Abstract Despite suppressive antiretroviral therapy (ART), HIV persists in the central nervous system (CNS) and contributes to HIV-associated neurocognitive disorder (HAND), but cell type–specific effects remain poorly defined. Using fluorescence-activated nuclei sorting of postmortem brain tissue of aviremic and viremic deceased people with HIV (DPWH) and HIV-negative individuals, we quantified the size of the HIV CNS reservoir and transcriptional alterations. Microglia were identified as the dominant CNS reservoir, harboring 10³–10⁴ HIV DNA copies per million cells by ddPCR-LTR assay in both aviremic and viremic DPWH. Bulk RNA-sequencing revealed immune pathway upregulation specifically in microglia, and downregulation of synaptic and homeostatic pathways across cell-types in viremic compared to aviremic individuals. ART partially mitigated microglial transcriptional dysregulation, but transcriptional profiles did not restore profiles to HIV-negative levels. Notably, persistent microglial infection was associated with transcriptional changes in other cell-types, underscoring microglia as a key therapeutical target for CNS-directed HIV cure strategies.

IntroductionDespite effective antiretroviral therapy, HIV-1 persists as a viral reservoir and remains a major barrier to a durable cure. Reservoir size and dynamics are central determinants of the success of cure strategies. Efforts to characterize and target the HIV-1 reservoir have primarily focused on people with HIV-1 subtype B in Western Europe and North America. This emphasis has limited our understanding of reservoir size and dynamics in individuals with non-B subtypes, which account for most infections globally, particularly in high-burden regions such as sub-Saharan Africa. This systematic review synthesizes evidence on HIV-1 reservoir size in individuals with non-B subtypes and compiles reported reservoir outcomes alongside demographic and clinical metadata.MethodsPubMed and EMBASE were searched for studies quantifying the HIV-1 reservoir in peripheral blood from individuals with non-B subtypes. Eligible studies quantified reservoir size in at least three non-B samples stratified by subtype or directly compared measurements between subtypes. Data were extracted on subtype, reservoir size, statistical comparisons thereof, and key demographic and clinical data. Study quality was assessed using a pre-defined, custom set of criteria. Data were synthesized by organizing within-study statistical comparisons by subtype in a schematic overview to indicate the direction of effect.ResultsOf 1589 unique records, 40 studies quantifying HIV-1 reservoir size in individuals with non-B subtypes were included. Records excluded at full-text screening commonly lacked viral subtype determination and/or subtype-stratified outcomes. Comparisons between subtype B and pooled non-B subtypes suggest a larger reservoir for subtype B. Substantial differences in study design, (meta)data reporting, and comparability limited direct comparison across studies.ConclusionStrengthening knowledge on the HIV-1 subtype-specific reservoir is essential for the development of effective and globally relevant cure strategies. The available data point toward possible differences in reservoir size between subtype B and non-B subtypes. To improve further assessment, we developed recommendations urging studies to use standardized subtyping methods, report viral subtype according to Los Alamos guidelines, and provide detailed individual-level outcomes and metadata, including disease stage at treatment initiation, ART duration, and viral suppression status. Furthermore, we strongly recommend using multi-subtype reservoir assays that are cross-validated to enhance comparability across studies.Systematic Review Registrationhttps://www.crd.york.ac.uk/PROSPERO/view/CRD420251074799, identifier CRD420251074799.

Despite the success of antiretroviral therapy, a subset of people with HIV experience low-level viraemia (LLV), a challenging condition with inconsistent definitions and management across settings. We conducted a scoping review of the literature and developed international consensus recommendations through a modified Delphi process involving a multidisciplinary panel of experts. Available evidence shows wide variability in definitions of persistent LLV, most commonly encompassing repeated viral loads between 50 and 1000 copies per mL. Outcomes associated with LLV are heterogeneous. Despite associations between LLV at 50-200 copies per mL and virological failure of more than 1000 copies per mL, resistance emergence or adverse clinical outcomes are inconsistent; stronger evidence links LLV of 200-1000 copies per mL to these endpoints. Based on these findings, we propose a pragmatic management framework that includes prompt confirmation of LLV, assessment of adherence and drug interactions, consideration of resistance testing (especially at viral loads of 200-1000 copies per mL), a broader clinical evaluation, individualised treatment optimisation (according to viral load strata, the genetic barrier to resistance of current and potential antiretroviral regimens, and the HIV resistance profile), and individual prevention strategies for viral loads of 200-1000 copies per mL. This international guidance aims to support clinicians in identifying when LLV requires action, reduce variability in clinical practice, and inform management strategies across diverse health-care settings.

