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  • 99
    Thermo Fisher interleukin 6
    Secretion of <t>interleukin-6</t> (IL-6), interleukin-8 (IL-8), and vascular endothelial growth factor A (VEGFA) from BM-MSCs after stimulation with sEV-OCS or sEV-OCS-Cis. ( A – C ) Secretion levels of ( A ) IL-6, ( B ) IL-8, and ( C ) VEGFA of BM-MSCs stimulated with sEV-OCS or sEV-OCS-Cis measured by multiplex fluorescent bead-based immunoassay analysis. BM-MSCs cell-cultured media (CM) was harvested 24 hours post-stimulation with sEV-OCS (CM MCS -sEV-OCS) or sEV-OCS-Cis (CM MCS -sEV-OCS-Cis). CM of BM-MSCs without sEV stimulation (CM MCS -non-sEV) was used as control. Results are mean ± SEM of n = 3. Statistical analysis was performed using the Kruskal–Wallis test followed by the Mann–Whitney test. * p
    Interleukin 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1698 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Millipore interleukin 6
    Concentrations of ( a ) IL-1β, ( b ) <t>IL-6,</t> ( c ) TNF-α ( d ) anti-OVA Ab in serum determined by ELISA. The results are representative of means ± SD ( n = 5; * p
    Interleukin 6, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 625 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson interleukin 6
    LPS-Trap-Fc1 to -Fc4 bind LPS and block LPS-mediated <t>IL-6</t> production in Mono Mac 6 cells. (A) Supernatants of HEK293T cells transfected with LPS-Trap-Fc1 were incubated in the presence (lane 1) or absence (lane 3) of biotin-LPS. Additionally, supernatants were preincubated with an anti-TLR4/MD-2 MAb (lane 2) or an excess of LPS (lane 4). Complexes were immunoprecipitated with streptavidin-Sepharose, subjected to SDS-PAGE, and analyzed by Western blotting with an anti-FLAG MAb. (B) RAW 264.7 cells were incubated with LPS-Trap-Fc1 or a medium control. Cells were stimulated with 100 ng/ml LPS overnight, and IL-6 levels in supernatants were determined by ELISA. *, P
    Interleukin 6, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 997 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    R&D Systems interleukin 6
    Niphatenone B enantiomers block transactivation of AR NTD. (A) Chemical structures of sintokamide, EPI-001, and niphatenone B enantiomers. (B) Transactivation assay of the AR NTD performed in LNCaP cells that were co-transfected with Gal4UAS-TATA-luciferase and AR-(1-558)-Gal4 DBD prior pre-treatment for 1 hour with 25 µM EPI-002, 7.0 µM niphatenones or DMSO vehicle control. Transactivation of AR NTD was induced by incubation with <t>IL-6</t> (50 ng/ml) for 24 hours. Luciferase activity was normalized to protein concentration. (C) Niphatenones inhibit the constitutively active AR V567es splice variant. Cos-1 cells co-transfected with PB-luciferase and an expression vector for ARvar567 were treated with each niphatenone B (1 µM) or DMSO vehicle control. After 24 hours of exposure, cells were harvested and luciferase activities were normalized to protein concentrations of the samples. Data represent the mean ± SEM of n = 3 separate experiments with triplicate wells. Student's t test: ** P
    Interleukin 6, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 3783 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher interleukin 10
    Effect of FTY720 on inflammatory cytokines, including monocyte chemoattractant protein-1 (MCP-1), macrophage-related TNF-α and IL-6, arginase-1 and <t>IL-10,</t> in the three groups. (A) Western blot analysis of MCP-1 and M1 and M2 macrophage-related
    Interleukin 10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 390 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    R&D Systems interleukin 4
    Inhibition of PI3Kγ impairs Akt phosphorylation and B cell migration in a transwell assay. a Inhibition of PI3Kγ or PI3Kδ significantly decreased the migration of JVM3 cells. JVM3 cells were cultured in medium or stimulated for 24 h with <t>CD40L/IL-4</t> and then incubated with the PI3Kγ-specific inhibitor CZC24832 (2 µM), the PI3Kδ - specific inhibitor idelalisib (1 µM), dual PI3Kδ/γ inhibitor duvelisib (1 µM), or the pan-PI3K inhibitor GDC0980 (1 µM) and subjected to a transwell migration assay. DMSO served as the vehicle control for the inhibitors, while SDF1α (100 ng/ml) served as the chemoattractant ( n = 7). b Effect of PI3Kγ inhibitor on Akt phosphorylation. JVM3 cells were pre-incubated with the indicated concentrations of CZC4832 (in μM) and then stimulated with SDF1α for 10 min. Akt Ser473 phosphorylation and total Akt levels were assessed by Western blot
    Interleukin 4, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1106 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson interleukin 2
    GMM production by mycobacteria cultured at a physiological glucose concentration. A, MAC was cultured in liquid media containing either 0.01 or 0.1% glucose, and the culture media were replaced with fresh media every day to maintain the glucose concentrations. After 5 days, the bacteria were harvested, and the total lipids were extracted. The methanol-insoluble fraction was then obtained from 100 μg of each total lipid preparation and analyzed by TLC. B, GMM-specific, CD1b-restricted TCR-expressing Jurkat T cells were cocultured with either C1R/CD1b or C1R/mock in the presence of different concentrations of the total lipids derived from the 0.1% glucose-containing ( upper panel ) and the 0.01% glucose-containing ( lower panel ) cultures. The T cell response was assessed by measuring <t>IL-2</t> released into the media.
    Interleukin 2, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 687 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore il 1β
    IL7AS and MIR3142HG regulate the <t>IL-1β-stimulated</t> inflammatory response in control fibroblasts. Control fibroblasts were transfected overnight with LNA antisense sequences against IL7AS (A/C/E/G) and MIR3142HG (B/D/F/H) or scrambled (negative) controls. Cell were then stimulated with IL-1β for 24 h prior to isolation of RNA and measurement of IL7AS (A) or MIR3142HG (B) by qRT-PCR or measurement of supernatant IL-6 (C,D) , IL-8 (E,F) , and CCL2 (G,H) by ELISA. Data represents the mean ± SEM of five control individuals. Following normalization against the IL1β-stimulated cells (100%), statistical significance was assessed (vs. IL1β-stimulated cells) using the repeat measures 1-way analysis of variance (ANOVA) with a Dunnett's test where * p
    Il 1β, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 5110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    PeproTech interleukin 4
    MPLA promotes the maturation of BMDCs via a TLR4-dependent pathway. Wild type (WT) and TLR4 knock-out (TLR4 −/− ) mice were euthanized, and their BM cells were collected from femurs and tibias for the culture of BMDCs in the presence of 20 ng/mL GM-CSF and 10 ng/mL <t>IL-4.</t> On day 7, BMDCs of three groups ( n = 4/group) were treated with DMEM (100 μL), MPLA (100 μL of 1 μg), and inactivated LBNSE (100 μL of 1 × 10 7 FFU), respectively. ( A ) Representative gating strategy for CD11c + CD86 + cells in BMDCs. ( B ) Representative flow cytometric plots of mature CD11c + CD86 + BMDCs from three groups. ( C ) Statistical results of CD11c + CD86 + BMDCs collected at 24 h post-stimulation. Data are presented as means ± SD. Asterisks indicate a significant difference between the groups at the levels of *** p
    Interleukin 4, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 669 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore interleukin 2
    Down‐regulation of proteins associated with cytotoxic activity of alloactivated peripheral blood mononuclear cells (PBMCs) by recombinant glycodelin (rGd). Cells isolated from the tumours excised on day 15 after injection of AP or APG were stained with human antibodies specific to CD8, interleukin <t>(IL)‐2,</t> eomesodermin (EOMES) and granzyme‐B. Protein expression was analysed using flow cytometry. Decrease in the protein expression in APG in comparison to AP is represented as the % inhibition. AP = alloactivated PBMCs; APG = alloactivated PBMCs treated with rGd.
    Interleukin 2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 429 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher interleukin il 1 β
    Change in pro-inflammatory cytokines on day 5 expressed as a percentage of baseline level (day 1). Significant differences in <t>interleukin-1β</t> (A) and interleukin-6 (B) levels were observed between groups. A trend towards significant difference
    Interleukin Il 1 β, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 340 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam interleukin 6
    Effect of SOG on serum fasting insulin, pancreatic insulin and pancreatic <t>IL-6</t> levels in experimental diabetic mice induced by STZ. The normal group represents mice without STZ injection and the model group represents mice with the induction of diabetes by STZ injection. Data are presented as the mean ± standard deviation (n=10 mice). **P
    Interleukin 6, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 352 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson interleukin 10
    IL4I1 inhibition of bacterial growth in vivo and associated plasmatic cytokine variations. ( A ) MSSA was added to HEK-PBS or IL4I1-PBS, and the mixes were injected intraperitoneally into groups of three C57Bl/6 mice (mean of 1.74×10 8 CFU/mouse). Twenty-four hours after injection, supernatants from dissociated spleens were serially diluted and inoculated onto LB agar plates. Bacterial colonies were then counted. The ratio of the CFU in each mouse, relative to the mean CFU from triplicate HEK-PBS mice from each of the four experiments was calculated. Data are represented as mean ratio ± SEM. ( B-C ) Interferon γ (IFNγ), tumor necrosis factor α (TNF), interleukin 6 (IL-6) and <t>interleukin</t> 10 (IL-10) were measured by ELISA in diluted plasma samples from groups of three mice injected with MSSA ( B ), or with 20 µg LPS ( C ). ELISA results are given as mean ± SEM from three experiments. * p
    Interleukin 10, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 466 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Novartis interleukin 2
    Influence of HDACi and DNMTi on NK susceptibility of MHH-CALL-4 cells . Leukemic cells were incubated with the indicated concentrations of HDACi and DNMTi for 48 h. Resting NK cells (A) or <t>IL-2</t> stimulated NK cells (B) were used as effector cells. A lysis-ratio was calculated from each experiment as following: specific lysis with HDACi/DNMTi/specific lysis without HDACi/DNMTi. Shown are mean values and standard deviation from an effector-to-target cell ratio of 20:1 [ n = 6 for vorinostat (four different donors), n = 15 for valproic acid (six different donors), n = 7 for decitabine (four different donors), n = 4 for azacytidine (four different donors), ** p
    Interleukin 2, supplied by Novartis, used in various techniques. Bioz Stars score: 92/100, based on 266 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher interleukin 2
    Effects of C 60 derivatives on T cell responses. a C 60 -P or C 60 (OH) 36 was added to responder splenocytes 30 min before the addition of stimulator splenocytes. After incubation of the cells for 4 days, the amount of <t>IL-2</t> in the supernatants was measured by ELISA. b One week after the third immunization with OVA and alum, mice were euthanized and single-cell suspensions of splenocytes were prepared. C 60 -P or C 60 (OH) 36 was added to the wells 30 min before the addition of OVA (100 μg/mL). After incubation of the cells for 3 days, the amount of IL-4 in the supernatants was measured by ELISA. c Purified CD4 + T cells were added to anti-CD3-coated plates. Each C 60 derivative or N -acetylcysteine (NAC) was added to the wells 30 min before the addition of anti-CD28. After incubation of the cells for 3 days, the amount of IL-2 in the supernatants was determined by ELISA. Data are means ± SDs for three to six independent cultures ( n = 3 to 6). * P
    Interleukin 2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 695 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    R&D Systems interleukin 3
    TBI selectively suppresses LKS + -cell replicative and clonogenic function. (A-F) Mice were exposed to 6.5 Gy TBI or not irradiated as control (CTL). At 28 days after TBI, BM-MNCs were isolated from irradiated and control mice. The clonogenic function of HPCs and HSCs was measured by CFC assay and day-35 CAFC assay, respectively. The frequencies of LKS - and LKS + cells in BM-MNCs were quantified by flow cytometry (B,E). The number of various CFUs and day-35 CAFCs is expressed as a function of BM-MNCs (A,D) or as a function of LKS - and LKS + cells, respectively (C,F). The data are presented as mean ± SE (n = 5 for CFC assay and n = 3 for CAFC assay). (G-H) At 28 days after TBI, BM-MNCs were isolated from irradiated and control mice and single LKS + cells were sorted into wells of rounded-bottom plates. After 14 days of culture with SCF/TPO, wells with a single seeded cell that had undergone at least one round of cell division were scored and graded by the number of cells produced. The results are presented as a percentage of LKS + cells that divided (G). Similarly, single sorted cells were cultured with <t>SCF/TPO/IL-3</t> for 14 days and the formation of hematopoietic-cell colonies ( > 50 cells) was scored based on the size of the colonies. The data are expressed as a percentage of LKS + cells that formed colony (H). * P
    Interleukin 3, supplied by R&D Systems, used in various techniques. Bioz Stars score: 97/100, based on 520 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    4Gene interleukin 4 gene expression
    Th2 differentiation driven by low concentration of peptide stimulation in vitro consists of an <t>IL-4–independent</t> initiation phase and an IL-4–dependent amplification phase . (A) TCR stimulation by low concentration of peptide induces IL-4–independent GATA-3 expression and IL-2–mediated Stat5 activation. (B) GATA-3 binds to CNS-1 and V A whereas activated Stat5 binds to HSII and HSIII of Il4 locus. Both are critical for TCR-mediated IL-4 production at the initial phase of Th2 cell differentiation. (C) IL-4 produced by T cells can further induce GATA-3 expression through Stat6 activation. GATA-3 also regulates itself once it reaches a certain threshold. Thus, IL-4–mediated GATA-3 expression together with IL-2–mediated Stat5 activation drives full Th2 differentiation. (D) High levels of GATA-3 and activated Stat5 play critical roles in inducing large amount of IL-4 production.
    Interleukin 4 Gene Expression, supplied by 4Gene, used in various techniques. Bioz Stars score: 89/100, based on 145 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher human il 6
    <t>IL-6/IL-8-arginase</t> I blockade reduces CD45 + CD33 low CD11b dim myeloid cell-mediated CD8 + T cell suppression. CFSE-labeled peripheral blood CD8 + T cells from healthy donors were co-cultured for 5 days with peripheral blood CD45 + CD33 low CD11b dim myeloid cells from GC patients in the presence or absence of 50% GC patient serum. A proportion of samples were either co-cultured in the presence or absence of IL-6 and/or IL-8 neutralizing abs and/or nor-NOHA, and the proportion of a IFN-γ expressing, b granzyme B expressing and c proliferating CD8 + T cells quantitated by flow cytometry. Data representative of three independent experiments ( n = 3). * p
    Human Il 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 580 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Bio-Rad interleukin 6
    Supplementation with  Lactobacillus reuteri  100-23 and  Lactobacillus gasseri  311476 mixture reduces inflammatory cytokines. A–F . Plasma levels of interleukin 4 (IL-4), interleukin 10 (IL-10), interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP-1), interleukin 8 (IL-8), and granulocyte colony-stimulating factor (G-CSF) in control mice (CT), in mice receiving lactobacilli (Lrg), in mice transplanted with BaF3 cells (BaF3) and in mice transplanted with BaF3 cells and receiving lactobacilli (BaF3-Lrg). Data with different superscript letters are significantly different (p
    Interleukin 6, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 307 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    R&D Systems interleukin 2
    (a) The percentage of blood T cells migrating within three-dimensional collagen gels after 6 days of <t>interleukin-2</t> (IL-2) stimulation compared with the unstimulated control ( n = 5, P
    Interleukin 2, supplied by R&D Systems, used in various techniques. Bioz Stars score: 97/100, based on 580 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Meso Scale Diagnostics LLC interleukin 6
    a - f Mitochondrial proteins and inflammatory cytokines levels in explant cultured media in SIM tissue and surrounding matched-normal tissue. Wilcoxon matched-pairs signed rank tests demonstrated significantly increased levels of a cytochrome c (n=12), b SMAC/Diablo (n=8), c IL-1beta (n=12), d <t>IL-6</t> (n=12), e IL-8 (n=12) and f TNF-alpha (n=12) in SIM tissue compared to surrounding normal epithelium. * p ≤ 0.05, ** p ≤ 0.005 and *** p ≤ 0.0005
    Interleukin 6, supplied by Meso Scale Diagnostics LLC, used in various techniques. Bioz Stars score: 93/100, based on 196 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson interleukin 4
    Th1 responses that are dominant over Th2 responses control L. donovani infection. The frequencies of CD4 + and CD8 + T cells producing IFN-γ and <t>IL-4</t> in the indicated groups were assessed using single-cell splenocyte suspensions left unstimulated or restimulated overnight with gp63 (5 μg/ml). (A) The frequencies of cells producing IFN-γ were assessed 10 days after vaccination both before and after 3 months of infection. lip, liposome encapsulation; Lip, empty liposomes. (B) The frequencies of cells producing IFN-γ were assessed 12 weeks after vaccination both before and after 3 months of infection. (C) Mice were challenged with L. donovani at 10 days, and the frequencies of cells producing IL-4 were determined at 3 months both before and after infectious challenge. (D) Frequencies of cells producing intracellular IL-4 were also determined in animals at 12 weeks postimmunization both before and after 3 months of infectious challenge. Data represent means ± standard errors of the means of the results obtained with three individual mice.
    Interleukin 4, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 301 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher mouse il 6
    Analysis of TLR signaling in Smcr8 −/− and Smcr8 I2T/I2T myeloid cells. ( A ) Peritoneal macrophages harvested from pedigrees R1418 [REF, Smcr8 +/+ ( n = 8); HET, Smcr8 +/p ( n = 15); VAR, Smcr8 p/p ( n = 2)] and R3960 [REF, Smcr8 +/+ ( n = 11); HET, Smcr8 +/p2 ( n = 18); VAR, Smcr8 p2/p2 ( n = 5)] were stimulated with 25 μg/mL of CpG. TNF in the culture medium as measured by ELISA 4 h later. ( B ) Manhattan plot showing P values of association between the phenotype of elevated TNF production in response to CpG and mutations identified in the two pedigrees in A calculated using a recessive model of inheritance. The −log 10 P values are plotted versus the chromosomal positions of mutations. Horizontal red and purple lines represent thresholds of P = 0.05 with or without Bonferroni correction, respectively. The P value for linkage of Smcr8 mutations with the elevated TNF production is indicated. ( C and D ) ELISA analysis of TNF secretion by peritoneal macrophages ( n = 5 or 6 mice per genotype) stimulated for 6 h with indicated concentrations of endosomal TLR ligands poly(I:C), R848, and CpG ( C ) or surface TLR ligands Pam3CSK4 and LPS ( D ). ( E and F ) ELISA analysis of <t>IL-6</t> secretion by BMDCs ( n = 4 mice per genotype) stimulated for 6 h with indicated concentrations of poly(I:C), R848, and CpG ( E ) or Pam3CSK4 and LPS ( F ). In A and C – F , data points represent individual mice. Data are representative of one ( A and B ) or three independent experiments ( C – F ). Data are expressed as means ± SD, and the significance of differences between genotypes was determined by one-way ANOVA with Dunnett’s multiple comparisons ( C and D ) or unpaired Student’s t test ( E and F ) (* P
    Mouse Il 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 553 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Secretion of interleukin-6 (IL-6), interleukin-8 (IL-8), and vascular endothelial growth factor A (VEGFA) from BM-MSCs after stimulation with sEV-OCS or sEV-OCS-Cis. ( A – C ) Secretion levels of ( A ) IL-6, ( B ) IL-8, and ( C ) VEGFA of BM-MSCs stimulated with sEV-OCS or sEV-OCS-Cis measured by multiplex fluorescent bead-based immunoassay analysis. BM-MSCs cell-cultured media (CM) was harvested 24 hours post-stimulation with sEV-OCS (CM MCS -sEV-OCS) or sEV-OCS-Cis (CM MCS -sEV-OCS-Cis). CM of BM-MSCs without sEV stimulation (CM MCS -non-sEV) was used as control. Results are mean ± SEM of n = 3. Statistical analysis was performed using the Kruskal–Wallis test followed by the Mann–Whitney test. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Small Extracellular Vesicles Released from Ovarian Cancer Spheroids in Response to Cisplatin Promote the Pro-Tumorigenic Activity of Mesenchymal Stem Cells

