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The University of Oxford

4 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Imaging of HIV-1 Envelope-induced Virological Synapse and Signaling on Synthetic Lipid Bilayers

Kathleen C. Prins^*1,2, Gaia Vasiliver-Shamis^*2,3, Michael Cammer^1,2, David Depoil^1,2, Michael L. Dustin², Catarina E. Hioe^1,4

¹Department of Pathology, New York University Langone School of Medicine, ²Program in Molecular Pathogenesis, Marty and Helen Kimmel Center for Biology and Medicine and Skirball Institute for Biomolecular Medicine, ³Laboratory of Molecular Immunogenetics, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, ⁴Veteran Affairs New York Harbor Healthcare System

This article describes a method to visualize formation of an HIV-1 envelope-induced virological synapse on glass supported planar bilayers by total internal reflection fluorescence (TIRF) microscopy. The method can also be combined with immunofluorescence staining to detect activation and redistribution of signaling molecules that occur during HIV-1 envelope-induced virological synapse formation.

Medicine

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

Chrystalina A. Antoniades¹, James J. FitzGerald²

¹Nuffield Department of Clinical Neuroscience, The University of Oxford, ²Nuffield Department of Surgical Science, The University of Oxford

This paper describes the use of quantitative measurement of eye movements in conjunction with stimulation of focal areas of the deep brain in order to study physiology, pathophysiology, and the mechanisms of deep brain stimulation.

Immunology and Infection

Imaging of In Situ Interferon Gamma Production in the Mouse Spleen following Listeria monocytogenes Infection

Julie M. Mazet¹, Ana L. Chiodetti¹, Jagdish N. Mahale¹, Audrey Gérard¹

¹The Kennedy Institute of Rheumatology, The University of Oxford

Here, we describe a simple confocal imaging method to visualize the in situ localization of cells secreting the cytokine Interferon gamma in murine secondary lymphoid organs. This protocol can be extended for the visualization of other cytokines in diverse tissues.

Immunology and Infection

Preparation of Bead-supported Lipid Bilayers to Study the Particulate Output of T Cell Immune Synapses

Pablo F. Céspedes¹, Michael L. Dustin¹

¹Kennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, The University of Oxford

Here we present the protocol for the stepwise reconstitution of synthetic antigen-presenting cells using Bead-Supported Lipid Bilayers and their use to interrogate the synaptic output from activated T cells.