Core 3 - Immunobiology Core

Abstract – Core 3 – Immunobiology Core Approximately 40 million people worldwide are living with HIV/AIDS; however, a protective vaccine or functional cure remain elusive despite four decades of intense research. HIV-1 evades the immune system through its rapid structural evolution during infection and replication. The Duke Center for HIV Structural Biology will pursue structural studies of the evolution of the HIV-1 Envelope (Env) protein to elucidate structure-function mechanisms for viral entry, B-cell and T-cell activation, and viral rebound after antiretroviral therapy ART. The Immunobiology Core (Core 3) brings together a team of investigators with extensive expertise in cellular and molecular immunology to provide comprehensive, centralized immunoassay services to support the efforts of the three projects and the other cores. The overall objectives of the Immunobiology Core are to 1) support Projects 1, 2, and 3 with immunoassays and biochemical protocols to evaluate B cell receptor signaling in cell lines and knock- in mice, 2) provide assays for measurements of peptide presentation on MHCII as a read-out of B cell activation, and 3) support Project 3 by performing single B cell sorting from HIV-infected subjects for recovery and expression of Env-specific antibodies. The Immunobiology Core comprises facilities, technologies, and services housed in the cellular immunology laboratories of Dr. Cain (Core Lead) at the Duke Human Vaccine Institute (DHVI) and Dr. Borrow (Core Co-director) at Oxford University, the biomolecular interaction laboratory of Dr. Alam (Core Co-director) at DVHI, the protein production laboratory of Dr. Saunders (DHVI), and the DHVI Flow Cytometry Facility. In addition to interactions with individual projects, the Immunobiology Core will also directly interface with the Developmental Core. Core personnel will work hand-in-hand with trainees at the bench for the teaching of assays, with emphasis on robust experimental design, proper technique, and appropriate analysis of experimental results. As needed, the Core will host small-group workshops for trainees that focus on immunological topics or methodologies related to Core activities. The assays and services provided by the Immunobiology Core are critical to the success of the research projects. The provision of immunoassays will provide key insights into the propagation of signals induced by antigen-B cell receptor interactions at the B cell surface membrane, and the isolation of novel Env antibodies from HIV+ subjects will enable structural insights pertaining to suppression of the viral reservoir. Moreover, the methodologies and reagents developed in this Core will be made available to the research community for application in basic and applied structural immunology research.