BACKGROUND:Dolutegravir resistance is increasingly reported in treatment-experienced people with HIV in low-income and middle-income countries. Resistance testing resources are sparse, and viral rebound often reflects adherence lapses rather than resistance. Efficient strategies are needed to target testing to individuals at highest risk. We evaluated a reflex laboratory approach using plasma dolutegravir exposure to guide resistance testing in routine care. METHODS:In this prospective implementation study at two public-sector clinics in Johannesburg, South Africa, we analysed adults (aged ≥18 years) on dolutegravir-based antiretroviral therapy (ART) with viral rebound of 400 copies per mL or higher. From May 18, 2023, to Jan 10, 2025, we tested residual routine viral load samples for dolutegravir exposure using an enzyme immunoassay. If dolutegravir was detectable (≥20 ng/mL), we did reflex resistance testing on the same specimen; retrospective sequencing was done at study end for dolutegravir-undetectable samples. We modelled costs comparing the reflex approach with standard of care. FINDINGS:We analysed 400 samples from 288 individuals; 160 (56%) were female and 128 (44%) male, the median age was 44 years (IQR 35-50), and 240 (83%) had previous non-dolutegravir ART exposure. Plasma dolutegravir was detectable in 108 (38%) participants. Among those with successful sequencing, dolutegravir resistance occurred in 13 (13%) of 102 participants with detectable dolutegravir and three (2%) of 138 with undetectable dolutegravir, yielding an overall prevalence of 6·7% dolutegravir resistance and a negative predictive value for absence of dolutegravir resistance of 97·8% (95% CI 93·5-99·5). Dolutegravir resistance was associated with detectable dolutegravir (odds ratio [OR] 3 [95% CI 1-9], p=0·031), nucleoside reverse transcriptase inhibitor mutations (OR 33 [4-264], p=0·0009), non-nucleoside reverse transcriptase inhibitor mutations (OR 17 [2-135], p=0·0068), or M184V (OR 20 [4-92], p=0·0002). Cost modelling showed a 23% reduction in monitoring cost per person using the reflex strategy after confirmed viraemia versus current national guidelines. INTERPRETATION:Laboratory-initiated reflex plasma dolutegravir exposure testing provides an objective, cost-efficient strategy to predict the absence of dolutegravir resistance and supports adherence management, enabling timely, individualised treatment decisions. Reflex dolutegravir exposure testing could enhance clinical decision making, improve treatment outcomes, and strengthen HIV treatment and resistance monitoring algorithms in programmatic settings. FUNDING:The Amsterdam Dinner Foundation and Aidsfonds, the Netherlands Organisation for Scientific Research, ZonMw, and WOTRO.

A proviral reservoir persists within the central nervous system (CNS) of people with HIV, but its characteristics remain poorly understood. Research has primarily focused on cerebrospinal fluid (CSF), as acquiring brain tissue is challenging. We examined size, cellular tropism, and infection-dynamics of the viral reservoir in post-mortem brain tissue from five individuals on and off antiretroviral therapy (ART) across three brain regions. Microglia-enriched fractions (CD11b+) were isolated and levels of intact proviral DNA were quantified (IPDA). Full-length envelope reporter viruses were generated and characterized in CD4+ T cells and monocyte-derived microglia. HIV DNA was observed in microglia-enriched fractions of all individuals, but intact proviruses were identified only in one ART-treated individual, representing 15% of the total proviruses. Phenotypic analyses of clones from this individual showed that 80% replicated efficiently in microglia and CD4+ T cells, while the remaining viruses replicated only in CD4+ T cells. No region-specific effects were observed. These results indicate a distinct HIV brain reservoir in microglia for all individuals, although intact proviruses were detected in only one. Given the unique immune environment of the CNS, the characteristics of microglia, and the challenges associated with targeting these cells, the CNS reservoir should be considered in cure strategies.

