Journal: bioRxiv

Article Title: Modified E2 glycoprotein of hepatitis C virus enhances pro-inflammatory cytokines and protective immune response in mice

doi: 10.1101/2022.02.07.479452

Figure Lengend Snippet: Human monocyte derived macrophage cell line THP-1 was treated with 2.5 µg/ml of modified E2 for 24 h. Secretory cytokines IL-10 and IL-12 in the culture supernatant were quantified by ELISA (panels A and B). The significance level is expressed as *=p<0.05 **= p < 0.005.

Article Snippet: Secretory cytokines present in the cell culture supernatant of macrophages exposed to HCV sE2, sE2 L427Y, or sE2 F442NYT after 24 h were quantified using commercially available paired antibodies for IL-10, IL-12, IL-6, IFN-γ and TNF-α cytokine kit (Sino Biological).

Techniques: Derivative Assay, Modification, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Modified E2 glycoprotein of hepatitis C virus enhances pro-inflammatory cytokines and protective immune response in mice

doi: 10.1101/2022.02.07.479452

Figure Lengend Snippet: Monocyte derived macrophages from healthy donors (n=6) were incubated with 2.5 µg/ml HCV sE2 or modified proteins for 24 h, and secretory cytokines (IL-10, IL-4, IL-12, IFN-γ, IL-6, and TNF-α) in the culture supernatant were quantified by ELISA (panels A-F). The significance level is expressed as *=p<0.05, ** = p < 0.005, *** = p < 0.001, or ns (not significant).

Article Snippet: Secretory cytokines present in the cell culture supernatant of macrophages exposed to HCV sE2, sE2 L427Y, or sE2 F442NYT after 24 h were quantified using commercially available paired antibodies for IL-10, IL-12, IL-6, IFN-γ and TNF-α cytokine kit (Sino Biological).

Techniques: Derivative Assay, Incubation, Modification, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Modified E2 glycoprotein of hepatitis C virus enhances pro-inflammatory cytokines and protective immune response in mice

doi: 10.1101/2022.02.07.479452

Figure Lengend Snippet: IL-4, IL-10, IL-12 and IFN-γ expression (panels A-D) were quantified from sera of unimmunized control, E1/sE2, or E1/sE2 F442NYT vaccinated mice by ELISA. The significance level is expressed as *=p<0.05 ** = p < 0.005, or ns (not significant).

Article Snippet: Secretory cytokines present in the cell culture supernatant of macrophages exposed to HCV sE2, sE2 L427Y, or sE2 F442NYT after 24 h were quantified using commercially available paired antibodies for IL-10, IL-12, IL-6, IFN-γ and TNF-α cytokine kit (Sino Biological).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: CRAC Channel Controls the Differentiation of Pathogenic B Cells in Lupus Nephritis

doi: 10.3389/fimmu.2021.779560

Figure Lengend Snippet: CRAC channel inhibition suppressed B cell differentiation. (A) Blimp-1 expression in B cells from patients with lupus nephritis (LN) or healthy controls (HC) was measured by Western blot and representative bands of six independent samples. (B, C) Naive B cells from LN or HC were cultured in the presence of anti-IgM (5 μg/ml), anti-CD40 (1 μg/ml), IL-2 (20 ng/ml), IL-4 (20 ng/ml), and IL-21 (50 ng/ml) for 8 days. Percentages of CD19 low CD138 + plasma cells were determined by flow cytometry. Representative counter plots are shown and data from five samples. Data are mean ± SEM. *p < 0.05 by t-test. (D) GSEA plot for the gene set of B-cell differentiation showing the enrichment scores for YM-58483 or vehicle-treated B cells (n = 3). (E) Blimp-1 expression in B cells treated with YM58483 or vehicle was measured by Western blot and representative bands of six independent samples. (F) Blimp-1, ORAI1 expression in B cells treated with ORAI1 siRNA or scramble siRNA was measured by Western blot. Representative bands of six samples are shown. (G, H) CD138 and IgG expression in B cells treated with YM-58483 or vehicle. Representative counter plots are shown, and the percentages of CD138 + or CD138 + IgG + cells are summarized in dot plot with bar plot (n = 5). (I, J) CD138 and IgG expression in B cells treated with ORAI1 siRNA or scramble siRNA. Representative counter plots are shown, and the percentage of CD138 + or CD138 + IgG + cells are summarized in dot plot with bar plot (n = 5). For the Western blot data, relative expression values to GAPDH are indicated above each lane. *p < 0.05, **p < 0.01 by paired t-test.

Article Snippet: Cells were stimulated with antihuman IgM (5 μg/ml, Sigma, #10759) and antihuman CD40 (1 μg/ml, Bioxcell, #BE0189) antibodies in the presence of interleukin (IL)-2 (20 ng/ml, PeproTech), IL-4 (25 ng/ml, Sino Biological) and IL-21 (50 ng/ml, Sino Biological) for 8 days.