    doi: 10.3390/ijms20204972

    Figure Lengend Snippet: Secretion of interleukin-6 (IL-6), interleukin-8 (IL-8), and vascular endothelial growth factor A (VEGFA) from BM-MSCs after stimulation with sEV-OCS or sEV-OCS-Cis. ( A – C ) Secretion levels of ( A ) IL-6, ( B ) IL-8, and ( C ) VEGFA of BM-MSCs stimulated with sEV-OCS or sEV-OCS-Cis measured by multiplex fluorescent bead-based immunoassay analysis. BM-MSCs cell-cultured media (CM) was harvested 24 hours post-stimulation with sEV-OCS (CM MCS -sEV-OCS) or sEV-OCS-Cis (CM MCS -sEV-OCS-Cis). CM of BM-MSCs without sEV stimulation (CM MCS -non-sEV) was used as control. Results are mean ± SEM of n = 3. Statistical analysis was performed using the Kruskal–Wallis test followed by the Mann–Whitney test. * p

    Article Snippet: The concentrations of interleukin-6 (IL-6), interleukin-8 (IL-8), and vascular endothelial growth factor A (VEGFA) were determined in CM of BM-MSCs stimulated with sEV-OCS or sEV-OCS-Cis for 24 hours using a MAGPIX System (ThermoFisher Scientific, Waltham, MA, USA) as was described previously [ ].

    Techniques: Multiplex Assay, Bead-based Assay, Cell Culture, MANN-WHITNEY

    Concentrations of ( a ) IL-1β, ( b ) IL-6, ( c ) TNF-α ( d ) anti-OVA Ab in serum determined by ELISA. The results are representative of means ± SD ( n = 5; * p

    Journal: Polymers

    Article Title: Hydrogel is Superior to Fibrin Gel as Matrix of Stem Cells in Alleviating Antigen-Induced Arthritis

    doi: 10.3390/polym8050182

    Figure Lengend Snippet: Concentrations of ( a ) IL-1β, ( b ) IL-6, ( c ) TNF-α ( d ) anti-OVA Ab in serum determined by ELISA. The results are representative of means ± SD ( n = 5; * p

    Article Snippet: Percoll density gradient, thrombin, OVA and interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and anti-ovalbumin antibody (anti-OVA Ab) ELISA assay kits and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) were purchased from Sigma-Aldrich (Shanghai, China).

