Immunology

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    Protocols in Current Issue
    Metabolomic and Lipidomic Analysis of Bone Marrow Derived Macrophages
    Authors:  Gretchen L. Seim, Steven V. John and Jing Fan, date: 07/20/2020, view: 1034, Q&A: 0
    [Abstract] Macrophages are highly plastic immune cells that are capable of adopting a wide array of functional phenotypes in response to environmental stimuli. The changes in macrophage function are often supported and regulated by changes in cellular metabolism. Capturing a comprehensive picture of metabolism is vital for understanding the role of metabolic ...
    Generation of T cells from Human and Nonhuman Primate Pluripotent Stem Cells
    [Abstract] Pluripotent stem cells (PSCs) have the potential to provide homogeneous cell populations of T cells that can be grown at a clinical scale and genetically engineered to meet specific clinical needs. OP9-DLL4, a stromal line ectopically expressing the Notch ligand Delta-like 4 (DLL4) is used to support differentiation of PSCs to T-lymphocytes. This ...
    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: 1135, 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 ...
    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: 1718, 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: 1524, 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 ...
    Isolation and Stimulation of Peritoneal Macrophages with Apoptotic Jurkat Cells to Produce IL-10
    Authors:  Mei Song and Xiaojing Ma, date: 12/20/2019, view: 1924, Q&A: 0
    [Abstract] Clearance of apoptotic cells by macrophages is critical to ensuring cellular homeostasis and suppression of autoimmunity. Macrophage recognition of apoptotic cells triggers an anti-inflammatory response, which is mediated by the release of IL-10, TGF-β etc. with concurrent inhibition of pro-inflammatory cytokines (such as TNFα, IL-12, ...
    Isolation of Neutrophil Nuclei for Use in NETosis Assays
    Author:  Dave Boucher, date: 09/05/2019, view: 2233, Q&A: 0
    [Abstract] Neutrophils are critical immune cells that protect our body against invading pathogens. They generate antibacterial DNA structures called neutrophil extracellular traps (NET). Recently we identified a new mechanism that enables NET formation. We observed that following recognition of lipopolysaccharides, inflammatory caspases cleave Gasdermin D ...
    Isolation and Long-term Cultivation of Mouse Alveolar Macrophages
    [Abstract] Alveolar macrophages (AM) are tissue-resident macrophages that colonize the lung around birth and can self-maintain long-term in an adult organism without contribution of monocytes. AM are located in the pulmonary alveoli and can be harvested by washing the lungs using the method of bronchoalveolar lavage (BAL). Here, we compared different ...
    TZA, a Sensitive Reporter Cell-based Assay to Accurately and Rapidly Quantify Inducible, Replication-competent Latent HIV-1 from Resting CD4+ T Cells
    Authors:  Anwesha Sanyal, Vatsala S. Rangachar and Phalguni Gupta, date: 05/20/2019, view: 3745, Q&A: 1
    [Abstract] The latent HIV-1 viral reservoir in resting CD4+ (rCD4+) T cells represents a major barrier to an HIV-1 cure. There is an ongoing effort to identify therapeutic approaches that will eliminate or reduce the size of this reservoir. However, clinical investigators lack an assay to determine whether or not a decrease in the ...
    Isolation and Culture of Mouse Lung ILC2s
    Authors:  Pauline Schmitt, Anais Duval, Emilie Mirey, Jean-Philippe Girard and Corinne Cayrol, date: 10/05/2018, view: 4765, Q&A: 0
    [Abstract] Group 2 Innate Lymphoid Cells (ILC2) play an important role in immune responses at barrier surfaces, notably in the lung during airway allergic inflammation or asthma. Several studies have described methods to isolate ILC2s from wild-type naive mice, most of them using cell sorting to obtain a pure population. Here, we describe in detail, a ...



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