Journal: International Journal of Inflammation

Article Title: Wogonin Attenuates Ovalbumin Antigen-Induced Neutrophilic Airway Inflammation by Inhibiting Th17 Differentiation

doi: 10.1155/2014/571508

Figure Lengend Snippet: Screening of Kampo extracts and identification of major components that exhibit the inhibitory effects of IL-17 secretion. HJG, HST, OGT, SST, HET, JTT, DKT, or SRT were added at a concentration of 10 μ g/mL to the two-way MLR of splenic lymphocytes obtained from BALB/c and SJL mice and then were incubated for seven days (a). The supernatants were analyzed using ELISA to detect the typical cytokine secretions of IFN- γ ( center ), IL-5 ( right ), and IL-17 ( left ) for Th1, Th2, and Th17, respectively. Increasing concentrations of wogonin or berberine (0.1, 0.3, and 1 μ M) were mixed with a culture of lymphocytes obtained from BALB/c and SJL mice and then incubated for seven days (b). The supernatants were analyzed using ELISA to detect the typical cytokines of IFN- γ (center), IL-4 (right), and IL-17 (left) for Th1, Th2, and Th17, respectively.

Article Snippet: The culture supernatants were assayed for IFN- γ , IL-4, IL-5, and IL-17 using ELISA kits (R&D systems, U.S.A.).

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

Journal: International Journal of Inflammation

Article Title: Wogonin Attenuates Ovalbumin Antigen-Induced Neutrophilic Airway Inflammation by Inhibiting Th17 Differentiation

doi: 10.1155/2014/571508

Figure Lengend Snippet: Suppression of IL-17 by wogonin treatment in the human MLR. For the human two-way MLR (a), wogonin (1 μ M) was exposed to a mixture of human allogeneic lymphocytes and then incubated for seven days. Seven days after incubation, the supernatants were analyzed using ELISA to detect the typical cytokines of IFN- γ (center), IL-4 (right), and IL-17 (left) for Th1, Th2, and Th17, respectively. For the human one-way MLR (b), wogonin (1 μ M) was incubated with human Mo-DCs for two days. Then, the wogonin-treated human Mo-DCs were incubated with human allogeneic naïve CD4 + T cells in the absence of wogonin for seven days. The cells were restimulated with anti-human CD3 and CD28 antibodies for 24 h, and the supernatants were collected for ELISA to detect the typical cytokines of IFN- γ (center), IL-4 (right), and IL-17 (left) for Th1, Th2, and Th17, respectively.

Article Snippet: The culture supernatants were assayed for IFN- γ , IL-4, IL-5, and IL-17 using ELISA kits (R&D systems, U.S.A.).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Inflammation

Article Title: Wogonin Attenuates Ovalbumin Antigen-Induced Neutrophilic Airway Inflammation by Inhibiting Th17 Differentiation

doi: 10.1155/2014/571508

Figure Lengend Snippet: Effects of wogonin on a human Th17 clone. The human alloreactive Th17 clone was established earlier . Following incubation of Th17 with irradiated allogeneic PBMCs in the presence or absence of wogonin (1 μ M) for 24 h, the supernatants were analyzed using ELISA to detect the secretion of IL-17 ((a), left). To assess the proliferation of Th17, the cells were incubated with irradiated allogeneic PBMCs in the presence or absence of wogonin (1 μ M) for 72 h and analyzed using [ 3 H] TdR incorporation during the final 16 h period. The incorporated radioactivity was analyzed using liquid scintillation counting after harvesting ((a), right). Wogonin belongs to flavonoid species (b).

Article Snippet: The culture supernatants were assayed for IFN- γ , IL-4, IL-5, and IL-17 using ELISA kits (R&D systems, U.S.A.).

Techniques: Incubation, Irradiation, Enzyme-linked Immunosorbent Assay, Radioactivity

Journal: PLoS ONE

Article Title: Laminin and Fibronectin Treatment Leads to Generation of Dendritic Cells with Superior Endocytic Capacity

doi: 10.1371/journal.pone.0010123

Figure Lengend Snippet: A , IFN-gamma production by memory T-cells after 24 and 48 h of co-culture with ECM-treated DC pre-loaded with CMV protein. B , T-cell proliferation assays using memory T-cells as responder cells. The proliferation index (PI) was calculated as described in the . C , A representative histogram showing the uptake of fluorescein-labeled CMV protein by DC before and after treatment with ECM. Cells without CMV (filled histogram), cells at 4°C with CMV (solid gray line), control DC with CMV (solid black line), FN treated DC with CMV (dashed black line) and LMN treated DC with CMV (dashed gray line). D , CMV uptake by DC under different culture conditions. Data from is represented as a histogram with n = 3. E , Analysis of conjugate formation between DC and T cells. PKH26-labeled DC were incubated with CFSE-labeled T cells for 45 min (ratio 1DC/2T cell). Conjugate formation was analyzed by flow cytometry by determining the CFSE+ cells amongst the DC population. To rule out phagocytosis or dye diffusion as a cause of double positive cells, conjugate formation was investigated on cells treated with 5 mM EDTA and vortexed for 2 min after incubation. Results depict data from one of three independent experiments. *P<0.05, ** P<0.001.

