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: iScience

Article Title: Identification of actionable targets for breast cancer intervention using a diversity outbred mouse model

doi: 10.1016/j.isci.2023.106320

Figure Lengend Snippet: TIEG1 regulates tumor immunity and tumor growth. Tumor growth was monitored using Kaplan-Meier tumor-free survival. p < 0.05 was considered significant. (A) C57BL/6 mice and C57BL/6 HER2 Tg mice with (n = 10) or without TIEG1 (n = 9) were electrovaccinated with pE2TM. (B and C) IFN-γ producing T cells were measured by ELISPOT assay. Data are represented as the average and SD in box and whisker plots. (D) Growth of implanted E0771/E2 tumor in HER2 Tg mice with or without TIEG1. (E) Onset age of spontaneous tumors in NeuT mice with or without TIEG1 (n = 9 and 7, respectively). (F) TIEG1 expression measured by qPCR. Data are represented as pooled averages +/− SEM. (G and H) TIEG1/Klf10 ( top panels ) and Cd3d (bottom panel) expression measured by scRNA sequencing.

Article Snippet: Splenocytes were incubated in ELISPOT plates coated with IFN-γ capture mAb for 48 hrs with recombinant HER2 protein (ecd-Fc fusion; SinoBiological).

Techniques: Enzyme-linked Immunospot, Whisker Assay, Expressing, Sequencing

Journal: Frontiers in Immunology

Article Title: Porous Nanoparticles With Self-Adjuvanting M2e-Fusion Protein and Recombinant Hemagglutinin Provide Strong and Broadly Protective Immunity Against Influenza Virus Infections

doi: 10.3389/fimmu.2018.02060

Figure Lengend Snippet: Enhanced immunogenicity of the combined NPL vaccine vector. (A,B) Survival and weight loss were monitored in influenza virus challenged Balb/c mice following three i.n immunizations with 5 μg (A) or 1 μg (B) of FPM2e or FPM2e:NPL. The percent of surviving mice (left panel) and body weight loss (right panel) following a challenge infection with 4 × LD50 of the mouse adapted X47 virus strain are plotted. (C) Recall responses to M2e-peptide in primed CD4 + T cells following i.n immunizations with 5 or 1 μg of enzymatically active or inactive mutant FPM2e or FPM2e:NPL or empty NPL w/o FPM2e, as indicated. Mean proliferation in isolated splenocytes to M2e peptide is given as mean c.p.m ± S.E.M. (D,E) The production of IFN-γ (D) or IL-17A (E) to recall stimulation with M2e-peptide of the primed CD4 + T cells (as in C ) is given in pg/ml ± SD. (F,G) Representative FACS plots of M2e-tetramer + CD4 + T cells in the lungs of i.n immunized and challenged mice as indicated (F) . The percentage (left panel) and absolute number (right panel) of antigen primed M2e + tetramer CD4 + T cells (G) . (H,I) M2e specific IgG antibodies in serum (H) or IgA antibodies in BAL (I) were measured by ELISA in Balb/c mice immunized i.n. with FPM2e, FPM2e:NPL or PBS (naïve), as indicated, and given as mean log 10 -titers ± SD of 3 independent experiments giving similar results. Statistical significance was calculated by unpaired t -test and p -values are given as * p < 0.05 and ** p < 0.01.

Article Snippet: After washing polyclonal rabbit anti-mouse IFN-γ or anti-IL-17 antibodies (PharMingen, Denmark) at 1 μg/ml in 0.1% BSA/PBS were added to each well and the p-nitrophenyl phosphatase (Sigma Aldrich, Sweden) reaction was visualized using a Titertek Multiscan spectrophotometer (Labsystems, Sweden) at 450 nm.

Techniques: Plasmid Preparation, Infection, Mutagenesis, Isolation, Enzyme-linked Immunosorbent Assay