794

    Figure Lengend Snippet: Expression and ELISA measurement of natural and stable mouse IL-10. (A) The cDNA construct of both natural mouse IL-10 (Nm) and stable mouse IL-10 (STm). The cartoon illustration of the expected protein folding of both Nm and STm versions. (B) The ribbon plot of the natural IL-10 homo-dimer with N and C terminus obtained from the previous study ( 13 ) and the molecular dynamics simulation of stable IL-10. The basis for this simulation is the crystal structure of the hIL-10 dimer ( 33 ) (PDB-ID: 2ILK). After about 2 ns of simulated time, the state of the system shown here was found. The peptide linker (-G3SG3-) is colored red. The molecular simulation of stable IL-10 was carried out by Jan-Philip Gehrcke (Biotechnology Center Dresden). (C) Both Nm and STm were stained with Coomassie Blue under reducing condition in SDS-page and (D) detected by IL-10 antibody in Western Blot. (E) Nm (black line) and STm proteins (blue line) were detected ELISA (1/2 dilutions). All data are representative of triplicate wells, and the bars represent standard error of the mean.

    Article Snippet: The nine amino acids (-G3 SG4 S-) and 13 amino acid linker(-G3 SG4 -SG4-) were generated by inserting the synthetic insert into the second monomer of mouse IL-10.

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Construct, Staining, SDS Page, Western Blot