Objectives: To investigate the short- and long-term dynamics of intact and defective proviral HIV DNA during ART. Design: We evaluated viral reservoir dynamics in a cohort of nine individuals with chronic HIV-1 subtype B infection who initiated first-line ART and were followed for 20 years while continuing ART. Methods: PBMCs were obtained before ART (n = 5), during the first year, and after 8.5 and 20 years of treatment. T cell subsets (naive, central-memory, transitional-memory and effector-memory) were sorted at 8.5 and 20 years. DNA was isolated and analyzed using the intact proviral DNA assay (IPDA). Deep-sequencing of the viral env region enabled analysis of viral evolution and cellular mechanisms underlying HIV persistence. Results: Initially, defective and intact proviral DNA in PBMCs declined with half-lives of 3.6 and 5.4 weeks, respectively. Over the following 8.5 years, the intact reservoir continued to decrease, with a half-life of 18.8 months in PBMCs, while defective proviral DNA levels stabilized. After 8.5 and 20 years of ART, the intact reservoir showed no further decline, with most intact proviral DNA residing in memory T cell subsets. Phylogenetic analysis revealed no signs of viral evolution over time, both within and between T cell subsets. Conclusions: PBMCs containing intact proviral DNA are selectively lost during the first decade of suppressive ART, followed by a decade of stabilization of this reservoir in the memory T cell subsets. In the absence of clear signs of viral evolution and massive clonal expansion, homeostatic proliferation might be an important driver of HIV persistence during long-term ART.

Background Real-world data showing the long-term effectiveness of long-acting injectable cabotegravir and rilpivirine are scarce. We assessed the effectiveness of cabotegravir and rilpivirine in all individuals who switched to cabotegravir and rilpivirine in the Netherlands. Methods We used data from the ATHENA cohort, an ongoing observational nationwide HIV cohort in the Netherlands. In the primary analysis, we matched individuals who commenced cabotegravir and rilpivirine and had no history of virological failure (ie, one or more measurements of a plasma HIV RNA >= 1000 copies per mL; hereafter referred to as exposed) 1:2 with individuals using oral antiretroviral therapy (ART; hereafter referred to as unexposed). We assessed the effectiveness of cabotegravir and rilpivirine using restricted mean survival time (RMST) until loss of virological control (one or more measurements of plasma HIV RNA >= 200 copies per mL). In the secondary analysis, we assessed loss of virological control in individuals who commenced cabotegravir and rilpivirine with previous virological failure or unsuppressed HIV-1 RNA at cabotegravir and rilpivirine initiation, or both. Findings In primary analysis, 585 exposed and 1170 unexposed individuals were included between Feb 27, 2018, and Aug 17, 2023. Median follow-up was 13 years (IQR 09 to 17). 14 exposed (2%) and 29 unexposed (2%) individuals had a loss of virological control, with no difference in RMST (difference=0026, 95% CI -0029 to -0080). Seven (50%) exposed individuals re-suppressed without a regimen change. Seven (50%) switched ART, and six (43%) of 14 had documented integrase strand transfer inhibitor (INSTI) or non-nucleoside reverse transcriptase inhibitor (NNRTI) resistance. No unexposed individuals switched ART after loss of virological control. In the secondary analysis, 105 individuals were included between July 1, 2016, and Aug 17, 2023. During a median follow up of 14 years (IQR 08 to 18), nine (9%) had a loss of virological control, of which five (56%) had INSTI or NNRTI resistance. Interpretation Switching to cabotegravir and rilpivirine was not associated with a higher risk of loss of virological control among individuals without previous virological failure compared with oral ART. The high risk of loss of virological control among individuals with previous virological failure or an unsuppressed HIV-1 RNA at cabotegravir and rilpivirine initiation warrants more careful monitoring. Copyright (c) 2025 Elsevier Ltd. All rights reserved, including those for text and data mining, AI training, and similar technologies.

Human immunodeficiency virus type 2 (HIV-2) is an attenuated retroviral infection characterized by specific natural susceptibility to antiretroviral drugs and acquired resistance profiles. Based on the latest knowledge of phenotypic data and clinical follow-up, HIV-2 resistance interpretation rules have been updated and implemented in a freely available resistance analysis tool.