    Techniques: Enzyme-linked Immunosorbent Assay

    LPS-Trap-Fc1 to -Fc4 bind LPS and block LPS-mediated IL-6 production in Mono Mac 6 cells. (A) Supernatants of HEK293T cells transfected with LPS-Trap-Fc1 were incubated in the presence (lane 1) or absence (lane 3) of biotin-LPS. Additionally, supernatants were preincubated with an anti-TLR4/MD-2 MAb (lane 2) or an excess of LPS (lane 4). Complexes were immunoprecipitated with streptavidin-Sepharose, subjected to SDS-PAGE, and analyzed by Western blotting with an anti-FLAG MAb. (B) RAW 264.7 cells were incubated with LPS-Trap-Fc1 or a medium control. Cells were stimulated with 100 ng/ml LPS overnight, and IL-6 levels in supernatants were determined by ELISA. *, P

    Journal: Infection and Immunity

    Article Title: Lipopolysaccharide-Trap-Fc, a Multifunctional Agent To Battle Gram-Negative Bacteria ▿

    doi: 10.1128/IAI.00004-09

    Figure Lengend Snippet: LPS-Trap-Fc1 to -Fc4 bind LPS and block LPS-mediated IL-6 production in Mono Mac 6 cells. (A) Supernatants of HEK293T cells transfected with LPS-Trap-Fc1 were incubated in the presence (lane 1) or absence (lane 3) of biotin-LPS. Additionally, supernatants were preincubated with an anti-TLR4/MD-2 MAb (lane 2) or an excess of LPS (lane 4). Complexes were immunoprecipitated with streptavidin-Sepharose, subjected to SDS-PAGE, and analyzed by Western blotting with an anti-FLAG MAb. (B) RAW 264.7 cells were incubated with LPS-Trap-Fc1 or a medium control. Cells were stimulated with 100 ng/ml LPS overnight, and IL-6 levels in supernatants were determined by ELISA. *, P

    Article Snippet: The supernatants were tested for secreted interleukin-6 (IL-6) by ELISA (Becton Dickinson, Heidelberg, Germany).

    Techniques: Blocking Assay, Transfection, Incubation, Immunoprecipitation, SDS Page, Western Blot, Enzyme-linked Immunosorbent Assay

    Influence of erythropoietin on meningococcal-induced cytokine production. Interleukin (IL)-6 and tumour necrosis factor (TNF)-α-positive monocytes are shown in neonates ( n = 20), infants (defined as

    Journal:

    Article Title: Attenuation of monocyte proinflammatory cytokine responses to Neisseria meningitidis in children by erythropoietin

    doi: 10.1111/j.1365-2249.2008.03760.x

    Figure Lengend Snippet: Influence of erythropoietin on meningococcal-induced cytokine production. Interleukin (IL)-6 and tumour necrosis factor (TNF)-α-positive monocytes are shown in neonates ( n = 20), infants (defined as

    Article Snippet: Because non-haematopoietic effects of erythropoietin require higher concentrations [ ] we employed human recombinant erythropoietin beta (NeoRecormon, Firma Roche, Grenzen-Wylach, Germany), diluted in RPMI-1640 in final concentrations of 100 and 500 U/ml respectively, which was added to the cultures 1 h prior to stimulation with heat-inactivated meningococcal strains at a final concentration of 1·2 × 108 per ml for 5 h. As a positive control, lipopolysaccharide (LPS) obtained from Escherichia coli ( E. coli 0127: B8; no. L3129; Sigma, Deisenhofen, Germany) was added at a concentration of 30 ng/ml for 5 h. Cells were washed in Hanks's balanced salt solution (HBSS) and resuspended in a buffer consisting of HBSS, 0·1% saponin (Riedel de Haen, Hanover, Germany) and 0·01 M HEPES buffer (Seromed Biochrome, Berlin, Germany); 200 µl aliquots of cells were added to tubes containing 0·5 µg/10 µl of monoclonal antibodies (mAbs) against CD14, tumour necrosis factor (TNF)-α and interleukin (IL)-6 (Becton Dickinson, Heidelberg, Germany).

    Techniques:

    Niphatenone B enantiomers block transactivation of AR NTD. (A) Chemical structures of sintokamide, EPI-001, and niphatenone B enantiomers. (B) Transactivation assay of the AR NTD performed in LNCaP cells that were co-transfected with Gal4UAS-TATA-luciferase and AR-(1-558)-Gal4 DBD prior pre-treatment for 1 hour with 25 µM EPI-002, 7.0 µM niphatenones or DMSO vehicle control. Transactivation of AR NTD was induced by incubation with IL-6 (50 ng/ml) for 24 hours. Luciferase activity was normalized to protein concentration. (C) Niphatenones inhibit the constitutively active AR V567es splice variant. Cos-1 cells co-transfected with PB-luciferase and an expression vector for ARvar567 were treated with each niphatenone B (1 µM) or DMSO vehicle control. After 24 hours of exposure, cells were harvested and luciferase activities were normalized to protein concentrations of the samples. Data represent the mean ± SEM of n = 3 separate experiments with triplicate wells. Student's t test: ** P

    Journal: PLoS ONE

    Article Title: Characterization of Niphatenones that Inhibit Androgen Receptor N-Terminal Domain

    doi: 10.1371/journal.pone.0107991

    Figure Lengend Snippet: Niphatenone B enantiomers block transactivation of AR NTD. (A) Chemical structures of sintokamide, EPI-001, and niphatenone B enantiomers. (B) Transactivation assay of the AR NTD performed in LNCaP cells that were co-transfected with Gal4UAS-TATA-luciferase and AR-(1-558)-Gal4 DBD prior pre-treatment for 1 hour with 25 µM EPI-002, 7.0 µM niphatenones or DMSO vehicle control. Transactivation of AR NTD was induced by incubation with IL-6 (50 ng/ml) for 24 hours. Luciferase activity was normalized to protein concentration. (C) Niphatenones inhibit the constitutively active AR V567es splice variant. Cos-1 cells co-transfected with PB-luciferase and an expression vector for ARvar567 were treated with each niphatenone B (1 µM) or DMSO vehicle control. After 24 hours of exposure, cells were harvested and luciferase activities were normalized to protein concentrations of the samples. Data represent the mean ± SEM of n = 3 separate experiments with triplicate wells. Student's t test: ** P

    Article Snippet: Interleukin-6 was purchased from R & D Systems (Minneapolis, MN).

    Techniques: Blocking Assay, Transactivation Assay, Transfection, Luciferase, Incubation, Activity Assay, Protein Concentration, Variant Assay, Expressing, Plasmid Preparation

    Effect of FTY720 on inflammatory cytokines, including monocyte chemoattractant protein-1 (MCP-1), macrophage-related TNF-α and IL-6, arginase-1 and IL-10, in the three groups. (A) Western blot analysis of MCP-1 and M1 and M2 macrophage-related

    Journal: Acta Pharmacologica Sinica

    Article Title: FTY720 inhibits tubulointerstitial inflammation in albumin overload-induced nephropathy of rats via the Sphk1 pathway

    doi: 10.1038/aps.2014.100

    Figure Lengend Snippet: Effect of FTY720 on inflammatory cytokines, including monocyte chemoattractant protein-1 (MCP-1), macrophage-related TNF-α and IL-6, arginase-1 and IL-10, in the three groups. (A) Western blot analysis of MCP-1 and M1 and M2 macrophage-related

    Article Snippet: All real-time RT-PCR reactions were performed on ABI PRISM 7300 real-time PCR system (Applied Biosystems, CA, USA) using the SYBR Premix Ex Taq (Takara, Japan) with gene-specific primer sets of rat monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), and arginase-1 developed by Invitrogen (Invitrogen Biotechnology, Shanghai, China).

    Techniques: Western Blot

    Inhibition of PI3Kγ impairs Akt phosphorylation and B cell migration in a transwell assay. a Inhibition of PI3Kγ or PI3Kδ significantly decreased the migration of JVM3 cells. JVM3 cells were cultured in medium or stimulated for 24 h with CD40L/IL-4 and then incubated with the PI3Kγ-specific inhibitor CZC24832 (2 µM), the PI3Kδ - specific inhibitor idelalisib (1 µM), dual PI3Kδ/γ inhibitor duvelisib (1 µM), or the pan-PI3K inhibitor GDC0980 (1 µM) and subjected to a transwell migration assay. DMSO served as the vehicle control for the inhibitors, while SDF1α (100 ng/ml) served as the chemoattractant ( n = 7). b Effect of PI3Kγ inhibitor on Akt phosphorylation. JVM3 cells were pre-incubated with the indicated concentrations of CZC4832 (in μM) and then stimulated with SDF1α for 10 min. Akt Ser473 phosphorylation and total Akt levels were assessed by Western blot

    Journal: Leukemia

    Article Title: Distinct roles for phosphoinositide 3-kinases γ and δ in malignant B cell migration

    doi: 10.1038/s41375-018-0012-5

    Figure Lengend Snippet: Inhibition of PI3Kγ impairs Akt phosphorylation and B cell migration in a transwell assay. a Inhibition of PI3Kγ or PI3Kδ significantly decreased the migration of JVM3 cells. JVM3 cells were cultured in medium or stimulated for 24 h with CD40L/IL-4 and then incubated with the PI3Kγ-specific inhibitor CZC24832 (2 µM), the PI3Kδ - specific inhibitor idelalisib (1 µM), dual PI3Kδ/γ inhibitor duvelisib (1 µM), or the pan-PI3K inhibitor GDC0980 (1 µM) and subjected to a transwell migration assay. DMSO served as the vehicle control for the inhibitors, while SDF1α (100 ng/ml) served as the chemoattractant ( n = 7). b Effect of PI3Kγ inhibitor on Akt phosphorylation. JVM3 cells were pre-incubated with the indicated concentrations of CZC4832 (in μM) and then stimulated with SDF1α for 10 min. Akt Ser473 phosphorylation and total Akt levels were assessed by Western blot

    Article Snippet: CZC24832 has greater than 10-fold selectivity over PI3Kβ and greater than 100-fold selectivity over PI3Kα and PI3Kδ [ ]. α-IgM F(ab′)2 (Southern Biotech) was used at 10 μg/ml and CD40 ligand and interleukin 4 (R & D systems) were used at 50 ng/ml each.

    Techniques: Inhibition, Migration, Transwell Assay, Cell Culture, Incubation, Transwell Migration Assay, Western Blot

    Inhibition of PI3Kγ or PI3Kδ impairs CLL cell migration. a Effect of PI3Kγ inhibitor on CLL cell migration. CLL cells were cultured in medium or stimulated with CD40L+IL-4 for 24 h and subjected to a transwell migration assay. Graphs represent migration of cells from individual CLL patient samples with or without addition of PI3Kγ inhibitor CZC24832, connected by lines. b Inhibition of PI3Kγ or PI3Kδ decreases the migration of CLL cells to a similar extent, while dual PI3Kδ/γ inhibition using duvelisib has significantly greater effect than PI3Kγ inhibitor alone. c Combination of PI3Kγ inhibitor and PI3Kδ inhibitor decreases the migration of CLL cells to a greater extent than either inhibitor alone. Lines connect individual patient sample migration responses in the presence of the indicated inhibitors. Note all inhibitor-treated groups were significantly different than the control untreated group, whereas the CZC24832+idelalisib combination was not significantly different than duvelisib.

    Journal: Leukemia

    Article Title: Distinct roles for phosphoinositide 3-kinases γ and δ in malignant B cell migration

    doi: 10.1038/s41375-018-0012-5

    Figure Lengend Snippet: Inhibition of PI3Kγ or PI3Kδ impairs CLL cell migration. a Effect of PI3Kγ inhibitor on CLL cell migration. CLL cells were cultured in medium or stimulated with CD40L+IL-4 for 24 h and subjected to a transwell migration assay. Graphs represent migration of cells from individual CLL patient samples with or without addition of PI3Kγ inhibitor CZC24832, connected by lines. b Inhibition of PI3Kγ or PI3Kδ decreases the migration of CLL cells to a similar extent, while dual PI3Kδ/γ inhibition using duvelisib has significantly greater effect than PI3Kγ inhibitor alone. c Combination of PI3Kγ inhibitor and PI3Kδ inhibitor decreases the migration of CLL cells to a greater extent than either inhibitor alone. Lines connect individual patient sample migration responses in the presence of the indicated inhibitors. Note all inhibitor-treated groups were significantly different than the control untreated group, whereas the CZC24832+idelalisib combination was not significantly different than duvelisib.

