Mammalia

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    Protocols in Current Issue
    Detection of Antigen-specific T cells in Spleens of Vaccinated Mice Applying 3[H]-Thymidine Incorporation Assay and Luminex Multiple Cytokine Analysis Technology
    Authors:  Maria Agallou and Evdokia Karagouni, date: 06/05/2019, view: 801, Q&A: 0
    [Abstract] For many infectious diseases T cells are an important part of naturally acquired protective immune responses, and inducing these by vaccination has been the aim of much research. Here, we describe a protocol for the analysis of vaccine-induced antigen-specific immune responses. For this purpose, cells of whole spleens obtained from vaccinated ...
    Induced Germinal Center B Cell Culture System
    Authors:  Kei Haniuda and Daisuke Kitamura, date: 02/20/2019, view: 2345, Q&A: 0
    [Abstract] The germinal center (GC) is the site where B cells undergo clonal expansion, affinity-based selection, and differentiation into memory B cells or plasma cells. It has been difficult to elucidate regulatory mechanisms for the dynamic GC B cell maturation and differentiation, partly because experimental manipulation of GC B cells in vivo ...
    ELISPOT Assay to Measure Peptide-specific IFN-γ Production
    Authors:  Michelle N. Wykes and Laurent Renia, date: 06/05/2017, view: 9043, Q&A: 0
    [Abstract] Interferon-gamma (IFN-γ) is crucial for immunity against intracellular pathogens and for tumor control. It is produced predominantly by natural killer (NK) and natural killer T cells (NKT) as well as by antigen-specific Th1 CD4+ and CD8+ effector T cells. When investigating immune responses against pathogens and cancer cells, ...
    Mouse CD8+ T Cell Migration in vitro and CXCR3 Internalization Assays
    Authors:  Rosa Barreira da Silva and Matthew L. Albert, date: 03/20/2017, view: 8393, Q&A: 0
    [Abstract] Chemokines are molecules that regulate the positioning of cells during homeostasis and inflammation. CXCL10 is an interferon-induced chemokine that attracts cells that express the chemokine receptor CXCR3 on their surface. CXCL10 expression is often induced upon inflammation and guides lymphocytes, such as T and NK cells, into the injured tissues. ...
    In vivo DCs Depletion with Diphtheria Toxin and MARCO+/MOMA1+ Cells Depletion with Clodronate Liposomes in B6.CD11c-DTR Mice
    [Abstract] To evaluate precisely the relative roles of different splenic phagocytic cells during an immune response, efficient methods for the depletion of specific populations are needed. Here, we describe the protocols for the depletion of splenic dendritic cells (DCs) by human diphtheria toxin (DTx) treatment in target mice (which express the human DTx ...
    T Cell Transfer Model of Colitis
    Authors:  Wen Pan, Shu Zhu, Dai Dai, Yuanjia Tang, Yihong Yao and Nan Shen, date: 07/05/2016, view: 9360, Q&A: 0
    [Abstract] Inflammatory bowel disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC) is mainly caused by disordered immune regulation and dysregulated intestinal microbiota. Here we present the T cell transfer model which has extensively used in many studies to identify the regulatory T cell function in gut inflammation. Naïve T cells only ...
    In vivo OVA-specific Cytotoxic CD8+ T Cell Killing Assay
    Authors:  Nada Chaoul, Catherine Fayolle and Claude Leclerc, date: 06/20/2016, view: 9637, Q&A: 0
    [Abstract] Cytotoxic CD8+ T cells are responsible for the lysis of cells expressing peptides associated with MHC class I molecules and derived from infection with a pathogen or from mutated antigens. In order to quantify in vivo this antigen-specific CD8+ T cell killing activity, we use the in vivo killing assay (IVK). ...
    Analysis of T Cell Proliferating and Polarizing Potential of Murine Dendritic Cells in Allogeneic-mixed Leukocyte Reaction
    Authors:  Pawan Kumar and Sangeeta Bhaskar, date: 03/05/2016, view: 6014, Q&A: 0
    [Abstract] Dendritic cells (DCs) play a critical role in mounting the T cell response against different infectious agents. Nature and intensity of the induced T cell responses are defined by activation status of DCs. It is generally accepted that IL-12, IL-4/IL-5 and IL-23 producing DCs induce TH1, TH2 and TH17 type of immune ...
    Protocol-In vitro T Cell Proliferation and Treg Suppression Assay with Celltrace Violet
    Authors:  Kristofor K. Ellestad and Colin C. Anderson, date: 01/05/2016, view: 18096, Q&A: 0
    [Abstract] Measurement of the incorporation of radionuclides such as 3H-thymidine is a classical immunological technique for assaying T cell proliferation. However, such an approach has drawbacks beyond the inconvenience of working with radioactive materials, such as the inability of bulk radionuclide incorporation measurements to accurately ...
    Generation of Aβ-specific T cell lines and in vivo Transfer
    Authors:  Róisín M. McManus, Marina A. Lynch and Kingston H.G. Mills, date: 09/20/2014, view: 6195, Q&A: 0
    [Abstract] Amyloid-β (Aβ)-containing plaques accumulate in the brains of patients with Alzheimer’s disease (AD). Studies in transgenic mice which over-express amyloid precursor protein and presenilin 1 (APP/PS1 mice) have suggested that T cells that infiltrate the brain may influence the development of Aβ plaques and associated cognitive dysfuncation. Active ...



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