Immunology

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    Protocols in Current Issue
    Murine Double Hit Model for Neonatal Cardiopulmonary Diseases: Bronchopulmonary Dysplasia (BPD) and Pulmonary Hypertension Associated with BPD
    Authors:  Steven P. Garrick, Philip J. Berger, Marcel F. Nold and Claudia A. Nold-Petry, date: 11/05/2022, view: 443, Q&A: 0
    [Abstract]

    Bronchopulmonary dysplasia (BPD) and pulmonary hypertension associated with BPD (BPD-PH) are of multifactorial origin and share common risk factors. Most murine models of BPD expose newborn pups to only one of these risk factors—more commonly postnatal hyperoxia—thereby mimicking the vital increased fraction of inspired oxygen (FiO2)

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    Flow Cytometry Analysis of SIRT6 Expression in Peritoneal Macrophages
    [Abstract]

    The sirtuin 6 has emerged as a regulator of acute and chronic immune responses. Recent findings show that SIRT6 is necessary for mounting an active inflammatory response in macrophages. In vitro studies revealed that SIRT6 is stabilized in the cytoplasm to promote tumor necrosis factor (TNFα) secretion. Notably, SIRT6 also promotes TNFα secretion

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    High-Efficiency Retroviral Transduction for Type 1 Regulatory T Cell Differentiation
    Authors:  Michael C. McGee, Avery August and Weishan Huang, date: 09/05/2022, view: 708, Q&A: 0
    [Abstract]

    Type 1 regulatory T (Tr1) cells are an immunoregulatory CD4+ Foxp3- IL-10high T cell subset with therapeutic potential for various inflammatory diseases. Retroviral (RV) transduction has been a valuable tool in defining the signaling pathways and transcription factors that regulate Tr1 differentiation and suppressive function. This protocol

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    Combination of Sterile Injury and Microbial Contamination to Model Post-surgical Peritoneal Adhesions in Mice
    Authors:  Julia Bayer, Deborah Stroka, Paul Kubes, Daniel Candinas and Joel Zindel, date: 08/20/2022, view: 1235, Q&A: 0
    [Abstract]

    Abdominal surgeries are frequently associated with the development of post-surgical adhesions. These are irreversible fibrotic scar bands that appear between abdominal organs and the abdominal wall. Patients suffering from adhesions are at risk of severe complications, such as small bowel obstruction, chronic pelvic pain, or infertility. To date,

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    CRISPR/Cas9-mediated Gene Knockout Followed by Negative Selection Leads to a Complete TCR Depletion in orthoCAR19 T Cells
    [Abstract]

    Genome-editing technologies, especially CRISPR (clustered regularly interspaced short palindrome repeats)/Cas9 (CRISPR-associated protein 9), endows researchers the ability to make efficient, simple, and precise genomic DNA changes in many eukaryotic cell types. CRISPR/Cas9-mediated efficient gene knockout holds huge potential to improve the

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    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: 673, 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: 1412, 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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    Serological Measurement of Poly-IgA Immune Complex Levels in IgA Nephropathy and IgA Vasculitis
    Authors:  Xue Zhang, Jicheng Lv, Pan Liu, Xinfang Xie, Xinyan Li, Hong Zhang and Jing Jin, date: 07/05/2022, view: 1555, Q&A: 0
    [Abstract]

    Both IgA nephropathy and IgA vasculitis, formerly known as Henoch-Schӧnlein purpura, are immune deposition diseases. IgA nephropathy is caused by the deposition of aberrantly formed poly-IgA complexes from blood circulation to the kidney glomerulus; IgA vasculitis is characterized by IgA-dominant immune deposits to small vessels of the skin

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    Protocol for High Throughput Screening of Antibody Phage Libraries
    [Abstract]

    Phage display is a proven and widely used technology for selecting specific antibodies against desired targets. However, an immense amount of effort is required to identify and screen the desired positive clones from large and diverse combinatorial libraries. On the other hand, the selection of positive binding clones

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    Protocol to Isolate Germinal Centers by Laser Microdissection
    Authors:  Farbod Bahreini, Markus Niebuhr, Julia Belde, Jürgen Westermann and Kathrin Kalies, date: 06/05/2022, view: 991, Q&A: 0
    [Abstract]

    During adaptive immune responses, germinal centers (GC) appear as transient microstructures, in which antigen-specific B and T cells interact with each other. Because only the antigen-activated B and T cells, such as Plasmablasts or follicular T helper (Tfh) cells, are present in GC, the in depth-analysis of GC is of great interest. To identify

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