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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: 2145, 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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Assessing in vitro and in vivo Trogocytosis By Murine CD4⁺ T cells

Authors: Jim Reed and Scott A. Wetzel, date: 05/05/2020, view: 2161, Q&A: 0

[Abstract] Recognition of antigens by lymphocytes (B, T, and NK) on the surface of an antigen-presenting cell (APC) leads to lymphocyte activation and the formation of an immunological synapse between the lymphocyte and the APC. At the immunological synapse APC membrane and associated membrane proteins can be transferred to the lymphocyte in a process called ...

In vitro Antigen-presentation Assay for Self- and Microbial-derived Antigens

Author: Laura Campisi, date: 06/05/2017, view: 13801, Q&A: 0

[Abstract] Antigen presenting cells (APC) are able to process and present to T cells antigens from different origins. This mechanism is highly regulated, in particular by Patter Recognition Receptor (PRR) signals. Here, I detail a protocol designed to assess in vitro the capacity of APC to present antigens derived from bacteria, apoptotic and ...

PKC-θ in vitro Kinase Activity Assay

Authors: Xudong Wang and Yingqiu Li, date: 10/20/2016, view: 7297, Q&A: 0

[Abstract] Protein kinase C-θ (PKC-θ), a member of the Ca²⁺-independent PKC subfamily of kinases, serves as a regulator of T cell activation by mediating the T cell antigen receptor (TCR)- and coreceptor CD28-induced activation of the transcription factors NF-κB and AP-1 and, to a lesser extent, NFAT, and, subsequently, interleukin 2 (IL-2) ...

Assessment of TCR-induced Sumoylation of PKC-θ

Authors: Xudong Wang, Zhilong Chen, Qilong Wang and Yingqiu Li, date: 10/20/2016, view: 5285, Q&A: 0

[Abstract] Sumoylation controls many cellular processes. Protein kinase C-θ (PKC-θ), a member of the Ca²⁺-independent PKC subfamily of kinases, serves as a regulator of T cell activation by mediating the T cell antigen receptor (TCR)- and coreceptor CD28-induced activation of the transcription factors NF-κB and AP-1 and, to a lesser extent, NFAT, ...

Peptide Loading on MHC Class I Molecules of Tumor Cells

Authors: Loredana Cifaldi, Franco Locatelli and Doriana Fruci, date: 09/20/2016, view: 8368, Q&A: 0

[Abstract] MHC class I molecules present peptides to cytotoxic T cells allowing the immune system to scan for intracellular pathogens and mutated proteins. The generation of antigenic peptides is a multistep process that ends in the endoplasmic reticulum (ER). Only peptides with the right length and sequence will bind nascent MHC class I molecules in the ER. ...

Killer Cell Ig-like Receptors (KIR)-Binding Assay for Tumor Cells

Authors: Loredana Cifaldi, Franco Locatelli and Doriana Fruci, date: 09/20/2016, view: 5225, Q&A: 0

[Abstract] Natural killer (NK) cells play key roles in innate and adaptive immune responses against virus and tumor cells. Their function relies on the dynamic balance between activating and inhibiting signals through receptors that bind ligands expressed on target cells. The absence of inhibitory receptor engagement with their ligands and the presence of ...

Efficient Isolation of Influenza Specific CTLs

Authors: Prerak Trivedi and Stuart I. Mannering, date: 06/20/2016, view: 6929, Q&A: 0

[Abstract] Human antigen-specific CD8⁺ T-cell clones are valuable tools for dissecting CD8⁺ T-cell responses against antigens derived from infectious agents, cancer and self antigens. Here we describe a protocol for isolating human antigen-specific CD8⁺ T cells. This protocol uses surface capture of IFNγ to identify antigen ...

In vivo OVA-specific Cytotoxic CD8⁺ T Cell Killing Assay

Authors: Nada Chaoul, Catherine Fayolle and Claude Leclerc, date: 06/20/2016, view: 15761, 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). ...

Adoptive Transfer of Memory B Cells

Authors: Griselda Zuccarino-Catania and Mark Shlomchik, date: 08/20/2015, view: 8497, 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 ...

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