Analytical treatment interruption: detection of an increase in the latent, inducible HIV-1 reservoir more than a decade after viral resuppression

Analytical treatment interruption (ATI) studies are increasingly being performed to evaluate the efficacy of putative strategies towards HIV-1 reservoir elimination or antiretroviral therapy (ART)-free viral control. A limited number of studies have evaluated the impact of ATI on the HIV-1 reservoir in individuals on suppressive ART. Available data suggests that ATIs have transient impact on the HIV-1 reservoir, mostly measured by levels of total or integrated HIV-1 DNA, in peripheral blood cells prior to ATI and shortly after ART-mediated viral re-suppression. The long-term impact of intervention ATI studies on the latent, inducible HIV-1 reservoir remains uncertain. We report the first clinical study demonstrating an increase in the latent, inducible HIV-1 reservoir, measured by expression of tat/rev multiply spliced RNA, in nine individuals, despite more than a decade of re-suppressive ART, after undergoing an immune intervention ATI conducted in 2006-2009. Our findings challenge the status quo on ATI risk of viral reservoir reseeding and the long-term outcomes thereof. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement C.L. received funding from the Dutch Aidsfonds (grants P-60602 and P-263); SR received funding from Dutch Aidsfonds (grant P-53302); C.R. received funding from Dutch Aidsfonds (grant P-53601); R.A.G. received funding from Dutch Aidsfonds (grant P-60804) and Horizon Europe (grant 681032);TMa received funding from Health Holland (grants LSHM19100-SGF and EMCLSH19023) and ZonMW (grant 40-44600-98-333) and a Building Synergistic Infrastructure for NL4Cure grant. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Ethics committee/IRB of Erasmus University Medical Center, Rotterdam gave ethical approval for this work (MEC-2005-227 and MEC-2012-583). I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data needed to evaluate the results and conclusions in this article are provided in the main text, Methods section and Extended data section. Additional data related to the study may be requested from the lead contact T. Mahmoudi. Unique biological material, where available, can be obtained upon request.