Mammalia

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    Protocols in Current Issue
    CD8 T Cell Virus Inhibition Assay Protocol
    [Abstract]

    The human immunodeficiency virus (HIV)-1 viral inhibition assay (VIA) measures CD8+ T cell-mediated inhibition of HIV replication in CD4+ T cells and is increasingly used for clinical testing of HIV vaccines and immunotherapies. Different VIAs that differ in length of CD8:CD4 T cell culture periods (6–13 days), purity of CD4 cultures

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    Restimulation-Induced Cell Death (RICD): Methods for Modeling, Investigating, and Quantifying RICD Sensitivity in Primary Human T Cells via Flow Cytometric Analysis
    Authors:  Katherine Pohida, Camille M. Lake, Debra Yee and Andrew L. Snow, date: 02/20/2022, view: 1077, Q&A: 0
    [Abstract]

    When the body mounts an immune response against a foreign pathogen, the adaptive arm of the immune system relies upon clonal expansion of antigen-specific T cell populations to exercise acquired effector and cytotoxic functions to clear it. However, T cell expansion must be modulated to effectively combat the perceived threat without inducing

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    HiSAT: A Novel Method for the Rapid Diagnosis of Allergy
    Authors:  Hirotomo Shibaguchi and Yuki Yasutaka, date: 02/05/2022, view: 895, Q&A: 0
    [Abstract]

    To identify causative substances for allergies to drugs or foods, the lymphocyte transformation test (LTT) is currently widely used as in vitro test, but its accuracy is not satisfactory. We have developed a novel method designated high-sensitivity allergy test (HiSAT) for determining allergy expression by measuring cell kinetics, using the

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    Static Adhesion Assay for Human Peripheral Blood Mononuclear Cells
    Authors:  Giulia Vanoni, Camilla Jandus and Sara Trabanelli, date: 01/05/2022, view: 1305, Q&A: 0
    [Abstract]

    Blood endothelial cells (ECs) constitute the primary physical barrier to be crossed by circulating leukocytes, once attracted to a site of ongoing inflammation/infection. Upon a pro-inflammatory stimulus, such as tumor necrosis factor (TNF), ECs upregulate adhesion molecule expression to favor the adhesion and, subsequently, the transendothelial

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    Evaluating Human Natural Killer Cells Antibody-dependent Cellular Cytotoxicity (ADCC) Using Plate-bound Anti-CD16 Antibodies
    Author:  Weiru Liu, date: 01/05/2022, view: 1236, Q&A: 0
    [Abstract]

    Natural killer (NK) cells are large granular lymphocytes that keep in check the health of neighboring cells through a large array of intrinsically expressed germline-coded receptors. Most importantly, CD16 is a low affinity Fc receptor for IgG that mediates the antibody-dependent cellular cytotoxicity (ADCC) of NK cells, bridging the innate and

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    Measuring Total Classical Pathway and Activities of Individual Components of the Mouse Complement Pathway
    Author:  Wioleta M. Zelek, date: 10/05/2021, view: 1053, Q&A: 0
    [Abstract]

    The complement system is a central component of innate immunity, responsible for recognition and killing of bacteria by tagging invaders through opsonisation, thereby promoting phagocytosis, and by direct lysis. Complement activity is routinely measured using functional assays that utilise erythrocytes as targets. The classical pathway haemolytic

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    In vitro and In vivo CD8+ T Cell Suppression Assays
    Authors:  Lu Xie, Guihuan Liu, Yanjun Liu and Yuming Yu, date: 05/20/2021, view: 3292, Q&A: 0
    [Abstract]

    CD8+CD28− T suppressor cells (Ts) have been documented to promote immune tolerance by suppressing effector T cell responses to alloantigens following transplantation. The suppressive function of T cells has been defined as the inhibitory effect of Ts on the proliferation rate of effector T cells. 3H-thymidine is a classical immunological

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    An Imaging Flow Cytometry Method to Measure Citrullination of H4 Histone as a Read-out for Neutrophil Extracellular Traps Formation
    Authors:  Emilia A. Barbu, Venina M. Dominical, Laurel Mendelsohn and Swee Lay Thein, date: 02/20/2021, view: 2559, Q&A: 0
    [Abstract]

    The formation of neutrophil extracellular traps (NETs) is thought to play a critical role in infections and propagating sterile inflammation. Histone citrullination is an essential and early step in NETs formation, detectable prior to the formation of the hallmark extracellular DNA-scaffolded strands. In addition to the classical microscopy

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    A Versatile Protocol to Quantify BCR-mediated Phosphorylation in Human and Murine B Cell Subpopulations
    Authors:  Jasper Rip, Rudi W. Hendriks and Odilia B. J. Corneth, date: 02/05/2021, view: 1725, Q&A: 0
    [Abstract]

    Signal transduction is the process by which molecular signals are transmitted from the cell surface to its interior, resulting in functional changes inside the cell. B cell receptor (BCR) signaling is of crucial importance for B cells, as it regulates their differentiation, selection, survival, cellular activation and proliferation. Upon BCR

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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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