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The \u201cshock and kill\u201d strategy aims to eliminate latent reservoirs by inducing viral transcription through latency-reversing agents (LRAs), thereby exposing infected cells to immune-mediated clearance. We developed and validated a simple, low-cost, and highly reproducible\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      screening protocol to evaluate the efficacy and safety of LRAs, using a two-color flow cytometry assay on ACH2 cells, a well-characterized model of HIV-1 latency. Using this method, we identified PEP005 and CUDC-907 as the most potent LRAs across multiple experimental settings. Their reactivation capacity was further confirmed through transcriptomic analysis, which revealed a significant upregulation of viral RNA copies following stimulation. In addition, we collaborated with Domp\u00e9 for using the Exscalate platform, an innovative computer-aided drug discovery approach, together with the experimental validation, which led to the identification of Tandutinib, a tyrosine kinase and mTOR inhibitor, as a novel LRA candidate with appreciable latency-reversing activity in the ACH2 model. While the ACH2 cell line does not fully recapitulate the complexity of HIV latency\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      , it offers a robust and scalable system for early-stage screening and prioritization of candidate LRAs. Importantly, the future application of these LRAs in\n                      <jats:italic toggle=\"yes\">ex vivo<\/jats:italic>\n                      samples derived from people living with HIV, with a particular focus on pediatric samples, will be crucial to deepen our understanding of latency reactivation in clinically relevant settings and age-specific immune environments.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>This study addresses a major obstacle to curing human immunodeficiency virus type 1 (HIV-1) infection: the persistence of latent viral reservoirs that are not eliminated by current therapies. We developed a simple and reproducible assay to identify compounds capable of reactivating latent virus, a key step in cure strategies. Using this approach, we identified effective latency-reversing compounds and, through collaboration with Domp\u00e9 using the EXSCALATE platform, we also identified Tandutinib as a promising new candidate for further investigation.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.00407-26","type":"journal-article","created":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T13:00:36Z","timestamp":1785848436000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Selection of effective LRAs using a newly designed\n                    <i>in vitro<\/i>\n                    HIV latency reactivation protocol: toward future application in HIV 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