Immunology

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    Protocols in Current Issue
    Intestinal Enteroid Culture for Human Astroviruses
    Authors:  Irene A. Owusu, Carmen Mirabelli and Abimbola O. Kolawole, date: 07/20/2020, view: 239, Q&A: 0
    [Abstract] Human astroviruses (HAstV) are non-enveloped, positive-sense single stranded RNA viruses that typically cause gastroenteritis in children, the elderly and among immunocompromised individuals. Some HAstV species have also been implicated in neurological diseases. It is important to study these viruses to understand the pathogenesis and develop ...
    Zebrafish Bacterial Infection Assay to Study Host-Pathogen Interactions
    Authors:  Faiza Basheer, Clifford Liongue and Alister C Ward, date: 03/05/2020, view: 995, Q&A: 0
    [Abstract] The study of host-pathogen interactions has improved our understanding of both pathogenesis and the response of the host to infection, including both innate and adaptive responses. Neutrophils and macrophages represent the first line of innate host defense against any infection. The zebrafish is an ideal model to study the response of these cells ...
    Preparation of Red Palm Weevil Rhynchophorus Ferrugineus (Olivier) (Coleoptera: Dryophthoridae) Germ-free Larvae for Host-gut Microbes Interaction Studies
    Authors:  Abrar Muhammad, Prosper Habineza, Youming Hou and Zhanghong Shi, date: 12/20/2019, view: 1494, Q&A: 0
    [Abstract] Red palm weevil (RPW), Rhynchophorus ferrugineus Olivier, is a devastating pest of palm trees worldwide. RPW gut is colonized by diverse bacterial species which profoundly influence host development and nutritional metabolism. However, the molecular mechanisms behind the interactions between RPW and its gut microbiota remain mostly ...
    Platelet Isolation and Activation Assays
    Authors:  Laura C. Burzynski, Nicholas Pugh and Murray C.H. Clarke, date: 10/20/2019, view: 2268, Q&A: 0
    [Abstract] Platelets regulate hemostasis and are the key determinants of pathogenic thrombosis following atherosclerotic plaque rupture. Platelets circulate in an inactive state, but become activated in response to damage to the endothelium, which exposes thrombogenic material such as collagen to the blood flow. Activation results in a number of responses, ...
    A Highly Sensitive Anion Exchange Chromatography Method for Measuring cGAS Activity in vitro
    Authors:  Andreas Holleufer and Rune Hartmann, date: 10/20/2018, view: 2557, Q&A: 0
    [Abstract] Cyclic GMP-AMP synthase (cGAS) is a pattern recognition receptor (PRR) that senses double stranded DNA (dsDNA) in the cytosol and this leads to the activation of stimulator of interferon genes (STING) via the secondary messenger 2’3’-cyclic GMP-AMP (2’3’-cGAMP). STING then recruits TANK binding kinase 1 (TBK-1) and this complex can phosphorylate ...
    Immunogold Electron Microscopy of the Autophagosome Marker LC3
    Authors:  Domenico Mattoscio, Andrea Raimondi, Carlo Tacchetti and Susanna Chiocca, date: 12/20/2017, view: 7588, Q&A: 0
    [Abstract] Even though autophagy was firstly observed by transmission electron microscopy already in the 1950s (reviewed in Eskelinen et al., 2011), nowadays this technique remains one of the most powerful systems to monitor autophagic processes. The autophagosome, an LC3-positive double membrane structures enclosing cellular materials, represents ...
    Detection of ASC Oligomerization by Western Blotting
    Authors:  Jérôme Lugrin and Fabio Martinon , date: 05/20/2017, view: 11486, Q&A: 0
    [Abstract] The apoptosis-associated speck-like protein with a caspase-recruitment domain (ASC) adaptor protein bridges inflammasome sensors and caspase-1. Upon inflammasome activation, ASC nucleates in a prion-like manner into a large and single platform responsible for the recruitment and the activation of caspase-1. Active caspase-1 will in turn promote ...
    Endogenous C-terminal Tagging by CRISPR/Cas9 in Trypanosoma cruzi
    Authors:  Noelia Lander, Miguel A. Chiurillo, Aníbal E. Vercesi and Roberto Docampo, date: 05/20/2017, view: 9529, Q&A: 0
    [Abstract] To achieve the C-terminal tagging of endogenous proteins in T. cruzi we use the Cas9/pTREX-n vector (Lander et al., 2015) to insert a specific tag sequence (3xHA or 3xc-Myc) at the 3’ end of a specific gene of interest (GOI). Chimeric sgRNA targeting the 3’ end of the GOI is PCR-amplified and cloned into Cas9/pTREX-n vector. Then ...
    Differential Salt Fractionation of Nuclei to Analyze Chromatin-associated Proteins from Cultured Mammalian Cells
    Authors:  Christin Herrmann, Daphne C. Avgousti and Matthew D. Weitzman, date: 03/20/2017, view: 10512, Q&A: 0
    [Abstract] Nucleosomes are the core units of cellular chromatin and are comprised of 147 base pairs (bp) of DNA wrapped around an octamer of histone proteins. Proteins such as chromatin remodelers, transcription factors, and DNA repair proteins interact dynamically with chromatin to regulate access to DNA, control gene transcription, and maintain genome ...
    Preparation of Protein-containing Extracts from Microbiota-rich Intestinal Contents
    [Abstract] The contribution of microbiota in regulating multiple physiological and pathological host responses has been studied intensively in recent years. Evidence suggests that commensal microbiota can directly modulate different populations of cells of the immune system (e.g., Ivanov et al., 2008; Atarashi et al., 2011). ...



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