Immunology

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    Protocols in Current Issue
    Cost Effective Method for gDNA Isolation from the Cecal Content and High Yield Procedure for RNA Isolation from the Colonic Tissue of Mice
    Authors:  Sohini Mukhopadhyay and Palok Aich, date: 08/05/2022, view: 193, Q&A: 0
    [Abstract]

    Microbiome studies are quickly gaining momentum. Since most of the resident microbes (consisting of bacteria, fungi, and viruses) are difficult to culture, sequencing the microbial genome is the method of choice to characterize them. It is therefore important to have efficient methodology for gDNA isolation of gut microbes. Mouse models are widely

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    Immunohistochemistry of Immune Cells and Cells Bound to in vivo Administered Antibodies in Liver, Lung, Pancreas, and Colon of B6/lpr Mice
    Authors:  Kieran Adam and Adam Mor, date: 07/20/2022, view: 787, Q&A: 0
    [Abstract]

    Employing a novel mouse model of immune related adverse events (irAEs) induced by combination of anti-PD1 and anti-CTLA-4 antibodies, we visualized immune infiltration into the liver, lung, pancreas, and colon. Here, we describe the avidin-biotin conjugate (ABC) method used to stain T cells (CD4 and CD8), B cells (CD19), macrophages (F4/80), and

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    Assessing the Presence of Hematopoietic Stem and Progenitor Cells in Mouse Spleen
    Authors:  Isabelle J. Marié, Lara Brambilla and David E. Levy, date: 06/05/2022, view: 899, Q&A: 0
    [Abstract]

    Transplantation of hematopoietic material into recipient mice is an assay routinely used to determine the presence and function of hematopoietic stem and progenitor cells (HSPCs) in vivo. The principle of the method is to transplant donor cells being tested for HSPCs into a recipient mouse following bone marrow ablation and testing for

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    Protocol to Induce Follicular T Helper Cells, Germinal Centers, and Skin Lesions in Mouse Models for Skin Blistering Diseases
    [Abstract]

    Autoreactive T cells in autoantibody-mediated autoimmune diseases can be divided into two major subsets: (i) follicular T helper cells (Tfh) that provide T cell help in germinal centers (GC) and (ii) effector T (Teff) cells that immigrate into peripheral tissue sites such as the skin and mediate local inflammation. To study the sequence of events

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    A Combinatorial in-silico, in-vitro and in-vivo Approach to Quantitatively Study Peptide Induced MHC Stability
    Authors:  Roman Sarkar, Yashu Sharma, Ayush Jain, Azeez Tehseen, Sudhakar Singh and Sharvan Sehrawat, date: 12/20/2021, view: 1238, Q&A: 0
    [Abstract]

    Here, we describe a combinatorial approach in reverse vaccinology to identify immunogenic class I major histocompatibility complex (MHC) displayed epitopes derived from a morbillivirus named pestes des petits ruminants (PPRV). The protocol describes an in silico prediction of immunogenic epitopes using an IEDB tool. The predicted peptides were

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    Intact in situ Preparation of the Drosophila melanogaster Lymph Gland for a Comprehensive Analysis of Larval Hematopoiesis
    Authors:  Diana Rodrigues, K. VijayRaghavan, Lucas Waltzer and Maneesha S. Inamdar, date: 11/05/2021, view: 2243, Q&A: 0
    [Abstract]

    Blood cells have a limited lifespan and are replenished by a small number of hematopoietic stem and progenitor cells (HSPCs). Adult vertebrate hematopoiesis occurs in the bone marrow, liver, and spleen, rendering a comprehensive analysis of the entire HSPC pool nearly impossible. The Drosophila blood system is well studied and has developmental,

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    Isolation and Culturing Primary Cholangiocytes from Mouse Liver
    [Abstract]

    Cholangiocytes are epithelial cells lining the intrahepatic and extrahepatic bile ducts. Cholangiocytes perform key physiological functions in the liver. Bile synthesized by hepatocytes is secreted into bile canaliculi, further stored in the gallbladder, and finally discharged into the duodenum. Due to liver injury, biliary epithelial proliferate

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    Isolation of Nasal Brush Cells for Single-cell Preparations
    Authors:  Saltanat Ualiyeva, Amelia A. Boyd, Nora A. Barrett and Lora G. Bankova, date: 09/20/2021, view: 2416, Q&A: 0
    [Abstract]

    Solitary chemosensory epithelial cells are scattered in most mucosal surfaces. They are referred to as tuft cells in the intestinal mucosa, brush cells in the trachea, and solitary chemosensory and microvillous cells in the nasal mucosa. They are the primary source of IL-25 in the epithelium and are also engaged in acetylcholine generation. We

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    Parasitemia Evaluation in Mice Infected with Schistosoma mansoni
    Authors:  Naiara Clemente Tavares and Marina Moraes Mourão, date: 05/20/2021, view: 2868, Q&A: 0
    [Abstract]

    Schistosomiasis is a neglected tropical disease. Its treatment relies on the use of a single drug, praziquantel. Due to treatment limitations, an alternative for schistosomiasis chemotherapy is required; thus, a better understanding of parasite biology and host-parasite interactions is valuable to aid the identification of new anti-Schistosoma

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    In vivo CD40 Silencing by siRNA Infusion in Rodents and Evaluation by Kidney Immunostaining
    [Abstract]

    The co-stimulatory molecule CD40 and its ligand CD40L play a key role in the regulation of immunological processes and are involved in the pathophysiology of autoimmune and inflammatory diseases. Inhibition of the CD40-CD40L axis is a promising therapy, and a number of strategies and techniques have been designed to hinder its functionality. Our

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