Automated microarray platform for single‐cell sorting and collection of lymphocytes following <scp>HIV</scp> reactivation

A promising strategy to cure HIV-infected individuals is use latency reversing agents (LRAs) reactivate latent viruses, followed by host clearance of infected reservoir cells. However, reactivation proviruses within cells heterogeneous and often incomplete. This fact limits strategies HIV which may require complete elimination viable virus from all cellular reservoirs. For this reason, understanding the mechanism(s) reservoirs critical achieve therapeutic success. Methodologies enabling temporal tracking single as they sorting molecular analysis those are urgently needed. To end, microraft arrays were adapted image T-lymphocytes expressing mCherry under control long terminal repeat (LTR) promoter, in response application LRAs (prostratin, iBET151, SAHA). In prostratin, SAHA, 30.5%, 11.2%, 12.1% percentage cells, respectively. The enabled large numbers (>25,000) be imaged over time. fluorescence quantification identified cell subpopulations with differing kinetics. Significant heterogeneity was observed at single-cell level between different terms time reactivation, rate increase upon peak attained. T lymphocytes slow fast kinetics identified. Single that either or reactivators sorted, RNA-sequencing performed. Different genes associated inflammation, immune activation, viral transcription factors found.