BackgroundAllogeneic haematopoietic stem-cell transplantation (allo-HSCT) markedly reduces HIV reservoirs, but the mechanisms by which this occurs are only partly understood. In this study, we aimed to describe the dynamics of virological and immunological markers of HIV persistence after allo-HSCT.MethodsIn this prospective observational cohort study, we analysed the viral reservoir and serological dynamics in IciStem cohort participants with HIV who had undergone allo-HSCT and were receiving antiretroviral therapy, ten of whom had received cells from donors with the CCR5Δ32 mutation. Participants from Belgium, Canada, Germany, Italy, the Netherlands, Spain, Switzerland, and the UK were included in the cohort both prospectively and retrospectively between June 1, 2014 and April 30, 2019. In the first 6 months after allo-HSCT, participants had monthly assessments, with annual assessments thereafter, with the protocol tailored to accommodate for the individual health status of each participant. HIV reservoirs were measured in blood and tissues and HIV-specific antibodies were measured in plasma. We used the Wilcoxon signed-rank test to compare data collected before and after allo-HSCT in participants for whom longitudinal data were available. When the paired test was not possible, we used the Mann-Whitney U test. We developed a mathematical model to study the factors influencing HIV reservoir reduction in people with HIV after allo-HSCT.FindingsWe included 30 people with HIV with haematological malignancies who received a transplant between Sept 1, 2009 and April 30, 2019 and were enrolled within the IciStem cohort and included in this analysis. HIV reservoirs in peripheral blood were reduced immediately after full donor chimerism was achieved, generally accompanied by undetectable HIV-DNA in bone marrow, ileum, lymph nodes, and cerebrospinal fluid, regardless of donor CCR5 genotype. HIV-specific antibody levels and functionality values declined more slowly than direct HIV reservoir values, decaying significantly only months after full donor chimerism. Mathematical modelling suggests that allogeneic immunity mediated by donor cells is the main viral reservoir depletion mechanism after massive reservoir reduction during conditioning chemotherapy before allo-HSCT (half-life of latently infected replication-competent cells decreased from 44 months to 1·5 months).InterpretationOur work provides, for the first time, data on the effects of allo-HSCT in the context of HIV infection. Additionally, we raise the question of which marker can serve as the last reporter of the residual viraemia, postulating that the absence of T-cell immune responses might be a more reliable marker than antibody decline after allo-HSCT.FundingamfAR (American Foundation for AIDS Research; ARCHE Program), National Institutes of Health, National Institute of Allergy and Infectious Diseases, and Dutch Aidsfonds.

There is an ongoing debate regarding whether low-level viremia (LLV), in particular persistent LLV, during HIV treatment with optimal adherence originates from low-level viral replication, viral production, or both. We performed an observational study in 30 individuals with LLV who switched to a boosted darunavir (DRV)-based therapy. In-depth virological analyses were used to characterize the viral population and the (activity) of the viral reservoir. Immune activation was examined using cell-bound and soluble markers. The primary outcome was defined as the effect on HIV-RNA and was categorized by responders (<50 cp/mL) or non-responders (>50 cp/mL). At week 24, 53% of the individuals were considered responders, 40% non-responders, and 7% could not be assigned. Sequencing showed no evolution or selection of drug resistance in the non-responders. Production of defective virus with mutations in either the protease (D25N) or RT active site contributed to persistent LLV in two individuals. We show that in about half of the study participants, the switch to a DRV-based regimen resulted in a viral response indicative of ongoing low-level viral replication as the cause of LLV before the switch. Our data confirm that in clinical management, high genetic barrier drugs like DRV are a safe choice, irrespective of the source of LLV.

Journal Article Corrected proof Reply to Buzón-Martín and Troya Get access Berend J Van Welzen, Berend J Van Welzen Department of Infectious Diseases, University Medical Center Utrecht, The Netherlands Correspondence: B. J. van Welzen, Department of Infectious Diseases, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands (B.J.vanWelzen@umcutrecht.nl). https://orcid.org/0000-0001-5601-4207 Search for other works by this author on: Oxford Academic PubMed Google Scholar David Burger, David Burger Department of Pharmacy, Institute of Medical Innovation, Radboud University Medical Center, Nijmegen, The Netherlands Search for other works by this author on: Oxford Academic PubMed Google Scholar Annemarie M J Wensing Annemarie M J Wensing Translational Virology, Department of Medical Microbiology, University Medical Center Utrecht, The NetherlandsEzintsha, Department of Health, University of the Witwatersrand, Johannesburg, South Africa Search for other works by this author on: Oxford Academic PubMed Google Scholar Clinical Infectious Diseases, ciae142, https://doi.org/10.1093/cid/ciae142 Published: 14 March 2024 Article history Published: 14 March 2024 Corrected and typeset: 06 April 2024

