dy3144 05  (R&D Systems)

Journal: JOR Spine

Article Title: Macrophage Subtypes Distinctly Impact Intervertebral Disc Inflammation and Extracellular Matrix Composition

doi: 10.1002/jsp2.70143

Figure Lengend Snippet: (A) Daily and (B) cumulative NO release from IVD explants with and without TNFα in macrophage media ( n = 3). Cumulative (C) TNFα, (D) IL‐6, (E) IL‐13, and (F) CCL2 in media supernatants of IVD explants with and without TNFα stimulation ( n = 3; Black * p < 0.05 IVD significantly different from IVD + TNFα within timepoint, colored * p < 0.05 Group of corresponding color significantly different between timepoints).

Article Snippet: Production of inflammatory cytokines and chemokines was also evaluated using the Rat CCL2 DuoSet ELISA (R&D Systems, Cat#. DY3144‐05) and the LEGENDPlex Rat Th Cytokine Panel V02 (BioLegend, Cat#. 741220) multiplex assay containing 13 analytes: TNFα, IL‐6, IL‐13, IFNγ, IL‐17A, IL‐17F, IL‐5, IL‐2, IL‐4, IL‐10, IL‐22, GM‐CSF, and IL‐9.

Techniques:

Journal: JOR Spine

Article Title: Macrophage Subtypes Distinctly Impact Intervertebral Disc Inflammation and Extracellular Matrix Composition

doi: 10.1002/jsp2.70143

Figure Lengend Snippet: (A) Daily and (B) cumulative NO release from TNFαα‐stimulated IVD explants alone and co‐cultured with M1 macrophages (IVD + TNFα: n = 3, IVD + TNFα + M1: n = 6). Cumulative concentrations of (C) TNFα, (D) IL‐6, (E) IL‐13, and (F) CCL2 in media supernatants from M1 macrophage‐IVD explant co‐culture in TNFα ( n = 3). (G) Daily and (H) cumulative NO release from TNFα‐stimulated IVD explants alone and co‐cultured with M2 macrophages (IVD + TNFα: n = 3, IVD + TNFα + M2: n = 6). Cumulative concentrations of (I) TNFα, (J) IL‐6, (K) IL‐13, and (L) CCL2 in media supernatants from M2 macrophage‐IVD explant co‐culture in TNFα ( n = 3; Black * p < 0.05 IVD + TNFα significantly different from IVD + TNFα + M1/M2 within timepoint, colored * p < 0.05 Group of corresponding color significantly different between timepoints, ^ p < 0.05 IVD + TNFα + M1/M2 signficantly different from TNFα + M1/M2, + p < 0.05 IVD + TNFα significantly different from TNFα + M1/M2).

Article Snippet: Production of inflammatory cytokines and chemokines was also evaluated using the Rat CCL2 DuoSet ELISA (R&D Systems, Cat#. DY3144‐05) and the LEGENDPlex Rat Th Cytokine Panel V02 (BioLegend, Cat#. 741220) multiplex assay containing 13 analytes: TNFα, IL‐6, IL‐13, IFNγ, IL‐17A, IL‐17F, IL‐5, IL‐2, IL‐4, IL‐10, IL‐22, GM‐CSF, and IL‐9.

Techniques: Cell Culture, Co-Culture Assay

Journal: Veterinary sciences

Article Title: IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine Keratinocytes.

doi: 10.3390/vetsci12010055

Figure Lengend Snippet: Figure 1. The effects of Th1 or Th2 cytokine stimulation on the production of IL-6, CXCL8, and CCL2 from canine progenitor epidermal keratinocytes (CPEKs). (A) CPEKs stimulated with an IFN-γ/TNF- α mixture showed increased IL-6 concentrations compared to vehicle (** p < 0.01, unpaired t-test). (B) Supernatants from CPEKs stimulated with TNF-α (** p < 0.01, Mann–Whitney U test) and the mixture of IFN-γ/TNF-α (** p < 0.01, unpaired t-test) showed significantly increased CXCL8 concen- trations. CXCL8 levels were more elevated in cells stimulated by TNF-α than in those stimulated by the mixture of IFN-γ/TNF-α (+ p < 0.01, Mann–Whitney U test). The supernatants from CPEKs incubated with IFN-γ showed decreased CXCL8 concentrations (** p < 0.01, unpaired t-test). (C) The supernatants from CPEKs incubated with IFN-γ, TNF-α, and the IFN-γ/TNF-α mixture exhibited significantly increased CCL2 concentrations compared to vehicle (** p < 0.01, Mann–Whitney U test). Among them, those from CPEKs incubated with the IFN-γ/TNF-α mixture showed significantly increased CCL2 concentrations compared to those incubated with either IFN-γ or TNF-α (+ p < 0.01 TNF-α versus the IFN-γ/TNF-α mixture, ¶ p < 0.05, IFN-γ versus the IFN-γ/TNF-α mixture, Mann– Whitney U test). In addition, the supernatants of CPEKs incubated with IFN-γ showed significantly increased CCL2 concentrations compared to those from CPEKs incubated with TNF-α (# p < 0.01, unpaired t-test). Data represent the mean of five independent experiments ± standard deviation.

Article Snippet: Canine IL-6, IL-10, IL-1ß, IL-12, CXCL8, and CCL2 DuoSet ELISA kits (R&D systems) were used according to the manufacturer’s instructions.

Techniques: MANN-WHITNEY, Incubation, Standard Deviation

Journal: Veterinary sciences

Article Title: IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine Keratinocytes.

doi: 10.3390/vetsci12010055

Figure Lengend Snippet: Figure 2. Dose-dependent effects on the production of IL-6, CXCL8, and CCL2 from canine progenitor epidermal keratinocytes (CPEKs). (A) CPEKs incubated with 60 and 90 ng/mL of the IFN-γ/TNF-α mixture showed significantly increased IL-6 concentrations in their culture medium compared to those treated with 10 ng/mL of the IFN-γ/TNF-α mixture (Kruskal–Wallis test with Dunnett’s test). (B) No significant differences in CXCL8 concentration were detected in response to treatment with different concentrations of TNF-α (10, 30, 60, and 90 ng/mL). (C) No significant differences in CXCL8 concentration were detected in response to treatment with different doses of the mixture of IFN- γ/TNF-α (10, 30, 60, or 90 ng/mL). (D) CPEKs incubated with 30, 60, or 90 ng/mL of IFN-γ showed significantly increased CCL2 concentrations in their culture medium compared to those treated with 10 ng/mL of IFN-γ (ANOVA with Tukey’s HSD). (E) CPEKs incubated with 90 ng/mL of TNF-α also showed significantly increased CCL2 concentrations in their culture medium than those treated with 10 ng/mL of TNF-α (ANOVA with Tukey’s HSD). (F) CPEKs incubated with 30, 60, or 90 ng/mL of the IFN-γ/TNF-α mixture showed significantly increased CCL2 concentrations in their culture medium than those treated with 10 ng/mL of the same mixture (Kruskal–Wallis test with Dunnett’s test). Data represent the mean of five independent experiments ± standard deviation. * p < 0.05, ** p < 0.01.

Article Snippet: Canine IL-6, IL-10, IL-1ß, IL-12, CXCL8, and CCL2 DuoSet ELISA kits (R&D systems) were used according to the manufacturer’s instructions.

Techniques: Incubation, Concentration Assay, Standard Deviation