Immunology

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    Protocols in Current Issue
    Characterization of the Elasticity of CD4+ T Cells: An Approach Based on Peak Force Quantitative Nanomechanical Mapping
    Authors:  Philipp Jung, Xiangda Zhou, Sandra Iden, Bin Qu and Markus Bischoff, date: 04/20/2022, view: 752, Q&A: 0
    [Abstract]

    CD4+ T cells are essential players in orchestrating the specific immune response against intracellular pathogens, and in inhibiting tumor development in an early stage. The activation of T cells is triggered by engagement of T cell receptors (TCRs). Here, CD3 and CD28 molecules are key factors, (co)stimulating signaling pathways essential for

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    Differentiation of Human Induced Pluripotent Stem Cell into Macrophages
    Authors:  Harriet Douthwaite, Aitor Bermejo Arteagabeitia and Subhankar Mukhopadhyay, date: 03/20/2022, view: 862, Q&A: 0
    [Abstract]

    As a model to interrogate human macrophage biology, macrophages differentiated from human induced pluripotent stem cells (hiPSCs) transcend other existing models by circumventing the variability seen in human monocyte-derived macrophages, whilst epitomizing macrophage phenotypic and functional characteristics over those offered by macrophage-like

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    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: 1075, 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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    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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    Bacterial Infection with Listeria monocytogenes in Mice and Subsequent Analysis of Antigen-Specific CD8 T Cell Responses
    Authors:  Bojana Jakic, Janine Kimpel, William J. Olson, Verena Labi and Natascha Hermann-Kleiter, date: 12/05/2021, view: 1187, Q&A: 0
    [Abstract]

    Pathogens such as bacteria, viruses, fungi, or protozoa can cause acute and chronic infections in their hosts. The intracellular bacterium Listeria monocytogenes serves as a model pathogen to assess the molecular mechanisms regulating CD8 T cell activation, differentiation, and function. We set up an experimental workflow to investigate

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    Lentivirus-mediated Conditional Gene Expression
    [Abstract]

    The ability to identify the role of a particular gene within a system is dependent on control of the expression of that gene. In this protocol, we describe a method for stable, conditional expression of Nod-Like receptors (NLRs) in THP-1 cells using a lentiviral expression system. This system combines all the necessary components for

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    Microscopic Detection of ASC Inflammasomes in Bone Marrow Derived Macrophages Post Stimulation
    Authors:  Longjun Li, Rudi Mao and Xiaopeng Qi, date: 09/05/2021, view: 1684, Q&A: 0
    [Abstract]

    An inflammasome is an intracellular multiprotein complex that plays important roles in host defense and inflammatory responses. Inflammasomes are typically composed of the adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC), cytoplasmic sensor protein, and the effector protein pro-caspase-1. ASC assembly into a protein

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    A Lipidomics Approach to Measure Phosphatidic Acid Species in Subcellular Membrane Fractions Obtained from Cultured Cells
    Authors:  Nawal Kassas, Laetitia Fouillen, Stéphane Gasman and Nicolas Vitale, date: 06/20/2021, view: 1862, Q&A: 0
    [Abstract]

    Over the last decade, lipids have emerged as possessing an ever-increasing number of key functions, especially in membrane trafficking. For instance, phosphatidic acid (PA) has been proposed to play a critical role in different steps along the secretory pathway or during phagocytosis. To further investigate in detail the precise nature of PA

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