Techniques: Inhibition, Cell Differentiation, Expressing, Western Blot, Cell Culture, Flow Cytometry

Journal: Theranostics

Article Title: Grafted human ESC-derived astroglia repair spinal cord injury via activation of host anti-inflammatory microglia in the lesion area

doi: 10.7150/thno.70929

Figure Lengend Snippet: Grafted astroglia activate anti-inflammatory microglia via IL-4 signaling. (A-C) Representative images showing CD68 (red) and IL-4R (green) staining in the lesion center at 4 weeks post-transplantation. A1-C1 are higher magnification images of boxed areas in A-C ; A1'-C1' are three-dimensional models of the microglia in A1-C1 . Scale bars: 30 μm (A-C) , 5 μm (A1-C1, A1'-C1') . (D) Quantification of IL-4R + CD68 + cells relative to all CD68 + cells ( n = 5 per group). Brown-Forsythe and Welch ANOVA followed by Dunnett's T3 multiple comparisons test. Data are mean ± SD; ** p < 0.01, *** p < 0.001. (E) Western blotting for IL-4, p-STAT6, and Arg1 expression in the lesion area at 3 to 28 days after transplantation. (F-H) Quantification of IL-4, p-STAT6 and Arg1 expression at 3 to 28 days ( n = 3 per group). One-way ANOVA followed by Tukey's multiple comparisons test for each time point; once one-way ANOVA failed, Brown-Forsythe and Welch ANOVA followed by Dunnett's T3 multiple comparisons test or unpaired t test with Welch's correction. Data are mean ± SD; * p < 0.05, ** p < 0.01, n.s., not significant. (I) Diagram depicting the model for anti-inflammatory polarization of microglia to promote repair of the spinal cord lesion after astroglial transplantation.

Article Snippet: Primary antibodies used for WB included: monoclonal anti-PDGFR-β (Abcam cat. #ab32570; RRID: AB_2262874; 1:5000), monoclonal anti-fibronectin (Abcam cat. #ab2413; RRID: AB_777165; 1:5000), monoclonal anti-IL-4 (Sino Biological cat. #11846-R404; 1:1000), monoclonal anti-phospho-STAT6 (CST cat. #56554; RRID: AB_2799514; 1:500), polyclonal anti-Arg1 (GeneTex cat. #GTX109242; RRID: AB_2036264; 1:1000), polyclonal anti-Iba1 (Abcam cat. #ab5076; RRID: AB_2224402; 1:1000).

Techniques: Staining, Transplantation Assay, Western Blot, Expressing

Journal: bioRxiv

Article Title: A Photonic Biosensor-Integrated Tissue Chip Platform for Real-Time Sensing of Lung Epithelial Inflammatory Markers

doi: 10.1101/2022.06.21.497028

Figure Lengend Snippet: a) Ring functionalization scheme. Each control antibody is matched to the antibody isotype of the corresponding capture antibody. “High” and “Low” for the mouse antibodies refers to the concentration used to match that of IL-1β vs IL-8, since their stock concentrations were different. b) Nonspecific binding for two IL-1β channels (both represent control-subtracted relative shifts) over about 3 hours, and c) relative shifts after adding analyte. d) a significant relative blueshift is seen for IL-6, meaning the isotype control antibody used is not an ideal match. e) Significant redshifts seen for IL-6 once analyte is added. f) Nonspecific and g) analyte-specific shifts for IL-8.

Article Snippet: The bottom rings were spotted with antibodies to IL-1β (R&D Systems) at 500 μg/mL, IL-6 (Biolegend, San Diego, CA) at 650 μg/mL, IL-8 (Sino Biological, Wayne, PA) at 1000 μg/mL, and C-reactive protein (CRP; Ortho-Clinical Diagnostics, Rochester, NY) at 990 μg/mL ( ).

Techniques: Concentration Assay, Binding Assay

Journal: bioRxiv

Article Title: A Photonic Biosensor-Integrated Tissue Chip Platform for Real-Time Sensing of Lung Epithelial Inflammatory Markers

doi: 10.1101/2022.06.21.497028

Figure Lengend Snippet: Photonic sensor calibration for various analytes: a) CRP (n = 6 channels), b) IL-1β (n = 3), c) IL-6 (n = 3) and d) IL-8 (n = 4). All responses were fit with four-parameter logistic curves. Error bars represent standard error.

Article Snippet: The bottom rings were spotted with antibodies to IL-1β (R&D Systems) at 500 μg/mL, IL-6 (Biolegend, San Diego, CA) at 650 μg/mL, IL-8 (Sino Biological, Wayne, PA) at 1000 μg/mL, and C-reactive protein (CRP; Ortho-Clinical Diagnostics, Rochester, NY) at 990 μg/mL ( ).

Techniques:

Journal: Food & Nutrition Research

Article Title: Protective effect of free phenolics from Lycopus lucidus Turcz. root on carbon tetrachloride-induced liver injury in vivo and in vitro

doi: 10.29219/fnr.v62.1398

Figure Lengend Snippet: Effects of FPLR (100 mg/kg bw) on the levels of TNF-α, IL-6, IL-8, COX-2, iNOS, and Caspase-3 with the immunohistochemical method (40×). Statistical analysis was based on the values of integrated optical density. Values expressed as mean ± SD in each group ( n = 10). The different capital letters indicate significant differences among the different groups at p < 0.01.

Article Snippet: Rabbit anti-tumor necrosis factor-α (TNF-α), rabbit anti-interleukin-6 (IL-6), rabbit anti-interleukin-8 (IL-8), rabbit anti-cyclooxygenase-2 (COX-2), rabbit anti-caspase-3, rabbit anti-inducible nitric oxide synthase (iNOS) antibodies, and 3,3’-diaminobenzidine tetrahydrochloride (DAB) chromogenic agent were purchased from Servicebio Chemical Co. (Wuhan, China).

Techniques: Immunohistochemical staining