    Article Snippet: CZC24832 has greater than 10-fold selectivity over PI3Kβ and greater than 100-fold selectivity over PI3Kα and PI3Kδ [ ]. α-IgM F(ab′)2 (Southern Biotech) was used at 10 μg/ml and CD40 ligand and interleukin 4 (R & D systems) were used at 50 ng/ml each.

    Techniques: Inhibition, Migration, Cell Culture, Transwell Migration Assay

    Expression of PI3Kγ subunits in malignant B cell lines and CLL patient samples. a , b CLL cells were stimulated with F(ab′) 2 α-IgM (10 µg/ml) or CD40L/IL-4 (50 ng/ml each) and harvested 24 h later for RNA extraction and RT-qPCR analysis. mRNA expression of a p110γ and b p101 were determined, and expression levels were normalized against the expression of TATA box-binding protein (TBP). Patients were divided into indolent vs. progressive groups based on IgVH mutation status. c Protein expression of p110γ and p101 in CLL samples in response to BCR stimulation or CD40L/IL-4 stimulation. Data are representative of nine patients analyzed

    Journal: Leukemia

    Article Title: Distinct roles for phosphoinositide 3-kinases γ and δ in malignant B cell migration

    doi: 10.1038/s41375-018-0012-5

    Figure Lengend Snippet: Expression of PI3Kγ subunits in malignant B cell lines and CLL patient samples. a , b CLL cells were stimulated with F(ab′) 2 α-IgM (10 µg/ml) or CD40L/IL-4 (50 ng/ml each) and harvested 24 h later for RNA extraction and RT-qPCR analysis. mRNA expression of a p110γ and b p101 were determined, and expression levels were normalized against the expression of TATA box-binding protein (TBP). Patients were divided into indolent vs. progressive groups based on IgVH mutation status. c Protein expression of p110γ and p101 in CLL samples in response to BCR stimulation or CD40L/IL-4 stimulation. Data are representative of nine patients analyzed

    Article Snippet: CZC24832 has greater than 10-fold selectivity over PI3Kβ and greater than 100-fold selectivity over PI3Kα and PI3Kδ [ ]. α-IgM F(ab′)2 (Southern Biotech) was used at 10 μg/ml and CD40 ligand and interleukin 4 (R & D systems) were used at 50 ng/ml each.

    Techniques: Expressing, RNA Extraction, Quantitative RT-PCR, Binding Assay, Mutagenesis

    GMM production by mycobacteria cultured at a physiological glucose concentration. A, MAC was cultured in liquid media containing either 0.01 or 0.1% glucose, and the culture media were replaced with fresh media every day to maintain the glucose concentrations. After 5 days, the bacteria were harvested, and the total lipids were extracted. The methanol-insoluble fraction was then obtained from 100 μg of each total lipid preparation and analyzed by TLC. B, GMM-specific, CD1b-restricted TCR-expressing Jurkat T cells were cocultured with either C1R/CD1b or C1R/mock in the presence of different concentrations of the total lipids derived from the 0.1% glucose-containing ( upper panel ) and the 0.01% glucose-containing ( lower panel ) cultures. The T cell response was assessed by measuring IL-2 released into the media.

    Journal: The Journal of Biological Chemistry

    Article Title: Mycolyltransferase-mediated Glycolipid Exchange in Mycobacteria *

    doi: 10.1074/jbc.M805776200

    Figure Lengend Snippet: GMM production by mycobacteria cultured at a physiological glucose concentration. A, MAC was cultured in liquid media containing either 0.01 or 0.1% glucose, and the culture media were replaced with fresh media every day to maintain the glucose concentrations. After 5 days, the bacteria were harvested, and the total lipids were extracted. The methanol-insoluble fraction was then obtained from 100 μg of each total lipid preparation and analyzed by TLC. B, GMM-specific, CD1b-restricted TCR-expressing Jurkat T cells were cocultured with either C1R/CD1b or C1R/mock in the presence of different concentrations of the total lipids derived from the 0.1% glucose-containing ( upper panel ) and the 0.01% glucose-containing ( lower panel ) cultures. The T cell response was assessed by measuring IL-2 released into the media.

    Article Snippet: After 20 h, aliquots of the culture supernatants were collected, and the amount of interleukin-2 (IL-2) released into the supernatants was measured by the IL-2 ELISA kit (BD Biosciences).

    Techniques: Cell Culture, Concentration Assay, Thin Layer Chromatography, Expressing, Derivative Assay

    IL7AS and MIR3142HG regulate the IL-1β-stimulated inflammatory response in control fibroblasts. Control fibroblasts were transfected overnight with LNA antisense sequences against IL7AS (A/C/E/G) and MIR3142HG (B/D/F/H) or scrambled (negative) controls. Cell were then stimulated with IL-1β for 24 h prior to isolation of RNA and measurement of IL7AS (A) or MIR3142HG (B) by qRT-PCR or measurement of supernatant IL-6 (C,D) , IL-8 (E,F) , and CCL2 (G,H) by ELISA. Data represents the mean ± SEM of five control individuals. Following normalization against the IL1β-stimulated cells (100%), statistical significance was assessed (vs. IL1β-stimulated cells) using the repeat measures 1-way analysis of variance (ANOVA) with a Dunnett's test where * p

    Journal: Frontiers in Immunology

    Article Title: Long Non-coding RNAs Are Central Regulators of the IL-1β-Induced Inflammatory Response in Normal and Idiopathic Pulmonary Lung Fibroblasts

    doi: 10.3389/fimmu.2018.02906

    Figure Lengend Snippet: IL7AS and MIR3142HG regulate the IL-1β-stimulated inflammatory response in control fibroblasts. Control fibroblasts were transfected overnight with LNA antisense sequences against IL7AS (A/C/E/G) and MIR3142HG (B/D/F/H) or scrambled (negative) controls. Cell were then stimulated with IL-1β for 24 h prior to isolation of RNA and measurement of IL7AS (A) or MIR3142HG (B) by qRT-PCR or measurement of supernatant IL-6 (C,D) , IL-8 (E,F) , and CCL2 (G,H) by ELISA. Data represents the mean ± SEM of five control individuals. Following normalization against the IL1β-stimulated cells (100%), statistical significance was assessed (vs. IL1β-stimulated cells) using the repeat measures 1-way analysis of variance (ANOVA) with a Dunnett's test where * p

    Article Snippet: On day 2, the cells were serum-starved with 1 ml of fresh medium (0.1% FBS) and treated with/without 3 ng/ml IL-1β (recombinant, expressed in E. coli , Sigma-Aldrich, I9401-5UG) and TPCA-1 (0, 0.1, 1, 10 μM) (T1452, Sigma-Aldrich) for 24 h before supernatants were collected and cells were harvested for RNA extraction.

    Techniques: Transfection, Isolation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    IL-1β-induced expression of IL7AS, MIR3142HG, and the inflammatory mediators is mediated via the NF-κB signaling pathway in control and IPF fibroblasts. Control and IPF fibroblasts were pre-incubated in the stated concentration of TPCA-1 or 0.1% (v/v) DMSO (vehicle) and then incubated in absence or presence of IL-1β for 24 h prior to measurement of IL6, IL8 and CCL2 release (A) and IL7AS and MIR3142HG expression (B) . Data is normalized against IL-1β stimulated cells (100%) and represents the mean ± SEM of five control and IPF patients. The logIC 50 for each individual was determined in GraphPad Prism and comparison between control and IPF groups was performed using an unpaired t -test. The IC 50 was calculated from the mean logIC 50 values.

    Journal: Frontiers in Immunology

    Article Title: Long Non-coding RNAs Are Central Regulators of the IL-1β-Induced Inflammatory Response in Normal and Idiopathic Pulmonary Lung Fibroblasts

    doi: 10.3389/fimmu.2018.02906

    Figure Lengend Snippet: IL-1β-induced expression of IL7AS, MIR3142HG, and the inflammatory mediators is mediated via the NF-κB signaling pathway in control and IPF fibroblasts. Control and IPF fibroblasts were pre-incubated in the stated concentration of TPCA-1 or 0.1% (v/v) DMSO (vehicle) and then incubated in absence or presence of IL-1β for 24 h prior to measurement of IL6, IL8 and CCL2 release (A) and IL7AS and MIR3142HG expression (B) . Data is normalized against IL-1β stimulated cells (100%) and represents the mean ± SEM of five control and IPF patients. The logIC 50 for each individual was determined in GraphPad Prism and comparison between control and IPF groups was performed using an unpaired t -test. The IC 50 was calculated from the mean logIC 50 values.

    Article Snippet: On day 2, the cells were serum-starved with 1 ml of fresh medium (0.1% FBS) and treated with/without 3 ng/ml IL-1β (recombinant, expressed in E. coli , Sigma-Aldrich, I9401-5UG) and TPCA-1 (0, 0.1, 1, 10 μM) (T1452, Sigma-Aldrich) for 24 h before supernatants were collected and cells were harvested for RNA extraction.

    Techniques: Expressing, Incubation, Concentration Assay

    Differential expression of mRNAs and lncRNAs following IL-1β-stimulation of control lung fibroblasts . (A) Heat map showing the differential expression of mRNAs in control fibroblasts following IL-1β stimulation for 6 h. (B) Pathway analysis of up-regulated mRNAs. (C) Top 10 most highly expressed lncRNA in non-stimulated control fibroblasts. (D) Heat map showing the differential expression of lncRNAs in control fibroblasts following IL-1β stimulation for 6 h.

    Journal: Frontiers in Immunology

    Article Title: Long Non-coding RNAs Are Central Regulators of the IL-1β-Induced Inflammatory Response in Normal and Idiopathic Pulmonary Lung Fibroblasts

    doi: 10.3389/fimmu.2018.02906

    Figure Lengend Snippet: Differential expression of mRNAs and lncRNAs following IL-1β-stimulation of control lung fibroblasts . (A) Heat map showing the differential expression of mRNAs in control fibroblasts following IL-1β stimulation for 6 h. (B) Pathway analysis of up-regulated mRNAs. (C) Top 10 most highly expressed lncRNA in non-stimulated control fibroblasts. (D) Heat map showing the differential expression of lncRNAs in control fibroblasts following IL-1β stimulation for 6 h.

    Article Snippet: On day 2, the cells were serum-starved with 1 ml of fresh medium (0.1% FBS) and treated with/without 3 ng/ml IL-1β (recombinant, expressed in E. coli , Sigma-Aldrich, I9401-5UG) and TPCA-1 (0, 0.1, 1, 10 μM) (T1452, Sigma-Aldrich) for 24 h before supernatants were collected and cells were harvested for RNA extraction.

