Immunology

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    Protocols in Current Issue
    Isolation and High Throughput Flow Cytometric Apoptosis Assay of Human Neutrophils to Enable Compound Library Screening
    Authors:  K. D. Herman, A. Rahman and L. R. Prince, date: 06/05/2020, view: 26, Q&A: 0
    The study of human neutrophils in vitro is challenging due to their short half-life and propensity for activation. However, with careful handling and manipulation in the laboratory, they can be a powerful tool to investigate immune ...
    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: 29, Q&A: 0
    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 ...
    RNA Extraction from Ears and Draining Lymph Nodes of Mice Infected with Leishmania amazonensis
    Authors:  Emilie Giraud and Evie Melanitou, date: 06/05/2020, view: 25, Q&A: 0
    Parasites of the genus Leishmania infect the mammalian hosts, including mice and humans and cause cutaneous or visceral leishmaniasis depending upon the parasite species transmitted by the vector sandfly. Leishmania amazonensis is ...
    Isolation and High Throughput Flow Cytometric Apoptosis Assay of Human Neutrophils to Enable Compound Library Screening
    Authors:  K. D. Herman, A. Rahman and L. R. Prince, date: 06/05/2020, view: 26, Q&A: 0
    [Abstract] The study of human neutrophils in vitro is challenging due to their short half-life and propensity for activation. However, with careful handling and manipulation in the laboratory, they can be a powerful tool to investigate immune responses in health and disease. Here we describe a method for the isolation of human neutrophils from ...
    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: 29, 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 ...
    RNA Extraction from Ears and Draining Lymph Nodes of Mice Infected with Leishmania amazonensis
    Authors:  Emilie Giraud and Evie Melanitou, date: 06/05/2020, view: 25, Q&A: 0
    [Abstract] Parasites of the genus Leishmania infect the mammalian hosts, including mice and humans and cause cutaneous or visceral leishmaniasis depending upon the parasite species transmitted by the vector sandfly. Leishmania amazonensis is one of the Leishmania species responsible for the cutaneous form of the disease. We have ...
    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: 638, 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 ...
    Direct Reprogramming of Mouse Embryonic Fibroblasts to Conventional Type 1 Dendritic Cells by Enforced Expression of Transcription Factors
    Authors:  Fábio F. Rosa, Cristiana F. Pires, Olga Zimmermannova and Carlos-Filipe Pereira, date: 05/20/2020, view: 611, Q&A: 0
    [Abstract] Ectopic expression of transcription factor combinations has been recently demonstrated to reprogram differentiated somatic cells towards the dendritic cell (DC) lineage without reversion to a multipotent state. DCs have the ability to induce potent and long-lasting adaptive immune responses. In particular, conventional type 1 DCs (cDC1s) excel on ...
    Bacterial Lawn Avoidance and Bacterial Two Choice Preference Assays in Caenorhabditis elegans
    Authors:  Jogender Singh and Alejandro Aballay, date: 05/20/2020, view: 427, Q&A: 0
    [Abstract] Physical avoidance of pathogens is a crucial defense strategy used by the host to reduce pathogen infection. Hosts display the use of multiple strategies to sense and avoid pathogens, ranging from olfaction to sensing of damage caused by pathogen infection. Understanding various mechanisms of pathogen avoidance has the potential to uncover ...
    Evaluation of B Cell Proliferation in vivo by EdU Incorporation Assay
    Authors:  Adi Biram and Ziv Shulman, date: 05/05/2020, view: 592, 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 ...
    HIV-CRISPR: A CRISPR/Cas9 Screening Method to Identify Genes Affecting HIV Replication
    Authors:  Ferdinand Roesch and Molly OhAinle, date: 05/05/2020, view: 549, Q&A: 0
    [Abstract] Screening with CRISPR/Cas9 technology has already led to significant discoveries in the fields of cancer biology, cell biology and virology. Because of the relatively low false discovery rates and the ability to perform high-throughput, pooled approaches, it has rapidly become the assay of choice for screening studies, including whole-genome ...
    Assessing in vitro and in vivo Trogocytosis By Murine CD4+ T cells
    Authors:  Jim Reed and Scott A. Wetzel, date: 05/05/2020, view: 554, 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 ...
    Environmental Conditioning and Aerosol Infection of Mice
    Authors:  Eriko Kudo and Akiko Iwasaki, date: 04/20/2020, view: 554, Q&A: 0
    [Abstract] Influenza infection models in mice are widely used to study flu-mediated immune responses and pathology. However, most laboratory mice are housed at 20 °C and 50% relative humidity (RH). To better recapitulate influenza epidemics and immune responses during winter seasons, mice were housed at 20 °C under different humidity conditions, 10-20% or ...



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