Immunology

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    Protocols in Current Issue
    Multiplex T-cell Stimulation Assay Utilizing a T-cell Activation Reporter-based Detection System
    Authors:  Laurie G. Landry, Sarah E. Mann, Amanda M. Anderson and Maki Nakayama, date: 01/20/2021, view: 87, Q&A: 0
    [Abstract]

    Immune tolerance and response are both largely driven by the interactions between the major histocompatibility complex (MHC) expressed by antigen presenting cells (APCs), T-cell receptors (TCRs) on T-cells, and their cognate antigens. Disordered interactions cause the pathogenesis of autoimmune diseases such as type 1 diabetes. Therefore, the

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    Analysis of B Cell Migration by Intravital Microscopy
    [Abstract]

    During immune responses, B cells home to lymph nodes (LN), where they encounter antigens. Homing starts with capture and L-selectin-dependent rolling on the activated endothelium of high endothelial venules (HEV). After recognition of chemokines presented on HEV, activation of B cell integrins occurs mediating firm arrest. Subsequently, B cells

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    Protocol for Isolation, Stimulation and Functional Profiling of Primary and iPSC-derived Human NK Cells
    [Abstract]

    Natural killer (NK) cells are innate immune cells, characterized by their cytotoxic capacity, and chemokine and cytokine secretion upon activation. Human NK cells are identified by CD56 expression. Circulating NK cells can be further subdivided into the CD56bright (~10%) and CD56dim NK cell subsets (~90%). NK cell-like cells can also be derived

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    Magnet-assisted Flow Cytometry of in vivo Tumors to Quantitate Cell-specific Responses to Magnetic Iron Oxide Nanoparticles
    Authors:  Preethi Korangath and Robert Ivkov, date: 11/20/2020, view: 678, Q&A: 0
    [Abstract] A clear understanding of nanoparticle interactions with living systems at the cellular level is necessary for developing nanoparticle-based therapeutics. Magnetic iron oxide nanoparticles provide unique opportunities to study these interactions because of their responsiveness to magnetic fields. This enables sorting of cells containing ...
    Flow-cytometric Detection of Low-level Reactive Oxygen Species in Cell Lines and Primary Immune Cells
    Authors:  Kevin Bode, Corinna Link, Peter H. Krammer and Heiko Weyd, date: 09/05/2020, view: 2128, Q&A: 0
    [Abstract] Depending on its concentration and cellular origin the production of reactive oxygen species (ROS) in the organism serves a variety of functions. While high concentrations during an oxidative burst are used to fight pathogens, low to moderate amounts of ROS act as signaling molecules important for several physiological processes such as regulation ...
    Optogenetic Tuning of Protein-protein Binding in Bilayers Using LOVTRAP
    Authors:  Doug Tischer and Orion D. Weiner, date: 09/05/2020, view: 1890, Q&A: 0
    [Abstract] Modern microscopy methods are powerful tools for studying live cell signaling and biochemical reactions, enabling us to observe when and where these reactions take place from the level of a cell down to single molecules. With microscopy, each cell or molecule can be observed both before and after a given perturbation, facilitating better inference ...
    Neutrophil Extracellular Trap Killing Assay of Candida albicans
    Authors:  Sheng-Yang Wu and Betty A. Wu-Hsieh, date: 08/20/2020, view: 1236, Q&A: 0
    [Abstract] Fungal pathogen Candida albicans is one of the top leading causes of overall healthcare-associated bloodstream infections worldwide. Neutrophil is the major effector cell to clear C. albicans infection. Our study showed that mouse neutrophils utilize two independent mechanisms to kill C. albicans: one is CR3 downstream ...
    Co-immunostaining of ICAM-1, ICAM-2, and CD31 in Mouse Kidney Glomeruli
    Author:  Sun-sang J. Sung, date: 07/05/2020, view: 1072, Q&A: 0
    [Abstract] Glomerulonephritis (GN) is a common pathological condition in chronic kidney diseases that often leads to end stage renal failure. Mac-1 (CD11b/CD18)-mediated neutrophil, macrophage, and dendritic cell glomerular infiltration leading to cellular dysfunction and destruction is an important disease mechanism. The cellular distribution and dynamics ...
    Methodology for in vitro Assessment of Human T Cell Activation and Blockade
    Authors:  Joseph R. Podojil, Igal Ifergan, Ming-Yi Chiang, Joshua J. Meeks and Stephen D. Miller, date: 06/05/2020, view: 1429, Q&A: 0
    [Abstract] Methods to test both the functionality and mechanism of action for human recombinant proteins and antibodies in vitro have been limited by multiple factors. To test the functionality of a recombinant protein or antibody, the receptor, the receptor-associated ligand, or both must be expressed by the cells present within the in vitro ...
    Evaluation of B Cell Proliferation in vivo by EdU Incorporation Assay
    Authors:  Adi Biram and Ziv Shulman, date: 05/05/2020, view: 1617, Q&A: 0
    [Abstract] Generation of antibodies is crucial for establishing enduring protection from invading pathogens, as well as for maintaining homeostasis with commensal bacteria at mucosal surfaces. Chronic exposure to microbiota- and dietary- derived antigens results in continuous production of antibody producing cells within the Peyer’s patch germinal center ...



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