Dual-color HIV reporters trace a population of latently infected cells and enable their purification.
HIV latency constitutes the main barrier for clearing HIV infection from patients. Our inability to recognize and isolate latently infected cells hinders the study of latent HIV. We engineered two HIV-based viral reporters expressing different fluorescent markers: one HIV promoter-dependent marker for productive HIV infection, and a second marker under a constitutive promoter independent of HIV promoter activity. Infection of cells with these viruses allows the identification and separation of latently infected cells from uninfected and productively infected cells. These reporters are sufficiently sensitive and robust for high-throughput screening to identify drugs that reactivate latent HIV. These reporters can be used in primary CD4 T lymphocytes and reveal a rare population of latently infected cells responsive to physiological stimuli. In summary, our HIV-1 reporters enable visualization and purification of latent-cell populations and open up new perspectives for studies of latent HIV infection.
Gladstone Institute of Virology and Immunology, 1650 Owens Street, San Francisco, CA 94158, USA; Department of Medicine, University of California, San Francisco, CA, USA.
Vincenzo
V
Calvanese
Leonard
L
Chavez
Timothy
T
Laurent
Sheng
S
Ding
Eric
E
Verdin
D015496:CD4-Positive T-Lymphocytes / Q000821:virology; D015658:HIV Infections / Q000473:pathology* / Q000821:virology*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology*; D006801:Humans; D013194:Staining and Labeling / Q000379:methods*; D014773:Virology / Q000379:methods*; D014775:Virus Activation / Q000187:drug effects; D017735:Virus Latency