Immunology

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    Protocols in Current Issue
    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: 452, Q&A: 0
    [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: 1658, 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 ...
    Adoptive Transfer of Tumor Expanded Regulatory T Cells (Tregs)
    Authors:  Felipe Vences-Catalán and Shoshana Levy, date: 08/20/2016, view: 5484, Q&A: 0
    [Abstract] Regulatory T cells (Tregs), a subset of CD4+CD25+ T cells, infiltrate tumors and suppress antitumor activity of effector T and NK cells. Depletion of Tregs by anti CD25+ antibodies has been shown to reduce tumor growth and metastasis (Olkhanud et al., 2009). Conversely, adoptive transfer of Tregs induced ...
    Isolation of Lymphocytes from Murine Visceral Adipose Tissue
    Authors:  Sonja Valentić, Felix M. Wensveen and Bojan Polić, date: 12/05/2015, view: 9311, Q&A: 1
    [Abstract] Several studies have shown that the detrimental influence of abdominal obesity on metabolic processes is mediated by the intra-abdominal fat depot. Visceral adipose tissue has been shown to be an independent risk factor for coronary heart disease, hypertension, impaired glucose tolerance and Diabetes Mellitus Type 2 (DM2). Diet-induced obesity in ...
    Adoptive Transfer of Memory B Cells
    Authors:  Griselda Zuccarino-Catania and Mark Shlomchik, date: 08/20/2015, view: 6281, Q&A: 0
    [Abstract] The adoptive transfer of antigen-specific B cells into mice that cannot recognize that specific antigen has two main advantages. The first is determining exactly when the B cells were transferred and exposed to antigen. The second is that all B cells that can bind that antigen are the ones that were transferred; no new antigen-specific B cells ...
    Separation of Intracellular Vesicles for Immunoassays
    Authors:  Toshihiko Kobayashi, Tsubasa Tanaka and Noriko Toyama-Sorimachi, date: 08/20/2015, view: 6201, Q&A: 0
    [Abstract] The endosome/lysosome systems play important roles in immune cell functions as signaling platforms. Immune cells utilize these endosome/lysosome for signal transduction or intercellular communication to elicit the proper immune responses, regulating the localization or the association of the signaling complexes. Here we introduce the procedures to ...
    Ex vivo Human Natural Killer (NK) Cell Stimulation and Intracellular IFNγ and CD107a Cytokine Staining
    Authors:  Vanessa A. York and Jeffrey M. Milush, date: 06/20/2015, view: 9330, Q&A: 1
    [Abstract] Natural killer (NK) cells comprise 5–20% of peripheral blood mononuclear cells (PBMC) in humans. In addition to their fundamental roles in the defense against viral infections and tumor surveillance, NK cells help shape adaptive immune responses through their production of cytokines. NK cells are traditionally identified as CD3neg, CD14 ...
    Extraction and Identification of T Cell Stimulatory Self-lipid Antigens
    [Abstract] Autoreactive T cells restricted to CD1 molecules and specific for endogenous lipids are abundant in human blood (de Jong et al., 2010; de Lalla et al., 2011). A few self-lipid molecules recognized by diverse individual T cell clones and accumulated within APCs following stress signals or cell transformation have been identified ...
    Isolation of Lung Infiltrating Cell in Mice
    Authors:  Eduardo P. Amaral, Elena B. Lassunskaia and Maria Regina D’Império-Lima, date: 02/20/2015, view: 11531, Q&A: 0
    [Abstract] Inflammatory lung diseases induce strong leukocyte recruitment into the organ, culminating in pneumonia area formation. Here, we describe the protocol for isolation of lung infiltrating cells. Using this assay, we analyzed the lung cell phenotyping by flow cytometry and spontaneous cytokine production by cultivating lung cells ex vivo ...
    Isolation of CNS-infiltrating and Resident Microglial Cells
    Authors:  Young-Hee Jin and Byung S. Kim, date: 01/20/2015, view: 10223, Q&A: 0
    [Abstract] Variety of immunological and biochemical studies associated with infection or inflammation in the central nervous system (CNS) utilize CNS-resident and/or infiltrating cells which were isolated from the CNS of naïve and affected mice in order to investigate the underlying mechanisms and the potential roles of the cell populations. Mechanical and ...



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