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Marty and Helen Kimmel Center for Biology and Medicine and Skirball Institute for Biomolecular Medicine

2 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.

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.