A novel host-protein score (called MMBV) helps to distinguish bacterial from viral infection by combining the blood concentrations of three biomarkers: tumour necrosis factor related apoptosis inducing ligand (TRAIL), interferon gamma induced protein 10 (IP-10), and C-reactive protein (CRP). These host biomarkers are differentially expressed in response to bacterial versus viral acute infection. We conducted a prospective study, with a time series design, in healthy adult volunteers in the Netherlands. The aim was to determine the variability of TRAIL, IP-10, and CRP and the MMBV score in healthy adults across time. Up to six blood samples were taken from each healthy volunteer over a period of up to four weeks. In 77 healthy participants without recent or current symptoms, MMBV scores (maximal) were bacterial in 1.3 % and viral (or other non-infectious etiology) in 93.5 % of participants. There was little variation in the mean concentrations of TRAIL (74.5 pg/ml), IP-10 (113.6 pg/ml), and CRP (1.90 mg/L) as well as the MMBV score. The variability of biomarker measurement was comparable to the precision of the measurement platform for TRAIL, IP-10, and CRP. Our findings establish the mean values of these biomarkers and MMBV in healthy individuals and indicate little variability between and within individuals over time, supporting the potential utility of this novel diagnostic to detect infection-induced changes.

Introduction HIV reservoir quantification is essential for evaluation of HIV curative strategies and may provide valuable insights about reservoir dynamics during antiretroviral therapy. The Intact Proviral DNA Assay (IPDA) provides the unique opportunity to quantify the intact and defective reservoir. The current IPDA is optimized for HIV-1 subtype B, the dominant subtype in resource-rich settings. However, subtype C is dominant in Sub-Saharan Africa, jointly accounting for around 60% of the pandemic. We developed an assay capable of quantifying intact and defective proviral HIV-1 DNA of subtype B and C. Methods Primer and probe sequences were strategically positioned at conserved regions in psi and env and adapted to subtype B&C. In silico analysis of 752 subtype B and 697 subtype C near-full length genome sequences (nFGS) was performed to predict the specificity and sensitivity. Gblocks were used to determine the limit of blank (LoB), limit of detection (LoD), and different annealing temperatures were tested to address impact of sequence variability. Results The in silico analysis showed that the HIV-1 B&C IPDA correctly identified 100% of the intact subtype B, and 86% of the subtype C sequences. In contrast, the original IPDA identified 86% and 12% of these subtype B and C sequences as intact. Furthermore, the HIV-1 B&C IPDA correctly identified hypermutated (87% and 88%) and other defective sequences (73% and 66%) for subtype B and C with comparable specificity as the original IPDA for subtype B (59% and 63%). Subtype B cis-acting sequences were more frequently identified as intact by the HIV-1 B&C IPDA compared to the original IPDA (39% and 2%). The LoB for intact proviral DNA copies was 0, and the LoD for intact proviral DNA copies was 6 (> 95% certainty) at 60 °C. Quantification of 2–6 copies can be performed with > 80% certainty. Lowering the annealing temperature to 55 °C slightly lowered the specificity but prevented exclusion of samples with single mutations in the primer/probe region. Conclusions We developed a robust and sensitive assay for the quantification of intact and defective HIV-1 subtype B and C proviral DNA, making this a suitable tool to monitor the impact of (large-scale) curative interventions.

