Immunology

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    Protocols in Current Issue
    Gentamicin Protection Assay to Determine the Number of Intracellular Bacteria during Infection of Human TC7 Intestinal Epithelial Cells by Shigella flexneri
    Authors:  Atin Sharma and Andrea Puhar, date: 07/05/2019, view: 623, Q&A: 0
    [Abstract] Shigella flexneri is an intracellular bacterial pathogen that gains access to the gut epithelium using a specialized Type III Secretion System (T3SS). Various determinants mediating this invasive infection have been experimentally verified using the classical gentamicin protection assay presented here. In this assay epithelial cell lines ...
    Platelet Migration and Bacterial Trapping Assay under Flow
    Authors:  Shuxia Fan, Michael Lorenz, Steffen Massberg and Florian Gaertner, date: 09/20/2018, view: 1868, Q&A: 0
    [Abstract] Blood platelets are critical for hemostasis and thrombosis, but also play diverse roles during immune responses. We have recently reported that platelets migrate at sites of infection in vitro and in vivo. Importantly, platelets use their ability to migrate to collect and bundle fibrin (ogen)-bound bacteria accomplishing ...
    Establishment of a Human Cell Line Persistently Infected with Sendai Virus
    Authors:  Christopher Coakley, Cara Peter, Stephanie Fabry and Saurabh Chattopadhyay, date: 08/20/2017, view: 4192, Q&A: 0
    [Abstract] Interferon regulatory transcription factor 3 (IRF3) is a transcription factor that upon activation by virus infection promotes the synthesis of antiviral genes, such as the interferons (Hiscott, 2007). In addition to inducing genes, IRF3 triggers antiviral apoptosis by RIG-I-like receptor-induced IRF3 mediated pathway of apoptosis (RIPA), which is ...
    Screening for Novel Endogenous Inflammatory Stimuli Using the Secreted Embryonic Alkaline Phosphatase NF-κB Reporter Assay
    Authors:  Lorena Zuliani-Alvarez, Anna M. Piccinini and Kim S Midwood, date: 04/05/2017, view: 5066, Q&A: 0
    [Abstract] An immune response can be activated by pathogenic stimuli, as well as endogenous danger signals, triggering the activation of pattern recognition receptors and initiating signalling cascades that lead to inflammation. This method uses THP1-BlueTM cells, a human monocytic cell line which contains an embryonic alkaline phosphatase ...
    Isolation of Exosomes from Semen for in vitro Uptake and HIV-1 Infection Assays
    Authors:  Marisa N. Madison, Jennifer L. Welch and Chioma M. Okeoma, date: 04/05/2017, view: 5814, Q&A: 0
    [Abstract] Exosomes are membranous extracellular nanovesicles of endocytic origin. Exosomes are known to carry host and pathogen-derived genomic, proteomic, lipidomic cargos and other extraneous molecules. Exosomes are secreted by diverse cell types into the extracellular milieu and are subsequently internalized by recipient neighboring or distal cells. Upon ...
    Quantitative Measurements of HIV-1 and Dextran Capture by Human Monocyte-derived Dendritic Cells (MDDCs)
    Authors:  Mickaël M. Ménager and Dan R. Littman, date: 11/20/2016, view: 5413, Q&A: 0
    [Abstract] The aim of this protocol is to describe how to measure and quantify the amount of HIV-1 particles and dextran molecules internalized in human monocyte derived dendritic cells (MDDCs), using three different techniques: flow cytometry, quantitative PCR and confocal microscopy.
    Human Blood Component Vaccinia Virus Neutralization Assay
    Authors:  Laura Evgin and John Bell, date: 12/05/2015, view: 5160, Q&A: 0
    [Abstract] Many therapeutic viruses, such as oncolytic viruses, vaccines, or gene therapy vectors, may be administered by the intravenous route to maximize their delivery to target tissues. Blood components, such as antibody, complement and blood cells (such as neutrophils, monocytes, T cells, B cells or platelets) may result in viral neutralization and ...
    HIV-1 Fusion Assay
    Authors:  Marielle Cavrois, Jason Neidleman and Warner C. Greene, date: 08/20/2014, view: 10221, Q&A: 0
    [Abstract] The HIV-1 fusion assay measures all steps in the HIV-1 life cycle up to and including viral fusion. It relies on the incorporation of a β-lactamase–Vpr (BlaM-Vpr) protein chimera into the virion and the subsequent transfer of this chimera into the target cell by fusion (Figure 1). The transfer is monitored by the enzymatic cleavage of CCF2, a ...
    Luminescence-based Antiviral Assay for Hepatitis E Virus
    Authors:  Yannick Debing, Kai Dallmeier and Johan Neyts, date: 08/05/2014, view: 6445, Q&A: 0
    [Abstract] Hepatitis E virus (HEV) is one of the main causes of acute hepatitis worldwide. Infections are particularly severe in pregnant women and chronic hepatitis E is known to occur in immunocompromised patients. Current therapy (ribavirin or pegylated alpha interferon) has severe side effects and cannot be employed in all patients. In order to evaluate ...
    An ex vivo Model of HIV-1 Infection in Human Lymphoid Tissue and Cervico-vaginal Tissue
    Authors:  Andrea Introini, Christophe Vanpouille, Jean Charles Grivel and Leonid Margolis, date: 02/20/2014, view: 9658, Q&A: 0
    [Abstract] Human tissue explants are a valuable tool to study the interactions between host and infectious agents. They reliably mimic many important aspects of tissue cytoarchitecture and functions and allow us the investigation of the mechanisms of microbial pathogenesis under controlled laboratory conditions. One of the advantages of this system is that, ...



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