Immunology

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    Protocols in Current Issue
    Optogenetic Tuning of Ligand Binding to The Human T cell Receptor Using The opto-ligand-TCR System
    [Abstract] T cells are one major cell type of the immune system that use their T cell antigen receptor (TCR) to bind and respond to foreign molecules derived from pathogens. The ligand-TCR interaction half-lives determine stimulation outcome. Until recently, scientists relied on mutating either the TCR or its ligands to investigate how varying TCR-ligand ...
    Quantifying HIV-1-Mediated Gut CD4+ T Cell Death in the Lamina Propria Aggregate Culture (LPAC) Model
    [Abstract] Gut CD4 T cells are major targets of HIV-1 and are massively depleted early during infection. To better understand the mechanisms governing HIV-1-mediated CD4 T cell death, we developed the physiologically-relevant Lamina Propria Aggregate Culture (LPAC) model. The LPAC model is ideal for studying CD4 T cell death induced by clinically-relevant ...
    Measurement of CD74 N-terminal Fragment Accumulation in Cells Treated with SPPL2a Inhibitor
    Authors:  Rubén Martínez-Barricarte, Xiao-Fei Kong and Jean-Laurent Casanova, date: 06/05/2019, view: 1229, Q&A: 0
    [Abstract] The recent discovery of human signal peptide peptidase-like 2a (SPPL2a) deficiency in humans revealed the toxicity associated with the accumulation of one of its substrates, CD74 N-terminal fragment (CD74-NTF), for certain type of dendritic cells (cDC2). We developed a two-step protocol for monitoring the accumulation of this molecule in different ...
    Induced Germinal Center B Cell Culture System
    Authors:  Kei Haniuda and Daisuke Kitamura, date: 02/20/2019, view: 4049, 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 ...
    Assessment of Humoral Alloimmunity in Mixed Lymphocyte Reaction
    Authors:  Georgios Pissas and Theodoros Eleftheriadis, date: 01/20/2019, view: 3128, Q&A: 1
    [Abstract] Humoral alloimmunity remains a significant and unresolved problem that constrains allograft survival. Thus, there is a need for the development of an easy, preferably non-radioactive, and inexpensive protocol for assessing the effect of various drug treatments on humoral alloimmunity. In order to satisfy this demand, we developed such a protocol ...
    Integrin Dependent RhoB Activation Assay Using Leukocytes
    Authors:  Janne Lehtonen, Katarzyna Potrzebowska, Malin Samuelsson and Lena Svensson, date: 12/05/2018, view: 1074, Q&A: 0
    [Abstract] To be able to migrate, leukocyte needs to re-use its adhesion molecules to move forward. These adhesion molecules are called integrins and are intracellularly transported via endocytosis and exocytosis in order to translocate to a new site on the cell membrane. The intracellular transportation is regulated by different small GTPases including ...
    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: 10233, 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 vitro Assessment of Immunological Synapse Formation by Flow Cytometry
    Authors:  Bo-Ra Na and Chang-Duk Jun, date: 03/20/2016, view: 11074, Q&A: 2
    [Abstract] In adaptive immune system, formation of immunological synapse between T cells and antigen presenting cells (dendritic cells, B cells, and macrophages) or target cells (tumor cells and viral-infected cells) is critical for the execution of T cell immune responses via cytokine secretion or direct killing activity. Here, we describe the practical ...
    Mouse BMDC-dependent T Cell Polarization Assays
    Author:  Vijaya Satchidanandam, date: 02/05/2016, view: 9119, Q&A: 0
    [Abstract] In response to exposure to antigen, T cells whose T cell receptor (TCR) are capable of recognizing the self MHC-antigen derived peptide complex, respond to the antigen and differentiate into one of several subsets, namely TH1, TH2, TH17, Treg, etc. characterized by the signature cytokine they secrete, namely IFN-γ, IL-4, IL-17 or IL-10, ...
    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: 20541, 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 ...



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