HIV cure has been reported for five individuals who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) with cells from CCR5 Delta 32 homozygous donors. By contrast, viral rebound has occurred in other people living with HIV who interrupted antiretroviral treatment after undergoing allo-HSCT, with cells mostly from wild-type CCR5 donors. Here we report the case of a male individual who has achieved durable HIV remission following allo-HSCT with cells from an unrelated HLA-matched (9 of 10 matching for HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 alleles) wild-type CCR5 donor to treat an extramedullary myeloid tumor. To date, plasma viral load has remained undetectable for 32 months after the interruption of antiretroviral treatment. Treatment with ruxolitinib has been maintained during this period to treat chronic graft-versus-host disease. Low levels of proviral DNA were detected sporadically after allo-HSCT, including defective but not intact HIV DNA. No virus could be amplified in cultures of CD4+ T cells obtained after antiretroviral treatment interruption, while CD4+ T cells remained susceptible to HIV-1 infection in vitro. Declines in HIV antibodies and undetectable HIV-specific T cell responses further corroborate the absence of viral rebound after antiretroviral treatment interruption. These results suggest that HIV remission could be achieved in the context of allo-HSCT with wild-type CCR5. HIV remission was achieved for 32 months after allogeneic hematopoietic stem cell transplantation with wild-type CCR5 donor cells.

IntroductionEvidence-based intervention strategies to improve adherence among individuals living with chronic conditions are critical in ensuring better outcomes. In this systematic review, we assessed the impact of interventions that aimed to promote adherence to treatment for chronic conditions.MethodsWe systematically searched PubMed, Web of Science, Scopus, Google Scholar and CINAHL databases to identify relevant studies published between the years 2000 and 2023 and used the QUIPS assessment tool to assess the quality and risk of bias of each study. We extracted data from eligible studies for study characteristics and description of interventions for the study populations of interest.ResultsOf the 32,698 total studies/records screened, 2814 were eligible for abstract screening and of those, 497 were eligible for full-text screening. A total of 82 studies were subsequently included, describing a total of 58,043 patients. Of the total included studies, 58 (70.7%) were related to antiretroviral therapy for HIV, 6 (7.3%) were anti-hypertensive medication-related, 12 (14.6%) were anti-diabetic medication-related and 6 (7.3%) focused on medication for more than one condition. A total of 54/82 (65.9%) reported improved adherence based on the described study outcomes, 13/82 (15.9%) did not have clear results or defined outcomes, while 15/82 (18.3%) reported no significant difference between studied groups. The 82 publications described 98 unique interventions (some studies described more than one intervention). Among these intervention strategies, 13 (13.3%) were multifaceted (4/13 [30.8%] multi-component health services- and community-based programmes, 6/13 [46.2%] included individual plus group counselling and 3/13 [23.1%] included SMS or alarm reminders plus individual counselling).DiscussionThe interventions described in this review ranged from adherence counselling to more complex interventions such as mobile health (mhealth) interventions. Combined interventions comprised of different components may be more effective than using a single component in isolation. However, the complexity involved in designing and implementing combined interventions often complicates the practicalities of such interventions.ConclusionsThere is substantial evidence that community- and home-based interventions, digital health interventions and adherence counselling interventions can improve adherence to medication for chronic conditions. Future research should answer if existing interventions can be used to develop less complicated multifaceted adherence intervention strategies.

HIV can be successfully suppressed to undetectable levels by antiretroviral therapy (ART) in most people with HIV (PWH). However, a small proportion continues to have persistent low-level viremia (LLV) during ART. A presumed source of LLV is production or replication from viral reservoirs, which are maintained in the presence of ART. It is unknown whether the oral cavity can be considered an HIV reservoir. As periodontal inflammation is a common problem in PWH, we hypothesize that periodontal inflammation in the oral cavity activates (latently) infected cells and thus might be associated with LLV. We included 11 individuals with HIV LLV, and compared HIV-RNA levels in saliva and plasma at baseline and at week 24 after switch of ART. We compared the LLV-group at baseline with 11 age-matched controls with suppressed viremia. To investigate the severity of periodontitis we used Periodontal Inflamed Surface Areas (PISA) by measuring probing depth, gingival recession, bleeding on probing and clinical attachment level. Severity of periodontitis was classified according to the CDC-AAP case definition. Additional insights in periodontal inflammation were obtained by comparing immune activation markers and the presence of periodontal pathogens. In four individuals of the LLV group, residual levels of HIV-RNA were detected in saliva at baseline (N = 1) or at week 24 (N = 2) or both (N = 1). Of the four individuals with LLV, three had residual levels of HIV-RNA in saliva. All 22 individuals had moderate to severe periodontitis. PISA was not significantly different between cases with LLV and controls. Similarly, periodontal pathogens were frequently observed in both groups. Total activated HLA-DR+CD38+ CD4+ cells and CD8+ cells were significantly higher in the LLV group than in the control group (p = <0.01). No immune markers were associated with LLV. In conclusion, periodontal inflammation is an unlikely driver of HIV LLV compared to HIV suppressed individuals.

