Henry M. Jackson Foundation

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

A High Content Imaging Assay for Identification of Botulinum Neurotoxin Inhibitors

Krishna P. Kota^1,6, Veronica Soloveva^2,6,7, Laura M. Wanner^3,6, Glenn Gomba^4,6, Erkan Kiris^3,5,6, Rekha G. Panchal⁶, Christopher D. Kane^2,6,7, Sina Bavari⁶

¹Perkin Elmer Inc., ²Henry M. Jackson Foundation, ³The Geneva Foundation, ⁴ORISE, ⁵Frederick National Laboratory for Cancer Research, ⁶Division of Molecular and Translational Sciences, US Army Medical Research Institute of Infectious Diseases, ⁷DoD Biotechnology High Performance Computing Software Applications Institute (BHSAI), Telemedicine and Advanced Technology Research Center (TATRC), US Army Medical Research and Materiel Command (USAMRMC)

Botulinum neurotoxin is one of the most potent toxins among Category-A biothreat agents, yet a post-exposure therapeutic is not available. The high content imaging approach is a powerful methodology for identifying novel inhibitors as it enables multiparameter screening using biologically relevant motor neurons, the primary target of this toxin.

Immunology and Infection

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 Rolland^3,4, James I. Mullins^1,2

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.