    Techniques: Expressing

    IL7AS but not MIR3142HG regulates the IL-β-stimulated inflammatory response in IPF fibroblasts. IPF fibroblasts were transfected overnight with LNA antisense sequences against IL7AS (A,C,E,G) and MIR3142HG (B,D,F,H) or scrambled (negative) controls. Cell were then stimulated with IL-1β for 24 h prior to isolation of RNA and measurement of IL7AS (A) or MIR3142HG (B) by qRT-PCR or measurement of supernatant IL-6 (C,D) , IL-8 (E,F) , and CCL2 (G,H) by ELISA. Data represents the mean ± SEM of five IPF individuals. Following normalization against the IL1β-stimulated cells (100%), statistical significance was assessed (vs. IL1β-stimulated cells) using the repeat measures 1-way analysis of variance (ANOVA) with a Dunnett's test where * p

    Journal: Frontiers in Immunology

    Article Title: Long Non-coding RNAs Are Central Regulators of the IL-1β-Induced Inflammatory Response in Normal and Idiopathic Pulmonary Lung Fibroblasts

    doi: 10.3389/fimmu.2018.02906

    Figure Lengend Snippet: IL7AS but not MIR3142HG regulates the IL-β-stimulated inflammatory response in IPF fibroblasts. IPF fibroblasts were transfected overnight with LNA antisense sequences against IL7AS (A,C,E,G) and MIR3142HG (B,D,F,H) or scrambled (negative) controls. Cell were then stimulated with IL-1β for 24 h prior to isolation of RNA and measurement of IL7AS (A) or MIR3142HG (B) by qRT-PCR or measurement of supernatant IL-6 (C,D) , IL-8 (E,F) , and CCL2 (G,H) by ELISA. Data represents the mean ± SEM of five IPF individuals. Following normalization against the IL1β-stimulated cells (100%), statistical significance was assessed (vs. IL1β-stimulated cells) using the repeat measures 1-way analysis of variance (ANOVA) with a Dunnett's test where * p

    Article Snippet: On day 2, the cells were serum-starved with 1 ml of fresh medium (0.1% FBS) and treated with/without 3 ng/ml IL-1β (recombinant, expressed in E. coli , Sigma-Aldrich, I9401-5UG) and TPCA-1 (0, 0.1, 1, 10 μM) (T1452, Sigma-Aldrich) for 24 h before supernatants were collected and cells were harvested for RNA extraction.

    Techniques: Transfection, Isolation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    IL-1β-induced expression of IL7AS, MIR3142HG, miR-146a, miR-3142 and inflammatory mediators in control and IPF fibroblasts . (A) Control and IPF fibroblasts were incubated in the absence of presence of IL-1β for 24 h and the fold-change in the IL7AS and MIR3142 expression determined by qRT-PCR, (B) ). Control and IPF fibroblasts were incubated in the absence of presence of IL-1β for 24 h before determination of the fold-change in miR-3142 and miR-146a expression by qRT-PCR (C) and the release of IL-6, IL-8, and CCL2 by ELISA (D) . Values are the mean ± SEM of five control and IPF patients and statistical significance was assessed using 1-way analysis of variance (ANOVA) where * p

    Journal: Frontiers in Immunology

    Article Title: Long Non-coding RNAs Are Central Regulators of the IL-1β-Induced Inflammatory Response in Normal and Idiopathic Pulmonary Lung Fibroblasts

    doi: 10.3389/fimmu.2018.02906

    Figure Lengend Snippet: IL-1β-induced expression of IL7AS, MIR3142HG, miR-146a, miR-3142 and inflammatory mediators in control and IPF fibroblasts . (A) Control and IPF fibroblasts were incubated in the absence of presence of IL-1β for 24 h and the fold-change in the IL7AS and MIR3142 expression determined by qRT-PCR, (B) ). Control and IPF fibroblasts were incubated in the absence of presence of IL-1β for 24 h before determination of the fold-change in miR-3142 and miR-146a expression by qRT-PCR (C) and the release of IL-6, IL-8, and CCL2 by ELISA (D) . Values are the mean ± SEM of five control and IPF patients and statistical significance was assessed using 1-way analysis of variance (ANOVA) where * p

    Article Snippet: On day 2, the cells were serum-starved with 1 ml of fresh medium (0.1% FBS) and treated with/without 3 ng/ml IL-1β (recombinant, expressed in E. coli , Sigma-Aldrich, I9401-5UG) and TPCA-1 (0, 0.1, 1, 10 μM) (T1452, Sigma-Aldrich) for 24 h before supernatants were collected and cells were harvested for RNA extraction.

    Techniques: Expressing, Incubation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    MPLA promotes the maturation of BMDCs via a TLR4-dependent pathway. Wild type (WT) and TLR4 knock-out (TLR4 −/− ) mice were euthanized, and their BM cells were collected from femurs and tibias for the culture of BMDCs in the presence of 20 ng/mL GM-CSF and 10 ng/mL IL-4. On day 7, BMDCs of three groups ( n = 4/group) were treated with DMEM (100 μL), MPLA (100 μL of 1 μg), and inactivated LBNSE (100 μL of 1 × 10 7 FFU), respectively. ( A ) Representative gating strategy for CD11c + CD86 + cells in BMDCs. ( B ) Representative flow cytometric plots of mature CD11c + CD86 + BMDCs from three groups. ( C ) Statistical results of CD11c + CD86 + BMDCs collected at 24 h post-stimulation. Data are presented as means ± SD. Asterisks indicate a significant difference between the groups at the levels of *** p

    Journal: Viruses

    Article Title: Monophosphoryl-Lipid A (MPLA) is an Efficacious Adjuvant for Inactivated Rabies Vaccines

    doi: 10.3390/v11121118

    Figure Lengend Snippet: MPLA promotes the maturation of BMDCs via a TLR4-dependent pathway. Wild type (WT) and TLR4 knock-out (TLR4 −/− ) mice were euthanized, and their BM cells were collected from femurs and tibias for the culture of BMDCs in the presence of 20 ng/mL GM-CSF and 10 ng/mL IL-4. On day 7, BMDCs of three groups ( n = 4/group) were treated with DMEM (100 μL), MPLA (100 μL of 1 μg), and inactivated LBNSE (100 μL of 1 × 10 7 FFU), respectively. ( A ) Representative gating strategy for CD11c + CD86 + cells in BMDCs. ( B ) Representative flow cytometric plots of mature CD11c + CD86 + BMDCs from three groups. ( C ) Statistical results of CD11c + CD86 + BMDCs collected at 24 h post-stimulation. Data are presented as means ± SD. Asterisks indicate a significant difference between the groups at the levels of *** p

    Article Snippet: Wild type (WT) or TLR4 knock-out (TLR4−/− ) mice-derived bone marrow-derived dendritic cells (BMDC) were maintained in RMPI-1640 medium supplemented with 20 ng/mL of granulocyte-macrophage colony stimulating factor (GM-CSF, cat. no. 315-03-20) and 10 ng/mL of interleukin-4 (IL-4, cat. no. 214-14-20) purchased from Peprotech Inc. (Rocky Hill, NJ, USA), and BMDC cells were utilized to detect TLR4-dependent cell activation.

    Techniques: Knock-Out, Mouse Assay, Flow Cytometry

    Down‐regulation of proteins associated with cytotoxic activity of alloactivated peripheral blood mononuclear cells (PBMCs) by recombinant glycodelin (rGd). Cells isolated from the tumours excised on day 15 after injection of AP or APG were stained with human antibodies specific to CD8, interleukin (IL)‐2, eomesodermin (EOMES) and granzyme‐B. Protein expression was analysed using flow cytometry. Decrease in the protein expression in APG in comparison to AP is represented as the % inhibition. AP = alloactivated PBMCs; APG = alloactivated PBMCs treated with rGd.

    Journal: Clinical and Experimental Immunology

    Article Title: Immunomodulatory activity of glycodelin: implications in allograft rejection

    doi: 10.1111/cei.13096

    Figure Lengend Snippet: Down‐regulation of proteins associated with cytotoxic activity of alloactivated peripheral blood mononuclear cells (PBMCs) by recombinant glycodelin (rGd). Cells isolated from the tumours excised on day 15 after injection of AP or APG were stained with human antibodies specific to CD8, interleukin (IL)‐2, eomesodermin (EOMES) and granzyme‐B. Protein expression was analysed using flow cytometry. Decrease in the protein expression in APG in comparison to AP is represented as the % inhibition. AP = alloactivated PBMCs; APG = alloactivated PBMCs treated with rGd.

    Article Snippet: PBMCs were cultured with HepG2e cells at a ratio of 20 : 1 for 96 h in Iscove's modified Dulbecco's medium (IMDM; Sigma‐Aldrich) supplemented with 15% FBS, 1 mM glutamax; 5 U/ml of interleukin‐2 (IL‐2; Sigma‐Aldrich) was added to the culture at 0 and 48 h.

    Techniques: Activity Assay, Recombinant, Isolation, Injection, Staining, Expressing, Flow Cytometry, Cytometry, Inhibition

    Down‐regulation of genes associated with cytotoxic activity of alloactivated peripheral blood mononuclear cells (PBMCs) by recombinant glycodelin (rGd). RNA was isolated from the tumours after injection of AP or APG. Real‐time polymerase chain reaction (PCR) was performed to detect the mRNA levels of human (a) CD4, (b) CD8, (c) interleukin (IL)‐2, (d) granzyme‐B, (e) eomesodermin (EOMES), (f) tumour necrosis factor (TNF)‐α and (g) IL‐6. The inset table tabulates the fold change in mRNA levels of each protein in APG in comparison to AP on day 3. Data are represented as the fold expression relative to the day 3 AP for each protein (Student's t ‐test; * P

    Journal: Clinical and Experimental Immunology

    Article Title: Immunomodulatory activity of glycodelin: implications in allograft rejection

    doi: 10.1111/cei.13096

    Figure Lengend Snippet: Down‐regulation of genes associated with cytotoxic activity of alloactivated peripheral blood mononuclear cells (PBMCs) by recombinant glycodelin (rGd). RNA was isolated from the tumours after injection of AP or APG. Real‐time polymerase chain reaction (PCR) was performed to detect the mRNA levels of human (a) CD4, (b) CD8, (c) interleukin (IL)‐2, (d) granzyme‐B, (e) eomesodermin (EOMES), (f) tumour necrosis factor (TNF)‐α and (g) IL‐6. The inset table tabulates the fold change in mRNA levels of each protein in APG in comparison to AP on day 3. Data are represented as the fold expression relative to the day 3 AP for each protein (Student's t ‐test; * P

    Article Snippet: PBMCs were cultured with HepG2e cells at a ratio of 20 : 1 for 96 h in Iscove's modified Dulbecco's medium (IMDM; Sigma‐Aldrich) supplemented with 15% FBS, 1 mM glutamax; 5 U/ml of interleukin‐2 (IL‐2; Sigma‐Aldrich) was added to the culture at 0 and 48 h.

    Techniques: Activity Assay, Recombinant, Isolation, Injection, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Expressing

    Change in pro-inflammatory cytokines on day 5 expressed as a percentage of baseline level (day 1). Significant differences in interleukin-1β (A) and interleukin-6 (B) levels were observed between groups. A trend towards significant difference

    Journal: Critical care medicine

    Article Title: Effect of cholecalciferol supplementation on vitamin D status and cathelicidin levels in sepsis: A randomized, placebo-controlled trial

    doi: 10.1097/CCM.0000000000001148

    Figure Lengend Snippet: Change in pro-inflammatory cytokines on day 5 expressed as a percentage of baseline level (day 1). Significant differences in interleukin-1β (A) and interleukin-6 (B) levels were observed between groups. A trend towards significant difference

    Article Snippet: Plasma cytokine levels of interleukin (IL)-1β, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ were measured using a commercially available kit (Cytokine Human Magnetic 10-Plex Panel, Life Technologies, Grand Island, NY) using a Luminex FLEXMAP 3D (Luminex, Austin, TX) multiplex assay analyzer.

