Detecting Sources of Immune Activation and Viral Rebound in HIV Infection
Stephen W. Wietgrefe, Lijie Duan, Jodi Anderson, Guillermo Marqués, Mark A Sanders, Nathan W. Cummins, Andrew D. Badley, Curtis Dobrowolski, Jonathan Karn, Amélie Pagliuzza, Nicolas Chomont, Gérémy Sannier, Mathieu Dubé, Daniel E. Kaufmann, Paul Zuck, Guoxin Wu, Bonnie J. Howell, Cavan Reilly, Alon Herschhorn, Timothy W. Schacker, Ashley T. Haase
Abstract
Anti-retroviral therapy (ART) has greatly improved the quality and length of life for people living with HIV, but immune activation does not normalize during ART, and persistent immune activation has been linked to increased morbidity and mortality. We report a comparison of assays of two potential sources of immune activation during ART: rare cells producing HIV and the virus' major viral protein, p24, benchmarked on a cell model of active and latent infections and a method to visualize HIV-producing cells. We show that assays of HIV envelope mRNA (EDITS assay), gag mRNA, and p24 (Flow-FISH, HIV-Flow. and ultrasensitive p24 immunoassay) detect HIV-producing cells and p24 at sensitivities of one infected cell in a million uninfected cells, thereby providing validated tools to explore sources of immune activation during ART in the lymphoid and other tissue reservoirs.