Immunology

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    Protocols in Current Issue
    In vitro and In vivo CD8+ T Cell Suppression Assays
    Authors:  Lu Xie, Guihuan Liu, Yanjun Liu and Yuming Yu, date: 05/20/2021, view: 2149, Q&A: 0
    [Abstract]

    CD8+CD28− T suppressor cells (Ts) have been documented to promote immune tolerance by suppressing effector T cell responses to alloantigens following transplantation. The suppressive function of T cells has been defined as the inhibitory effect of Ts on the proliferation rate of effector T cells. 3H-thymidine is a classical immunological

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    A Simple and Robust Protocol for in vitro Differentiation of Mouse Non-pathogenic T Helper 17 Cells from CD4+ T Cells
    Authors:  Siwen Kang, Ruohan Wu and Ruoning Wang, date: 05/20/2021, view: 1727, Q&A: 0
    [Abstract]

    Functional and mechanistic studies of CD4+ T cell lineages rely on robust methods of in vitro T cell polarization. Here, we report an optimized protocol for in vitro differentiation of a mouse non-pathogenic T helper 17 (TH17) cell lineage. Most of the previously established protocols require irradiated splenocytes as artificial antigen presenting

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    In vitro STING Activation with the cGAMP-STINGΔTM Signaling Complex
    Authors:  Yanpu He, Celestine Hong, Darrell J. Irvine, Jiahe Li and Paula T. Hammond, date: 02/05/2021, view: 2169, Q&A: 0
    [Abstract] Activating the STING (stimulator of interferon genes) signaling pathway via administration of STING agonist cyclic GMP-AMP (cGAMP) has shown great promise in cancer immunotherapy. While state-of-the-art approaches have predominantly focused on the encapsulation of cGAMP into liposomes or polymersomes for cellular delivery, we discovered that the ...
    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: 7147, 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 ...
    Imaging Cytokine Concentration Fields Using PlaneView Imaging Devices
    Authors:  Alon Oyler-Yaniv and Oleg Krichevsky, date: 04/05/2018, view: 4877, Q&A: 0
    [Abstract] We describe here a method to visualize concentration fields of cytokines around cytokine-secreting cells. The main challenge is that physiological cytokine concentrations can be very low, in the pico-molar range. Since it is currently impossible to measure such concentrations directly, we rely on cell’s response to the cytokines–the ...
    In vitro Homeostatic Proliferation of Human CD8 T Cells
    Authors:  Hossam A. Abdelsamed, Caitlin C. Zebley and Ben Youngblood, date: 11/20/2017, view: 8138, Q&A: 0
    [Abstract] Long-lived T-cell–mediated immunity requires persistence of memory T cells in an antigen-free environment while also maintaining a heightened capacity to recall effector functions. Such antigen-independent homeostatic proliferation is mediated in part by the common gamma-chain cytokines IL-7 and IL-15. To further explore the mechanisms governing ...
    ELISPOT Assay to Measure Peptide-specific IFN-γ Production
    Authors:  Michelle N. Wykes and Laurent Renia, date: 06/05/2017, view: 15758, 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, ...
    Intracellular Cytokine Staining (ICS) on Human Lymphocytes or Peripheral Blood Mononuclear Cells (PBMCs)
    Authors:  Sheena Gupta and Holden Maecker, date: 04/05/2015, view: 22258, Q&A: 1
    [Abstract] Production of cytokines plays an important role in the immune response. Cytokines are involved in many different pathways including the induction of many anti-viral proteins by IFN gamma, the induction of T cell proliferation by IL-2 and the inhibition of viral gene expression and replication by TNF alpha. Cytokines are not preformed factors but ...
    Intracellular Cytokine Staining on PBMCs Using CyTOFTM Mass Cytometry
    Authors:  Dongxia Lin, Sheena Gupta and Holden Maecker, date: 01/05/2015, view: 18858, Q&A: 2
    [Abstract] In this protocol, we use a CyTOFTM mass cytometry to collect single-cell data on a large number of cytokines/chemokines as well as cell-surface proteins that characterize T cells and other immune cells. The current selected mass window in AW 103-203 includes the lanthanides used for most antibody labeling, along with iridium and rhodium ...
    Measurement of TACE Activity in Extracts from Cultured Cells
    Authors:  Xiuling Xu, Mabel T. Padilla and Yong Lin, date: 10/20/2014, view: 7505, Q&A: 0
    [Abstract] In cigarette smoke–induced and inflammation-associated lung cancer development, cigarette smoke extract (CSE) activates tumor necrosis factor-alpha (TNF-α) secretion from macrophages. TNF-α converting enzyme (TACE), also known as α-Secretase or ADAM17 (A Disintegrin and Metalloprotease), is a member of the ADAM family of metalloproteases. TACE ...



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