    Techniques:

    Resolvin D1 restores the resolution of corneal inflammation. A : Hematoxylin and eosin (H E) staining showed the representative histologic appearance of the healing cornea 24 h after removal of the corneal epithelium in the control, diabetic, and resolvin D1 (RvD1)-treated diabetic mice. B : Corneas harvested 24 h after injury were homogenized and assayed for levels of myeloperoxidase (MPO) activity and tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) expression with enzyme-linked immunosorbent assay (ELISA; n=6). C : Immunofluorescence staining was performed with the macrophage marker anti-F4/80 (green fluorescence) and the M2 macrophage marker anti-CD206 (red fluorescence) 48 h after removal of the corneal epithelium. Data are given as the mean ± standard deviation (SD); *p

    Journal: Molecular Vision

    Article Title: Resolvin D1 promotes corneal epithelial wound healing and restoration of mechanical sensation in diabetic mice

    doi:

    Figure Lengend Snippet: Resolvin D1 restores the resolution of corneal inflammation. A : Hematoxylin and eosin (H E) staining showed the representative histologic appearance of the healing cornea 24 h after removal of the corneal epithelium in the control, diabetic, and resolvin D1 (RvD1)-treated diabetic mice. B : Corneas harvested 24 h after injury were homogenized and assayed for levels of myeloperoxidase (MPO) activity and tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) expression with enzyme-linked immunosorbent assay (ELISA; n=6). C : Immunofluorescence staining was performed with the macrophage marker anti-F4/80 (green fluorescence) and the M2 macrophage marker anti-CD206 (red fluorescence) 48 h after removal of the corneal epithelium. Data are given as the mean ± standard deviation (SD); *p

    Article Snippet: The supernatants were subjected to quantitative sandwich immunoassay with enzyme-linked immunosorbent assay (ELISA) kits, including myeloperoxidase (MPO; USCN, Wuhan, China), interleukin-1 beta (IL-1β, eBioscience, San Diego, CA), and tumor necrosis factor alpha (TNF-α, R & D Systems, Minneapolis, MN) according to the manufacturers’ procedures.

    Techniques: Staining, Mouse Assay, Activity Assay, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Marker, Fluorescence, Standard Deviation

    Effect of SOG on serum fasting insulin, pancreatic insulin and pancreatic IL-6 levels in experimental diabetic mice induced by STZ. The normal group represents mice without STZ injection and the model group represents mice with the induction of diabetes by STZ injection. Data are presented as the mean ± standard deviation (n=10 mice). **P

    Journal: Molecular Medicine Reports

    Article Title: Syringaresinol-di-O-β-D-glucoside, a phenolic compound from Polygonatum sibiricum, exhibits an antidiabetic and antioxidative effect on a streptozotocin-induced mouse model of diabetes

    doi: 10.3892/mmr.2018.9580

    Figure Lengend Snippet: Effect of SOG on serum fasting insulin, pancreatic insulin and pancreatic IL-6 levels in experimental diabetic mice induced by STZ. The normal group represents mice without STZ injection and the model group represents mice with the induction of diabetes by STZ injection. Data are presented as the mean ± standard deviation (n=10 mice). **P

    Article Snippet: Insulin (cat. no. ab100578) and interleukin-6 (IL-6; cat. no. ab100712) ELISA kits, and transforming growth factor-β1 (TGF-β1; cat. no. ab9758) and GAPDH (cat. no. ab8245) antibodies, were purchased from Abcam (Cambridge, MA, USA).

    Techniques: Mouse Assay, Injection, Standard Deviation

    IL4I1 inhibition of bacterial growth in vivo and associated plasmatic cytokine variations. ( A ) MSSA was added to HEK-PBS or IL4I1-PBS, and the mixes were injected intraperitoneally into groups of three C57Bl/6 mice (mean of 1.74×10 8 CFU/mouse). Twenty-four hours after injection, supernatants from dissociated spleens were serially diluted and inoculated onto LB agar plates. Bacterial colonies were then counted. The ratio of the CFU in each mouse, relative to the mean CFU from triplicate HEK-PBS mice from each of the four experiments was calculated. Data are represented as mean ratio ± SEM. ( B-C ) Interferon γ (IFNγ), tumor necrosis factor α (TNF), interleukin 6 (IL-6) and interleukin 10 (IL-10) were measured by ELISA in diluted plasma samples from groups of three mice injected with MSSA ( B ), or with 20 µg LPS ( C ). ELISA results are given as mean ± SEM from three experiments. * p

    Journal: PLoS ONE

    Article Title: Antibacterial Properties of the Mammalian L-Amino Acid Oxidase IL4I1

    doi: 10.1371/journal.pone.0054589

    Figure Lengend Snippet: IL4I1 inhibition of bacterial growth in vivo and associated plasmatic cytokine variations. ( A ) MSSA was added to HEK-PBS or IL4I1-PBS, and the mixes were injected intraperitoneally into groups of three C57Bl/6 mice (mean of 1.74×10 8 CFU/mouse). Twenty-four hours after injection, supernatants from dissociated spleens were serially diluted and inoculated onto LB agar plates. Bacterial colonies were then counted. The ratio of the CFU in each mouse, relative to the mean CFU from triplicate HEK-PBS mice from each of the four experiments was calculated. Data are represented as mean ratio ± SEM. ( B-C ) Interferon γ (IFNγ), tumor necrosis factor α (TNF), interleukin 6 (IL-6) and interleukin 10 (IL-10) were measured by ELISA in diluted plasma samples from groups of three mice injected with MSSA ( B ), or with 20 µg LPS ( C ). ELISA results are given as mean ± SEM from three experiments. * p

    Article Snippet: ELISA Cytokines interferon γ (IFNγ), tumor necrosis factor α (TNF), interleukin 6 (IL-6) and interleukin 10 (IL-10) were detected in mouse plasma samples diluted four fold in sample diluent by ELISA according to the manufacturer’s instructions (BD Biosciences, Le Pont de Claix, France).

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

    Influence of HDACi and DNMTi on NK susceptibility of MHH-CALL-4 cells . Leukemic cells were incubated with the indicated concentrations of HDACi and DNMTi for 48 h. Resting NK cells (A) or IL-2 stimulated NK cells (B) were used as effector cells. A lysis-ratio was calculated from each experiment as following: specific lysis with HDACi/DNMTi/specific lysis without HDACi/DNMTi. Shown are mean values and standard deviation from an effector-to-target cell ratio of 20:1 [ n = 6 for vorinostat (four different donors), n = 15 for valproic acid (six different donors), n = 7 for decitabine (four different donors), n = 4 for azacytidine (four different donors), ** p

    Journal: Frontiers in Oncology

    Article Title: Influence of Histone Deacetylase Inhibitors and DNA-Methyltransferase Inhibitors on the NK Cell-Mediated Lysis of Pediatric B-Lineage Leukemia

    doi: 10.3389/fonc.2013.00099

    Figure Lengend Snippet: Influence of HDACi and DNMTi on NK susceptibility of MHH-CALL-4 cells . Leukemic cells were incubated with the indicated concentrations of HDACi and DNMTi for 48 h. Resting NK cells (A) or IL-2 stimulated NK cells (B) were used as effector cells. A lysis-ratio was calculated from each experiment as following: specific lysis with HDACi/DNMTi/specific lysis without HDACi/DNMTi. Shown are mean values and standard deviation from an effector-to-target cell ratio of 20:1 [ n = 6 for vorinostat (four different donors), n = 15 for valproic acid (six different donors), n = 7 for decitabine (four different donors), n = 4 for azacytidine (four different donors), ** p

    Article Snippet: For expansion PMNC were incubated with irradiated (100 Gy) K562mb15-41BBL cells at a ratio of 1:1.5 in RPMI 1640 supplemented with 10% human AB-serum, l -glutamine, and 100 IU/ml interleukin-2 (Proleukin, Novartis, Basel, Suisse).

    Techniques: Incubation, Lysis, Standard Deviation

    Effects of C 60 derivatives on T cell responses. a C 60 -P or C 60 (OH) 36 was added to responder splenocytes 30 min before the addition of stimulator splenocytes. After incubation of the cells for 4 days, the amount of IL-2 in the supernatants was measured by ELISA. b One week after the third immunization with OVA and alum, mice were euthanized and single-cell suspensions of splenocytes were prepared. C 60 -P or C 60 (OH) 36 was added to the wells 30 min before the addition of OVA (100 μg/mL). After incubation of the cells for 3 days, the amount of IL-4 in the supernatants was measured by ELISA. c Purified CD4 + T cells were added to anti-CD3-coated plates. Each C 60 derivative or N -acetylcysteine (NAC) was added to the wells 30 min before the addition of anti-CD28. After incubation of the cells for 3 days, the amount of IL-2 in the supernatants was determined by ELISA. Data are means ± SDs for three to six independent cultures ( n = 3 to 6). * P

    Journal: Nanoscale Research Letters

    Article Title: Potential Suppressive Effects of Two C60 Fullerene Derivatives on Acquired Immunity

    doi: 10.1186/s11671-016-1663-7

    Figure Lengend Snippet: Effects of C 60 derivatives on T cell responses. a C 60 -P or C 60 (OH) 36 was added to responder splenocytes 30 min before the addition of stimulator splenocytes. After incubation of the cells for 4 days, the amount of IL-2 in the supernatants was measured by ELISA. b One week after the third immunization with OVA and alum, mice were euthanized and single-cell suspensions of splenocytes were prepared. C 60 -P or C 60 (OH) 36 was added to the wells 30 min before the addition of OVA (100 μg/mL). After incubation of the cells for 3 days, the amount of IL-4 in the supernatants was measured by ELISA. c Purified CD4 + T cells were added to anti-CD3-coated plates. Each C 60 derivative or N -acetylcysteine (NAC) was added to the wells 30 min before the addition of anti-CD28. After incubation of the cells for 3 days, the amount of IL-2 in the supernatants was determined by ELISA. Data are means ± SDs for three to six independent cultures ( n = 3 to 6). * P

    Article Snippet: After incubation of the cells for 4 days at 37 °C (95 % room air, 5 % CO2 ), the amount of interleukin 2 (IL-2) released into an aliquot of culture supernatant was measured with a murine IL-2 enzyme-linked immunosorbent assay (ELISA) kit (eBioscience, San Diego, CA, USA) in accordance with the manufacturer’s instructions.