Article Snippet: IFN-gamma was quantified using an ELISA kit (R&D Systems, Oxford, UK) according to the manufacturer's instructions.

Techniques: Co-Culture Assay, Labeling, Control, Incubation, Flow Cytometry, Diffusion-based Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Atorvastatin (Lipitor) attenuates the effects of aspirin on pancreatic cancerogenesis and the chemotherapeutic efficacy of gemcitabine on pancreatic cancer by promoting M2 polarized tumor associated macrophages

doi: 10.1186/s13046-016-0304-4

Figure Lengend Snippet: Gemcitabine induced systemic and intratumoral Th2 biased cytokine environment, aspirin weakened these effects of gemcitabine, however Lipitor augmented these effects. a . The cytokines in spleen were measured by real time RT-PCR. The results indicated that gemcitabine reduced the levels of IL-6, TNF-α and IL-2, however it increaed the levels of IL-4. Lipitor increased the levels of IL-4 and TGF-β. Aspirin lowered down IL-6. And the results were confirmed by the elisa tests ( d ). b . The cytokines in tumor tissues were measured by real time RT-PCR. The results indicated that gemcitabine reduced the intratumoral levels of IL-6, INF-γ, TNF-α and IL-2, however it elevated the levels of IL-4, IL-10 and TGF-β. Lipitor increased the levels of IL-4, IL-10 and TGF-β. Aspirin lowered down IL-4, IL-10, TGF-β and IL-6. And the results were confirmed by the elisa tests ( e ). c . The arginase in spleens and tumor tissues were detected by real time RT-PCR. The results confirmed the results of FCM, that is, gemcitabine inhibited MDSC, aspirin inhibited MDSC as well, however Lipitor did not affect MDSC significantly. Each experiment was performed in triplicats and the representative one was shown. . ANOVA analysis, SNK-q test, * P < 0.05, ** P < 0.01

Article Snippet: All of the commercial sandwich ELISA kits used for the quantitation of IL-2, IL-4, IL-6, IL-10, IL-12, IFN-γ, TNF-α and TGF-β were purchased from R&D systems Inc..

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

Journal: Journal of Translational Medicine

Article Title: Overexpression of IL-17RC associated with ocular sarcoidosis

doi: 10.1186/1479-5876-12-152

Figure Lengend Snippet: Induction of IL-17RC on CD8 + T cells. (A) Serum levels of IL-17 in 18 healthy controls and 13 patients with ocular sarcoidosis. (B) The frequency of IL-17RC + in cultured CD8 + T cells treated with (IL-17) or without (Con) IL-17 from 4 healthy controls and 4 sarcoidosis patients. *P < 0.05.

Article Snippet: Sera from sarcoidosis patients and healthy controls were centrifuged at 1400 rpm for 15 min after 30 min of clotting and were stored at -80°C, followed by detection of IL-17A using the Human IL-17 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN) according to the manufacture’s protocol.

Techniques: Cell Culture

Journal: Clinical and Experimental Medicine

Article Title: IL-21- and CXCL9-engineered GPC3-specific CAR-T cells combined with PD-1 blockade enhance cytotoxic activities against hepatocellular carcinoma

doi: 10.1007/s10238-024-01473-2

Figure Lengend Snippet: Cytotoxicity assays in vitro. a GPC3 expression of HCC cell lines were analyzed by FCM. Isotype Ab served as a control. b GPC3 expression confirmed by Western blotting analysis. c The secretion of IL-21 or CXCL9 of indicated CAR-T cells determined by ELISA. d Cytokines (IL-2, IFN-γ and TNF-a) released by different CAR-T cells after 24 h coculture with Hep3B cells at 1:1 ratio was measured by ELISA. e Cytotoxicity of indicated CAR-T cells incubated with HepG2, Hep3B, Huh-7 and SK-Hep-1 at 5:1, 10:1, and 20:1 ratio for 24 h. Each data set reflected mean ± SD of triplicates, ns represent p > 0.05, ** p < 0.01 *** p < 0.001. The ELISA results were compared with one-way ANOVA, and the cytotoxicity assay was conducted by two-way ANOVA

Article Snippet: The ELISA kits for IL-2 and IFN-γ were purchased from Sino Biological (Beijing, China), and the ELISA kits for IL-21 and CXCL9 were purchased from Novus Biologicals (Littleton.

Techniques: In Vitro, Expressing, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Incubation, Cytotoxicity Assay