Journal: Journal of Innate Immunity

Article Title: Deletion of Irf3 and Irf7 Genes in Mice Results in Altered Interferon Pathway Activation and Granulocyte-Dominated Inflammatory Responses to Influenza A Infection

doi: 10.1159/000450705

Figure Lengend Snippet: Quantification of IFN type I and type III protein levels in KO and WT mice infected with PR8M IAV. After intranasal infection of female mice at the age of 8–11 weeks (n = 5–8) with PBS or 2 × 105 FFU PR8M, BAL was collected at the indicated days and used for IFNA (a), IFNB1 (b) or IFNL2/3 (IL-28A/B; c) detection by ELISA. Each sample was measured in technical duplicates. Values from individual mice, the mean and SEM are presented. The blue line indicates the detection limit of each specific assay. Protein measurements that were below the detection limit were set to the detection limit. The significance of differences in protein concentrations between mutant mice and WT controls were calculated at each time point by the two-tailed Mann-Whitney U test. n.s. = No significance. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Supernatants were aliquoted, stored at −70°C and used to detect the IFNA, IFNB1, and IFNL2/3 (also referred to as IL-28A/B) protein concentration by the respective ELISA kits from PBL Assay Science (Piscataway, N.J., USA).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Mutagenesis, Two Tailed Test, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Type I and Type III Interferons Display Different Dependency on Mitogen-Activated Protein Kinases to Mount an Antiviral State in the Human Gut

doi: 10.3389/fimmu.2017.00459

Figure Lengend Snippet: Induction of immune response in human mini-gut organoids after mammalian reovirus (MRV) infection . Organoids were infected with MRV (multiplicity of infection = 0.5), quantitative real-time (qRT)-PCR and ELISA were used to detect (A) a time course of transcriptional upregulation of both type I interferons (IFNs) (β) and type III IFN (λ2/3) IFNs (B) 24 hpi the production and secretion of IFN proteins in the supernatant of infected organoids and (C) a time course of transcriptional upregulation of the IFN-stimulated genes Viperin and IFIT1. qRT-PCR data were normalized to TBP and HPRT1 (housekeeping genes) and are expressed relative to uninfected organoids at each time point. qRT-PCR data and ELISA data represent the mean values of three independent experiments. Error bars indicate the SD. The blue and red lines in (B) demarcate the limit of detection of our ELISA for type I and type III IFNs, respectively.

Article Snippet: IFNβ and IFNL2/3 contained in the supernatant of cells were quantified using the human IFN-beta ELISA kit and DIY IFNLR 2/3 ELISA kit both from PBL-Interferon Source, per manufacturer’s instructions.

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

Journal: Frontiers in Immunology

Article Title: Type I and Type III Interferons Display Different Dependency on Mitogen-Activated Protein Kinases to Mount an Antiviral State in the Human Gut

doi: 10.3389/fimmu.2017.00459

Figure Lengend Snippet: Expression pattern of interferon (IFN) mRNA and protein in human intestinal epithelial cells upon viral infection (A) . Relative quantification of type I IFN (β) and type III IFN (λ2/3) transcripts during the course of mammalian reovirus (MRV) (multiplicity of infection = 1) infection of T84 cells. Data are normalized to TBP and HPRT1 and are expressed relative to uninfected cells at each time point. (B) Quantification of type (IFNβ) and type III (IFN λ2/3) protein levels by ELISA in supernatants of uninfected or MRV-infected T84 cells. The blue and red dashed lines demarcate the limit of detection of our ELISA for type I and type III IFNs, respectively. n.d., not detectable. Data represent the mean values of three independent experiments. Error bars indicate the SD. **** P < 0.0001 (unpaired t -test).

Article Snippet: IFNβ and IFNL2/3 contained in the supernatant of cells were quantified using the human IFN-beta ELISA kit and DIY IFNLR 2/3 ELISA kit both from PBL-Interferon Source, per manufacturer’s instructions.

Techniques: Expressing, Infection, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Type I and Type III Interferons Display Different Dependency on Mitogen-Activated Protein Kinases to Mount an Antiviral State in the Human Gut

doi: 10.3389/fimmu.2017.00459

Figure Lengend Snippet: Induction of immune response in human mini-gut organoids after mammalian reovirus (MRV) infection . Organoids were infected with MRV (multiplicity of infection = 0.5), quantitative real-time (qRT)-PCR and ELISA were used to detect (A) a time course of transcriptional upregulation of both type I interferons (IFNs) (β) and type III IFN (λ2/3) IFNs (B) 24 hpi the production and secretion of IFN proteins in the supernatant of infected organoids and (C) a time course of transcriptional upregulation of the IFN-stimulated genes Viperin and IFIT1. qRT-PCR data were normalized to TBP and HPRT1 (housekeeping genes) and are expressed relative to uninfected organoids at each time point. qRT-PCR data and ELISA data represent the mean values of three independent experiments. Error bars indicate the SD. The blue and red lines in (B) demarcate the limit of detection of our ELISA for type I and type III IFNs, respectively.

Article Snippet: IFNβ and IFNL2/3 contained in the supernatant of cells were quantified using the human IFN-beta ELISA kit and DIY IFNLR 2/3 ELISA kit both from PBL-Interferon Source, per manufacturer’s instructions.

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

Journal: Frontiers in Immunology

Article Title: Type I and Type III Interferons Display Different Dependency on Mitogen-Activated Protein Kinases to Mount an Antiviral State in the Human Gut

doi: 10.3389/fimmu.2017.00459

Figure Lengend Snippet: Expression pattern of interferon (IFN) mRNA and protein in human intestinal epithelial cells upon viral infection (A) . Relative quantification of type I IFN (β) and type III IFN (λ2/3) transcripts during the course of mammalian reovirus (MRV) (multiplicity of infection = 1) infection of T84 cells. Data are normalized to TBP and HPRT1 and are expressed relative to uninfected cells at each time point. (B) Quantification of type (IFNβ) and type III (IFN λ2/3) protein levels by ELISA in supernatants of uninfected or MRV-infected T84 cells. The blue and red dashed lines demarcate the limit of detection of our ELISA for type I and type III IFNs, respectively. n.d., not detectable. Data represent the mean values of three independent experiments. Error bars indicate the SD. **** P < 0.0001 (unpaired t -test).

Article Snippet: IFNβ and IFNL2/3 contained in the supernatant of cells were quantified using the human IFN-beta ELISA kit and DIY IFNLR 2/3 ELISA kit both from PBL-Interferon Source, per manufacturer’s instructions.

Techniques: Expressing, Infection, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay