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  1. Article ; Online: Luciferase reporter assays to monitor interferon signaling modulation by SARS-CoV-2 proteins

    Maximilian Hirschenberger / Manuel Hayn / Alexandre Laliberté / Lennart Koepke / Frank Kirchhoff / Konstantin Maria Johannes Sparrer

    STAR Protocols, Vol 2, Iss 4, Pp 100781- (2021)

    2021  

    Abstract: Summary: We present a protocol for analyzing the impact of SARS-CoV-2 proteins in interferon signaling using luciferase reporter assays. Here, the induction of defined promoters can be quantitatively assessed with high sensitivity and broad linear range. ...

    Abstract Summary: We present a protocol for analyzing the impact of SARS-CoV-2 proteins in interferon signaling using luciferase reporter assays. Here, the induction of defined promoters can be quantitatively assessed with high sensitivity and broad linear range. The results are similar to those obtained using qPCR to measure endogenous mRNA induction. The assay requires stringent normalization and confirmation of the results in more physiological settings. The protocol is adaptable for other viruses and other innate immune stimuli.For complete details on the use and execution of this protocol, please refer to Hayn et al. (2021).
    Keywords Cell-based assays ; Immunology ; Microbiology ; Molecular biology ; Gene expression ; Molecular/chemical probes ; Science (General) ; Q1-390
    Language English
    Publishing date 2021-12-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: Vpr attenuates antiviral immune responses and is critical for full pathogenicity of SIVmac239 in rhesus macaques

    Alexandre Laliberté / Caterina Prelli Bozzo / Christiane Stahl-Hennig / Victoria Hunszinger / Simone Joas / Ulrike Sauermann / Berit Roshani / Antonina Klippert / Maria Daskalaki / Kerstin Mätz-Rensing / Nicole Stolte-Leeb / Gregory K. Tharp / Dietmar Fuchs / Prachi Mehrotra Gupta / Guido Silvestri / Sydney A. Nelson / Laura Parodi / Luis Giavedoni / Steven E. Bosinger /
    Konstantin M.J. Sparrer / Frank Kirchhoff

    iScience, Vol 26, Iss 12, Pp 108351- (2023)

    2023  

    Abstract: Summary: The accessory viral protein R (Vpr) is encoded by all primate lentiviruses. Vpr counteracts DNA repair pathways, modulates viral immune sensing, and induces cell-cycle arrest in cell culture. However, its impact in vivo is controversial. Here, ... ...

    Abstract Summary: The accessory viral protein R (Vpr) is encoded by all primate lentiviruses. Vpr counteracts DNA repair pathways, modulates viral immune sensing, and induces cell-cycle arrest in cell culture. However, its impact in vivo is controversial. Here, we show that deletion of vpr is associated with delayed viral replication kinetics, rapid innate immune activation, development and maintenance of strong B and T cell responses, and increased neutralizing activity against SIVmac239 in rhesus macaques. All wild-type SIVmac239-infected animals maintained high viral loads, and five of six developed fatal immunodeficiency during ∼80 weeks of follow-up. Lack of Vpr was associated with better preservation of CD4+ T cells, lower viral loads, and an attenuated clinical course of infection in most animals. Our results show that Vpr contributes to efficient viral immune evasion and the full pathogenic potential of SIVmac in vivo. Inhibition of Vpr may improve humoral immune control of viral replication.
    Keywords Immunology ; Immunity ; Immune response ; Science ; Q
    Subject code 570
    Language English
    Publishing date 2023-12-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

    Full text online

    More links

    Kategorien

    Inter-library loan at ZB MED

    Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

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