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

    TBI selectively suppresses LKS + -cell replicative and clonogenic function. (A-F) Mice were exposed to 6.5 Gy TBI or not irradiated as control (CTL). At 28 days after TBI, BM-MNCs were isolated from irradiated and control mice. The clonogenic function of HPCs and HSCs was measured by CFC assay and day-35 CAFC assay, respectively. The frequencies of LKS - and LKS + cells in BM-MNCs were quantified by flow cytometry (B,E). The number of various CFUs and day-35 CAFCs is expressed as a function of BM-MNCs (A,D) or as a function of LKS - and LKS + cells, respectively (C,F). The data are presented as mean ± SE (n = 5 for CFC assay and n = 3 for CAFC assay). (G-H) At 28 days after TBI, BM-MNCs were isolated from irradiated and control mice and single LKS + cells were sorted into wells of rounded-bottom plates. After 14 days of culture with SCF/TPO, wells with a single seeded cell that had undergone at least one round of cell division were scored and graded by the number of cells produced. The results are presented as a percentage of LKS + cells that divided (G). Similarly, single sorted cells were cultured with SCF/TPO/IL-3 for 14 days and the formation of hematopoietic-cell colonies ( > 50 cells) was scored based on the size of the colonies. The data are expressed as a percentage of LKS + cells that formed colony (H). * P

    Journal: Blood

    Article Title: Total body irradiation selectively induces murine hematopoietic stem cell senescence

    doi: 10.1182/blood-2005-04-1418

    Figure Lengend Snippet: TBI selectively suppresses LKS + -cell replicative and clonogenic function. (A-F) Mice were exposed to 6.5 Gy TBI or not irradiated as control (CTL). At 28 days after TBI, BM-MNCs were isolated from irradiated and control mice. The clonogenic function of HPCs and HSCs was measured by CFC assay and day-35 CAFC assay, respectively. The frequencies of LKS - and LKS + cells in BM-MNCs were quantified by flow cytometry (B,E). The number of various CFUs and day-35 CAFCs is expressed as a function of BM-MNCs (A,D) or as a function of LKS - and LKS + cells, respectively (C,F). The data are presented as mean ± SE (n = 5 for CFC assay and n = 3 for CAFC assay). (G-H) At 28 days after TBI, BM-MNCs were isolated from irradiated and control mice and single LKS + cells were sorted into wells of rounded-bottom plates. After 14 days of culture with SCF/TPO, wells with a single seeded cell that had undergone at least one round of cell division were scored and graded by the number of cells produced. The results are presented as a percentage of LKS + cells that divided (G). Similarly, single sorted cells were cultured with SCF/TPO/IL-3 for 14 days and the formation of hematopoietic-cell colonies ( > 50 cells) was scored based on the size of the colonies. The data are expressed as a percentage of LKS + cells that formed colony (H). * P

    Article Snippet: Recombinant mouse thrombopoietin (TPO), stem-cell factor (SCF), and interleukin-3 (IL-3) were purchased from R & D Systems (Minneapolis, MN).

    Techniques: Mouse Assay, Irradiation, CTL Assay, Isolation, Flow Cytometry, Cytometry, Produced, Cell Culture

    Th2 differentiation driven by low concentration of peptide stimulation in vitro consists of an IL-4–independent initiation phase and an IL-4–dependent amplification phase . (A) TCR stimulation by low concentration of peptide induces IL-4–independent GATA-3 expression and IL-2–mediated Stat5 activation. (B) GATA-3 binds to CNS-1 and V A whereas activated Stat5 binds to HSII and HSIII of Il4 locus. Both are critical for TCR-mediated IL-4 production at the initial phase of Th2 cell differentiation. (C) IL-4 produced by T cells can further induce GATA-3 expression through Stat6 activation. GATA-3 also regulates itself once it reaches a certain threshold. Thus, IL-4–mediated GATA-3 expression together with IL-2–mediated Stat5 activation drives full Th2 differentiation. (D) High levels of GATA-3 and activated Stat5 play critical roles in inducing large amount of IL-4 production.

    Journal: Blood

    Article Title: CD4 T cells: fates, functions, and faults

    doi: 10.1182/blood-2008-05-078154

    Figure Lengend Snippet: Th2 differentiation driven by low concentration of peptide stimulation in vitro consists of an IL-4–independent initiation phase and an IL-4–dependent amplification phase . (A) TCR stimulation by low concentration of peptide induces IL-4–independent GATA-3 expression and IL-2–mediated Stat5 activation. (B) GATA-3 binds to CNS-1 and V A whereas activated Stat5 binds to HSII and HSIII of Il4 locus. Both are critical for TCR-mediated IL-4 production at the initial phase of Th2 cell differentiation. (C) IL-4 produced by T cells can further induce GATA-3 expression through Stat6 activation. GATA-3 also regulates itself once it reaches a certain threshold. Thus, IL-4–mediated GATA-3 expression together with IL-2–mediated Stat5 activation drives full Th2 differentiation. (D) High levels of GATA-3 and activated Stat5 play critical roles in inducing large amount of IL-4 production.

    Article Snippet: Interferon regulatory factor 4 (IRF4) interacts with NFATc2 to modulate interleukin 4 gene expression.

    Techniques: Concentration Assay, In Vitro, Amplification, Expressing, Activation Assay, Cell Differentiation, Produced

    IL-6/IL-8-arginase I blockade reduces CD45 + CD33 low CD11b dim myeloid cell-mediated CD8 + T cell suppression. CFSE-labeled peripheral blood CD8 + T cells from healthy donors were co-cultured for 5 days with peripheral blood CD45 + CD33 low CD11b dim myeloid cells from GC patients in the presence or absence of 50% GC patient serum. A proportion of samples were either co-cultured in the presence or absence of IL-6 and/or IL-8 neutralizing abs and/or nor-NOHA, and the proportion of a IFN-γ expressing, b granzyme B expressing and c proliferating CD8 + T cells quantitated by flow cytometry. Data representative of three independent experiments ( n = 3). * p

    Journal: Cell Death & Disease

    Article Title: CD45+CD33lowCD11bdim myeloid-derived suppressor cells suppress CD8+ T cell activity via the IL-6/IL-8-arginase I axis in human gastric cancer

    doi: 10.1038/s41419-018-0803-7

    Figure Lengend Snippet: IL-6/IL-8-arginase I blockade reduces CD45 + CD33 low CD11b dim myeloid cell-mediated CD8 + T cell suppression. CFSE-labeled peripheral blood CD8 + T cells from healthy donors were co-cultured for 5 days with peripheral blood CD45 + CD33 low CD11b dim myeloid cells from GC patients in the presence or absence of 50% GC patient serum. A proportion of samples were either co-cultured in the presence or absence of IL-6 and/or IL-8 neutralizing abs and/or nor-NOHA, and the proportion of a IFN-γ expressing, b granzyme B expressing and c proliferating CD8 + T cells quantitated by flow cytometry. Data representative of three independent experiments ( n = 3). * p

    Article Snippet: Human IL-6 (eBioscience), IL-8 (eBioscience), IFN-γ (Biolegend) and arginase I (Cusabio Biotech) were detected.

    Techniques: Labeling, Cell Culture, Expressing, Flow Cytometry, Cytometry

    Peripheral blood CD45 + CD33 low CD11b dim myeloid cell frequency, and IL-6, IL-8, and arginase I levels correlate with GC tumor progression and decreased patient survival. Peripheral blood ( a ) CD45 + CD33 low CD11b dim myeloid cell percentages ( b ) arginase I concentration, ( c ) IL-6 concentration, and ( d ) IL-8 concentration were measured and categorized by patient TNM stage. Kaplan-Meier plots were calculated for overall patient survival according to ( a ) median peripheral blood CD45 + CD33 low CD11b dim myeloid cell percentage (9.2%) or ( b ) median arginase I concentration (7.52 ng/ml) or ( c ) median IL-6 concentration (22.37 pg/ml) or ( d ) median IL-8 concentration (61.61 pg/ml). * p

    Journal: Cell Death & Disease

    Article Title: CD45+CD33lowCD11bdim myeloid-derived suppressor cells suppress CD8+ T cell activity via the IL-6/IL-8-arginase I axis in human gastric cancer

    doi: 10.1038/s41419-018-0803-7

    Figure Lengend Snippet: Peripheral blood CD45 + CD33 low CD11b dim myeloid cell frequency, and IL-6, IL-8, and arginase I levels correlate with GC tumor progression and decreased patient survival. Peripheral blood ( a ) CD45 + CD33 low CD11b dim myeloid cell percentages ( b ) arginase I concentration, ( c ) IL-6 concentration, and ( d ) IL-8 concentration were measured and categorized by patient TNM stage. Kaplan-Meier plots were calculated for overall patient survival according to ( a ) median peripheral blood CD45 + CD33 low CD11b dim myeloid cell percentage (9.2%) or ( b ) median arginase I concentration (7.52 ng/ml) or ( c ) median IL-6 concentration (22.37 pg/ml) or ( d ) median IL-8 concentration (61.61 pg/ml). * p

    Article Snippet: Human IL-6 (eBioscience), IL-8 (eBioscience), IFN-γ (Biolegend) and arginase I (Cusabio Biotech) were detected.

    Techniques: Concentration Assay

    Peripheral blood CD45 + CD33 low CD11b dim myeloid cells suppress CD8 + T cells through the IL-6/IL-8-arginase I (ARG I) axis in GC patients. Circulating IL-6 and IL-8 induces the secretion of arginase I from CD45 + CD33 low CD11b dim myeloid cells via the PI3K-AKT signaling pathway

    Journal: Cell Death & Disease

    Article Title: CD45+CD33lowCD11bdim myeloid-derived suppressor cells suppress CD8+ T cell activity via the IL-6/IL-8-arginase I axis in human gastric cancer

    doi: 10.1038/s41419-018-0803-7

    Figure Lengend Snippet: Peripheral blood CD45 + CD33 low CD11b dim myeloid cells suppress CD8 + T cells through the IL-6/IL-8-arginase I (ARG I) axis in GC patients. Circulating IL-6 and IL-8 induces the secretion of arginase I from CD45 + CD33 low CD11b dim myeloid cells via the PI3K-AKT signaling pathway

    Article Snippet: Human IL-6 (eBioscience), IL-8 (eBioscience), IFN-γ (Biolegend) and arginase I (Cusabio Biotech) were detected.

    Techniques:

    IL-6 and IL-8 induces CD45 + CD33 low CD11b dim myeloid cell production of arginase I via PI3K-AKT signaling. a Serum IL-6 and IL-8 concentration levels in GC patients compared to healthy donors. b Correlations between the proportions of peripheral blood CD45 + CD33 low CD11b dim myeloid cells or serum arginase I concentration and IL-6 or IL-8 serum concentrations in GC patients. c Arginase I expression and production in CD33 low CD11b dim myeloid cells exposed to recombinant IL-6 and/or IL-8, or 50% GC patient serum in the presence or absence of IL-6 and/or IL-8 neutralizing abs as analyzed by western blot and ELISA, respectively. d Arginase I expression in CD45 + CD33 low CD11b dim myeloid cells exposed to 50% GC patient serum in the presence or absence of FLLL32 (STAT3 phosphorylation inhibitor), Wortmannin (PI3K inhibitor), or SB203580 (MAPK inhibitor; top panel). AKT and p-AKT expression levels in CD45 + CD33 low CD11b dim myeloid cells exposed to IL-6 and/or IL-8 (middle panel), or 50% GC patient serum in the presence or absence of IL-6 and/or IL-8 neutralizing abs (bottom panels) as analyzed by western blot. * p

    Journal: Cell Death & Disease

    Article Title: CD45+CD33lowCD11bdim myeloid-derived suppressor cells suppress CD8+ T cell activity via the IL-6/IL-8-arginase I axis in human gastric cancer

    doi: 10.1038/s41419-018-0803-7

    Figure Lengend Snippet: IL-6 and IL-8 induces CD45 + CD33 low CD11b dim myeloid cell production of arginase I via PI3K-AKT signaling. a Serum IL-6 and IL-8 concentration levels in GC patients compared to healthy donors. b Correlations between the proportions of peripheral blood CD45 + CD33 low CD11b dim myeloid cells or serum arginase I concentration and IL-6 or IL-8 serum concentrations in GC patients. c Arginase I expression and production in CD33 low CD11b dim myeloid cells exposed to recombinant IL-6 and/or IL-8, or 50% GC patient serum in the presence or absence of IL-6 and/or IL-8 neutralizing abs as analyzed by western blot and ELISA, respectively. d Arginase I expression in CD45 + CD33 low CD11b dim myeloid cells exposed to 50% GC patient serum in the presence or absence of FLLL32 (STAT3 phosphorylation inhibitor), Wortmannin (PI3K inhibitor), or SB203580 (MAPK inhibitor; top panel). AKT and p-AKT expression levels in CD45 + CD33 low CD11b dim myeloid cells exposed to IL-6 and/or IL-8 (middle panel), or 50% GC patient serum in the presence or absence of IL-6 and/or IL-8 neutralizing abs (bottom panels) as analyzed by western blot. * p

    Article Snippet: Human IL-6 (eBioscience), IL-8 (eBioscience), IFN-γ (Biolegend) and arginase I (Cusabio Biotech) were detected.