IntroductionThe main obstacle to achieving an HIV-1 cure is the proviral reservoir. To promote equity in HIV cure strategies, it is crucial to study the viral reservoir of the predominant HIV-1 subtype C in both women and men. Therefore, we investigated the dynamics of the (intact) viral reservoir in relation to plasma viral load (VL), CD4+ T cell count, and immune activation before and during 96 weeks of successful antiretroviral therapy (ART).MethodsEighty-two participants (62% female) newly initiating ART in a rural clinic in South Africa were included in the study. Blood samples were collected at baseline, week 48, and week 96, and CD4 count was determined. Plasma was used for VL and immune marker analyses, while isolated peripheral blood mononuclear cells (PBMCs) were used for the quantification of cellular multiple spliced HIV-1 RNA (msRNA) and the intact proviral DNA assay. For the longitudinal analyses on ART, we selected only those participants who durably suppressed their VL to <200 copies/mL during 48 (n=65) and/or 96 (n=60) weeks of treatment.ResultsAt ART initiation, the median CD4 count was 234 cells/mm3 and VL was 68,897 copies/mL. Interestingly, at baseline the number of defective proviruses was significantly correlated with VL (p<0.0001), msRNA (p<0.0001), CD4 count (p=0.0008), CXCL10 (p=0.0003) and TNF-α (p=0.0394). During successful ART, a significant decrease of both the intact and defective proviral reservoir was observed (p<0.0001). The decrease of the intact proviral reservoir was more profound compared to the defective fraction after 96 weeks of therapy. In addition, a significant decrease in cellular msRNA and IL-6, IL-7, TNF-α, sCD14, sCD163, CCL2, CXCL10, and CRP was detected.DiscussionThis study underscores the significant relationship observed prior to therapy initiation between the number of defective proviruses, viral transcription/production and their association with immune response indicators such as CD4 count, CXCL10, and TNF-α. Furthermore, the observation of a less pronounced decrease of the defective proviral DNA highlights the importance of addressing both intact and defective proviruses in therapeutic strategies to enhance clinical outcomes for people with HIV-1. Together, these findings suggest a significant role of the defective proviruses in HIV-related disease progression.

Background Long-acting (LA) injectable therapy with cabotegravir (CAB) and rilpivirine (RPV) is currently used as maintenance treatment for human immunodeficiency virus type 1, and has a low risk for virological failure (VF). Although the risk is low, the circumstances and impact of VF in the real-world setting merit further evaluation.Methods We performed an in-depth clinical, virological, and pharmacokinetic analysis on the reasons behind and the impact of VF during LA CAB/RPV therapy in 5 cases from the Netherlands. Genotypic resistance testing was performed after the occurrence of VF, and drug plasma (trough) concentrations were measured after VF was established and on any other samples to assess on-treatment drug levels. CAB and RPV drug levels that were below the first quartile of the population cutoff (<= Q1) were considered to be low.Results Five cases who were eligible for LA CAB/RPV experienced VF despite a low predicted risk at baseline. Genotypic resistance testing revealed extensive selection of nonnucleoside reverse transcriptase inhibitor-associated mutations in all cases, and integrase strand transfer inhibitor mutations in 4 cases. All cases displayed low drug levels of either CAB, RPV, or both during the treatment course, likely contributing to the occurrence of VF. In 3 cases, we were able to identify the potential mechanisms behind these low drug levels.Conclusions This is the first in-depth multiple case analysis of VF on LA CAB/RPV therapy in a real-world setting. Our observations stress the need to be aware for (evolving) risk factors and the yield of a comprehensive clinical, virological, and pharmacokinetic approach in case of failure. Long-acting injectable therapy with cabotegravir and rilpivirine is used as maintenance treatment for HIV-1 and has a low risk for virological failure. We present the first in-depth analysis of 5 cases of real-world virological failure on cabotegravir/rilpivirine therapy.