3 articles published in JoVE
A High-throughput Shigella-specific Bactericidal Assay Hailey P. Weerts1, Jigui Yu2, Robert W. Kaminski1, Moon H. Nahm2 1Department of Enteric Infections, Bacterial Diseases Branch, Walter Reed Army Institute of Research, 2University of Alabama at Birmingham Here we present a protocol to measure Shigellacidal activity of antibodies in serum. Serum is mixed with bacteria and exogenous complement, incubated, and the reaction mixture is plated on agar plates. Viable bacteria form colonies which are counted, using an automated colony enumerator, and used to determine the bactericidal titer.
Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques Andrey Tokarev1, Matthew Creegan1, Michael A. Eller1, Mario Roederer2, Diane L. Bolton1 1US Military HIV Research Program, Henry M. Jackson Foundation, Walter Reed Army Institute of Research, 2Vaccine Research Center, NIAID, NIH Described is a methodology to quantitate the expression of 96 genes and 18 surface proteins by single cells ex vivo, allowing for the identification of differentially expressed genes and proteins in virus-infected cells relative to uninfected cells. We apply the approach to study SIV-infected CD4+ T cells isolated from rhesus macaques.
Pairwise Growth Competition Assay for Determining the Replication Fitness of Human Immunodeficiency Viruses Siriphan Manocheewa1, Erinn C. Lanxon-Cookson1, Yi Liu1, J. Victor Swain1, Jan McClure1, Ushnal Rao1, Brandon Maust1, Wenjie Deng1, Justine E. Sunshine1, Moon Kim1, Morgane Rolland3,4, James I. Mullins1,2 1Department of Microbiology, University of Washington, 2Departments of Medicine and Laboratory Medicine, University of Washington, 3U.S Military HIV Research Program, Walter Reed Army Institute of Research, 4Henry M. Jackson Foundation Growth competition between nearly isogenic viruses provides a sensitive measurement for determining relative replication fitness. The protocols described here include the construction of recombinant HIV-1 clones, virus propagation and growth competition and analysis methods optimized to yield sensitive and consistent results.