    Techniques: Concentration Assay, Expressing, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay

    Supplementation with  Lactobacillus reuteri  100-23 and  Lactobacillus gasseri  311476 mixture reduces inflammatory cytokines. A–F . Plasma levels of interleukin 4 (IL-4), interleukin 10 (IL-10), interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP-1), interleukin 8 (IL-8), and granulocyte colony-stimulating factor (G-CSF) in control mice (CT), in mice receiving lactobacilli (Lrg), in mice transplanted with BaF3 cells (BaF3) and in mice transplanted with BaF3 cells and receiving lactobacilli (BaF3-Lrg). Data with different superscript letters are significantly different (p

    Journal: PLoS ONE

    Article Title: Restoring Specific Lactobacilli Levels Decreases Inflammation and Muscle Atrophy Markers in an Acute Leukemia Mouse Model

    doi: 10.1371/journal.pone.0037971

    Figure Lengend Snippet: Supplementation with Lactobacillus reuteri 100-23 and Lactobacillus gasseri 311476 mixture reduces inflammatory cytokines. A–F . Plasma levels of interleukin 4 (IL-4), interleukin 10 (IL-10), interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP-1), interleukin 8 (IL-8), and granulocyte colony-stimulating factor (G-CSF) in control mice (CT), in mice receiving lactobacilli (Lrg), in mice transplanted with BaF3 cells (BaF3) and in mice transplanted with BaF3 cells and receiving lactobacilli (BaF3-Lrg). Data with different superscript letters are significantly different (p

    Article Snippet: Plasma interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8) and granulocyte colony-stimulating factor (G-CSF) were measured using a customized multiplex kit (Bio-Rad, Nazareth, Belgium) with the Luminex technology (Bio-Plex, Bio-Rad).

    Techniques: Mouse Assay

    (a) The percentage of blood T cells migrating within three-dimensional collagen gels after 6 days of interleukin-2 (IL-2) stimulation compared with the unstimulated control ( n = 5, P

    Journal: Immunology

    Article Title: Characterization of the migration of lung and blood T cells in response CXCL12 in a three-dimensional matrix

    doi: 10.1111/j.1365-2567.2010.03257.x

    Figure Lengend Snippet: (a) The percentage of blood T cells migrating within three-dimensional collagen gels after 6 days of interleukin-2 (IL-2) stimulation compared with the unstimulated control ( n = 5, P

    Article Snippet: interleukin-2

    Techniques:

    a - f Mitochondrial proteins and inflammatory cytokines levels in explant cultured media in SIM tissue and surrounding matched-normal tissue. Wilcoxon matched-pairs signed rank tests demonstrated significantly increased levels of a cytochrome c (n=12), b SMAC/Diablo (n=8), c IL-1beta (n=12), d IL-6 (n=12), e IL-8 (n=12) and f TNF-alpha (n=12) in SIM tissue compared to surrounding normal epithelium. * p ≤ 0.05, ** p ≤ 0.005 and *** p ≤ 0.0005

    Journal: BMC Cancer

    Article Title: Changes in mitochondrial stability during the progression of the Barrett’s esophagus disease sequence

    doi: 10.1186/s12885-016-2544-2

    Figure Lengend Snippet: a - f Mitochondrial proteins and inflammatory cytokines levels in explant cultured media in SIM tissue and surrounding matched-normal tissue. Wilcoxon matched-pairs signed rank tests demonstrated significantly increased levels of a cytochrome c (n=12), b SMAC/Diablo (n=8), c IL-1beta (n=12), d IL-6 (n=12), e IL-8 (n=12) and f TNF-alpha (n=12) in SIM tissue compared to surrounding normal epithelium. * p ≤ 0.05, ** p ≤ 0.005 and *** p ≤ 0.0005

    Article Snippet: The levels of cytokines interleukin-8 (IL-8), interleukin-6 (IL-6), interleukin-1β (IL-1β) and tumour necrosis factor-α (TNF-α) were measured using a multiplex assay from Mesoscale Discovery® (Gaithersburg, MD, USA) using protocols as per manufactures’ instructions.

    Techniques: Cell Culture

    Th1 responses that are dominant over Th2 responses control L. donovani infection. The frequencies of CD4 + and CD8 + T cells producing IFN-γ and IL-4 in the indicated groups were assessed using single-cell splenocyte suspensions left unstimulated or restimulated overnight with gp63 (5 μg/ml). (A) The frequencies of cells producing IFN-γ were assessed 10 days after vaccination both before and after 3 months of infection. lip, liposome encapsulation; Lip, empty liposomes. (B) The frequencies of cells producing IFN-γ were assessed 12 weeks after vaccination both before and after 3 months of infection. (C) Mice were challenged with L. donovani at 10 days, and the frequencies of cells producing IL-4 were determined at 3 months both before and after infectious challenge. (D) Frequencies of cells producing intracellular IL-4 were also determined in animals at 12 weeks postimmunization both before and after 3 months of infectious challenge. Data represent means ± standard errors of the means of the results obtained with three individual mice.

    Journal: Infection and Immunity

    Article Title: gp63 in Stable Cationic Liposomes Confers Sustained Vaccine Immunity to Susceptible BALB/c Mice Infected with Leishmania donovani ▿

    doi: 10.1128/IAI.00611-07

    Figure Lengend Snippet: Th1 responses that are dominant over Th2 responses control L. donovani infection. The frequencies of CD4 + and CD8 + T cells producing IFN-γ and IL-4 in the indicated groups were assessed using single-cell splenocyte suspensions left unstimulated or restimulated overnight with gp63 (5 μg/ml). (A) The frequencies of cells producing IFN-γ were assessed 10 days after vaccination both before and after 3 months of infection. lip, liposome encapsulation; Lip, empty liposomes. (B) The frequencies of cells producing IFN-γ were assessed 12 weeks after vaccination both before and after 3 months of infection. (C) Mice were challenged with L. donovani at 10 days, and the frequencies of cells producing IL-4 were determined at 3 months both before and after infectious challenge. (D) Frequencies of cells producing intracellular IL-4 were also determined in animals at 12 weeks postimmunization both before and after 3 months of infectious challenge. Data represent means ± standard errors of the means of the results obtained with three individual mice.

    Article Snippet: Horseradish peroxidase-conjugated anti-mouse immunoglobulin G1 (IgG1) and IgG2a, fluorescein isothiocyanate (FITC)-conjugated monoclonal antibodies (MAbs) to mouse CD4 and CD8, phycoerythrin-labeled anti-mouse gamma interferon (IFN-γ) and interleukin-4 (IL-4), and Perm-2 buffer were purchased from BD Biosciences, San Diego, CA.

    Techniques: Infection, Mouse Assay

    Analysis of TLR signaling in Smcr8 −/− and Smcr8 I2T/I2T myeloid cells. ( A ) Peritoneal macrophages harvested from pedigrees R1418 [REF, Smcr8 +/+ ( n = 8); HET, Smcr8 +/p ( n = 15); VAR, Smcr8 p/p ( n = 2)] and R3960 [REF, Smcr8 +/+ ( n = 11); HET, Smcr8 +/p2 ( n = 18); VAR, Smcr8 p2/p2 ( n = 5)] were stimulated with 25 μg/mL of CpG. TNF in the culture medium as measured by ELISA 4 h later. ( B ) Manhattan plot showing P values of association between the phenotype of elevated TNF production in response to CpG and mutations identified in the two pedigrees in A calculated using a recessive model of inheritance. The −log 10 P values are plotted versus the chromosomal positions of mutations. Horizontal red and purple lines represent thresholds of P = 0.05 with or without Bonferroni correction, respectively. The P value for linkage of Smcr8 mutations with the elevated TNF production is indicated. ( C and D ) ELISA analysis of TNF secretion by peritoneal macrophages ( n = 5 or 6 mice per genotype) stimulated for 6 h with indicated concentrations of endosomal TLR ligands poly(I:C), R848, and CpG ( C ) or surface TLR ligands Pam3CSK4 and LPS ( D ). ( E and F ) ELISA analysis of IL-6 secretion by BMDCs ( n = 4 mice per genotype) stimulated for 6 h with indicated concentrations of poly(I:C), R848, and CpG ( E ) or Pam3CSK4 and LPS ( F ). In A and C – F , data points represent individual mice. Data are representative of one ( A and B ) or three independent experiments ( C – F ). Data are expressed as means ± SD, and the significance of differences between genotypes was determined by one-way ANOVA with Dunnett’s multiple comparisons ( C and D ) or unpaired Student’s t test ( E and F ) (* P

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

    Article Title: Excessive endosomal TLR signaling causes inflammatory disease in mice with defective SMCR8-WDR41-C9ORF72 complex function

    doi: 10.1073/pnas.1814753115

    Figure Lengend Snippet: Analysis of TLR signaling in Smcr8 −/− and Smcr8 I2T/I2T myeloid cells. ( A ) Peritoneal macrophages harvested from pedigrees R1418 [REF, Smcr8 +/+ ( n = 8); HET, Smcr8 +/p ( n = 15); VAR, Smcr8 p/p ( n = 2)] and R3960 [REF, Smcr8 +/+ ( n = 11); HET, Smcr8 +/p2 ( n = 18); VAR, Smcr8 p2/p2 ( n = 5)] were stimulated with 25 μg/mL of CpG. TNF in the culture medium as measured by ELISA 4 h later. ( B ) Manhattan plot showing P values of association between the phenotype of elevated TNF production in response to CpG and mutations identified in the two pedigrees in A calculated using a recessive model of inheritance. The −log 10 P values are plotted versus the chromosomal positions of mutations. Horizontal red and purple lines represent thresholds of P = 0.05 with or without Bonferroni correction, respectively. The P value for linkage of Smcr8 mutations with the elevated TNF production is indicated. ( C and D ) ELISA analysis of TNF secretion by peritoneal macrophages ( n = 5 or 6 mice per genotype) stimulated for 6 h with indicated concentrations of endosomal TLR ligands poly(I:C), R848, and CpG ( C ) or surface TLR ligands Pam3CSK4 and LPS ( D ). ( E and F ) ELISA analysis of IL-6 secretion by BMDCs ( n = 4 mice per genotype) stimulated for 6 h with indicated concentrations of poly(I:C), R848, and CpG ( E ) or Pam3CSK4 and LPS ( F ). In A and C – F , data points represent individual mice. Data are representative of one ( A and B ) or three independent experiments ( C – F ). Data are expressed as means ± SD, and the significance of differences between genotypes was determined by one-way ANOVA with Dunnett’s multiple comparisons ( C and D ) or unpaired Student’s t test ( E and F ) (* P

    Article Snippet: The following reagents were used: Pam3CSK4 and R848 (InvivoGen); poly(I:C) (GE Healthcare); LPS and MALP-2 (Enzo Life Sciences); CpG-ODN 1668 (Sigma-Aldrich); and mouse IL-6, IL-12p40, and TNF-α Ready-SET-Go kits (eBioscience).

    Techniques: Enzyme-linked Immunosorbent Assay, Mouse Assay