Immunology

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    Protocols in Current Issue
    Multi-color Flow Cytometry for Comprehensive Analysis of the Tumor Immune Infiltrate in a Murine Model of Breast Cancer
    Authors:  Ana S. Almeida, Miriam R. Fein and Mikala Egeblad, date: 06/05/2021, view: 598, Q&A: 0
    [Abstract]

    Flow cytometry is a popular laser-based technology that allows the phenotypic and functional characterization of individual cells in a high-throughput manner. Here, we describe a detailed procedure for preparing a single-cell suspension from mammary tumors of the mouse mammary tumor virus-polyoma middle T (MMTV-PyMT) and analyzing these cells by

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    A Simple and Robust Protocol for in vitro Differentiation of Mouse Non-pathogenic T Helper 17 Cells from CD4+ T Cells
    Authors:  Siwen Kang, Ruohan Wu and Ruoning Wang, date: 05/20/2021, view: 828, Q&A: 0
    [Abstract]

    Functional and mechanistic studies of CD4+ T cell lineages rely on robust methods of in vitro T cell polarization. Here, we report an optimized protocol for in vitro differentiation of a mouse non-pathogenic T helper 17 (TH17) cell lineage. Most of the previously established protocols require irradiated splenocytes as artificial antigen presenting

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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: 677, 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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    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: 2737, 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 ...
    Flow Cytometry Analysis and Fluorescence-activated Cell Sorting of Myeloid Cells from Lung and Bronchoalveolar Lavage Samples from Mycobacterium tuberculosis-infected Mice
    Authors:  Alissa C. Rothchild, Dat Mai, Alan Aderem and Alan H. Diercks, date: 05/20/2020, view: 2704, Q&A: 0
    [Abstract] Mycobacterium tuberculosis (Mtb) is transmitted by aerosol and can cause serious bacterial infection in the lung that can be fatal if left untreated. Mtb is now the leading cause of death worldwide by an infectious agent. Characterizing the early events of in vivo infection following aerosol challenge is critical for ...
    Assessing in vitro and in vivo Trogocytosis By Murine CD4+ T cells
    Authors:  Jim Reed and Scott A. Wetzel, date: 05/05/2020, view: 2018, Q&A: 0
    [Abstract] Recognition of antigens by lymphocytes (B, T, and NK) on the surface of an antigen-presenting cell (APC) leads to lymphocyte activation and the formation of an immunological synapse between the lymphocyte and the APC. At the immunological synapse APC membrane and associated membrane proteins can be transferred to the lymphocyte in a process called ...
    Characterization of Immunological Niches within Peyer’s Patches by ex vivo Photoactivation and Flow Cytometry Analysis
    Authors:  Adi Biram and Ziv Shulman, date: 03/20/2020, view: 2008, Q&A: 0
    [Abstract] T follicular helper (Tfh) cells regulate B cell selection for entry into the germinal center (GC) reaction or for differentiation into antibody forming cells. This process takes place at the border between the T and B zones in lymphoid organs and involves physical contacts between T and B cells. During these interactions, T cells endow the B cells ...
    Skin Transplantation and Lymphoid Organ Analysis in Mice
    [Abstract] Skin transplantation in mice is an important procedure to evaluate immune responses generated against heterologous grafts, especially given its highly immunogenic nature. In fact, skin is one of the most challenging organs in terms of allograft retention. In this protocol, we provide a detailed procedure for skin grafting using the tail skin as ...
    Immunohistochemical Staining of CD8α in Diabetic Mouse Kidney
    Authors:  Ming-Zhi Zhang and Raymond C. Harris, date: 09/20/2019, view: 1511, Q&A: 0
    [Abstract] Immune cell infiltration, particularly cytotoxic CD8α lymphocyte infiltration, plays an important role in development of diabetic nephropathy. Although CD8α infiltration can be evaluated by its production of cytokines, its localization in the kidney is of particular importance. The current protocol describes CD8α immunostaining using a Vectastain ...
    In vitro Infection of Primary Human Monocytes with HIV-1
    Authors:  Patrick O’Connell, Yong-Hui Zheng, Andrea Amalfitano and Yasser A. Aldhamen, date: 07/05/2019, view: 3107, Q&A: 0
    [Abstract] Monocyte infection by HIV-1 is an important component of chronic HIV pathogenesis. Following infection by HIV-1, monocytes are able to cross the blood brain barrier and set up a viral reservoir in the central nervous system. Additionally, in the setting of chronic HIV-1 infection, monocytes can become activated either directly through HIV-1 ...



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