CROI 2025 Abstract eBook

Abstract eBook

Poster Abstracts

372

Dynamic Changes in the Peripheral Blood Monocytes Following Long-Acting Injectable CAB/RPV in PLWH Maria Antonella Zingaropoli 1 , Eeva Tortellini 1 , Maria Silvia Guardiani 1 , Anna Carraro 1 , Federica F. D. Dominelli 1 , Cosmo Del Borgo 2 , Raffaella Marocco 2 , Giulia Mancarella 1 , Lorenzo Ansaldo 2 , Maria Rosa Ciardi 1 , Claudio Maria Mastroianni 1 , Miriam Lichtner 1 1 University Sapienza, Rome, Italy, 2 Santa Maria Goretti Hospital, Latina, Italy Background: The advent of long-acting (LA) injectable ART has transformed the conceptualization of ART. However, there is a need for Real World Data on the impact of this novel approach on peripheral inflammation. The aim of this study was to investigate the dynamic changes in monocyte/macrophage subsets and their soluble activation markers in PLWH switching to LA injectable ART (CAB/RPV). Methods: The present study investigated the percentages of peripheral blood monocyte/macrophage and dendritic cell subsets (HLA-DR, CD14, CD16, CD11c, CD123) in samples obtained from PLWH before CAB/RPV (T0) and after six (T6) and twelve months (T12) by flow cytometry. Moreover, a longitudinal evaluation of monocyte/macrophage activation markers (sCD14 and sCD163) was conducted. Subsequently, each time point was compared to a control group (healthy donors, HDs). Results: A total of 30 aviremic PLWH switching to CAB/RPV, and 32 HDs were enrolled. All the enrolled PLWH were ART-experienced (12 dual/18 no dual) and 93.3% had a viral load of <50 copies/mL prior to initiating LA ART. At T12, no virological failure was observed. In comparison to HDs, PLWH exhibited higher percentages of intermediate monocytes (p=0.0037), non-classical monocytes (p<0.0001), M-DC8 (p=0.0007) and elevated plasma levels of sCD14 (p<0.0001) and sCD163 (p<0.0001) at each time point. The stratification of PLWH according to previous ART revealed no differences in the percentages of monocyte/ macrophage subsets or in the plasma levels of sCD14 and sCD163 between dual and no dual therapy. Overall, at both T6 and T12, a significant reduction in the percentage of non classical monocytes compared to T0 was observed (p=0.0054 and p=0.0017, respectively). No significant differences were observed in the percentages of classical monocytes, intermediate monocytes, M-DC8, pDCs and mDCs. Furthermore, a reduction in sCD14 plasma levels was observed at both T6 and T12 in comparison to T0 (p=0.0006 and p=0.0237, respectively). No differences in sCD163 plasma levels were observed (see Table). Conclusions: The significant reduction in inflammation and peripheral immune activation of the monocyte compartment observed after 12 months of LA injectable CAB/RPV underlines that the potential contribution of these cells to the viral reservoir and persistence may vary with therapy. The constant drug concentrations provided by CAB/RPV injections appear to be sufficient for sustained virological suppression and may affect the monocyte/macrophage compartment in PLWH.

Methods: We measured ccf-mtDNA levels in the AIDS Linked to the Intravenous Experience (ALIVE) (N=890) and Multicenter AIDS Cohort Study (MACS)(N=427). Using ultrasensitive digital PCR, we quantified short and long ccf-mtDNA fragments in serum samples. Measurements were taken at intervals of 1, 3, or 5 years. Levels of ccf-mtDNA were compared by HIV serostatus and CD4 T-cell counts (<350 vs. ≥350 cells/mm³), using mixed-effect linear regression models that controlled for demographics, BMI, active injection drug use, and alcohol use. We also measured ccf-mtDNA levels in non-human primates (n=6) before and after SIV infection and following antiretroviral therapy (ART) initiation. Samples were drawn at baseline, peak viremia (2 weeks post-infection), pre-ART (5 weeks post-infection), and 3- and 6-months post-ART initiation. Results: In both cohorts, PWH had significantly lower levels of both short and long ccf-mtDNA fragments compared to HIV-negative individuals. The difference was more pronounced in the ALIVE cohort (0.119 log scale units lower) than in the MACS cohort (0.094 units lower) (both p<0.05). Furthermore, individuals with lower CD4 T-cell counts (<350 cells/mm³) exhibited further reductions in ccf-mtDNA levels, with significant reductions in both short and long fragments in the ALIVE cohort and short fragments in the MACS cohort (Figure Panel A). In nonhuman primates, short ccf-mtDNA levels increased shortly after infection, peaking just before ART initiation (p=0.001), and subsequently declined, reaching levels below baseline after 6 months on ART. Long fragments remained stable during the early post-ART phase but declined significantly by 6 months (p=0.02, Figure Panel B). Conclusions: Contrary to the initial hypothesis, ART-suppressed HIV infection is associated with lower levels of both short and long ccf-mtDNA fragments, following an initial increase during acute infection. These findings underscore the need for further investigation into the mechanisms regulating ccf-mtDNA in people living with HIV and its potential role in aging-related diseases. The Molecular Cargo of Exosomes Is Able to Discriminate Between PLWH With or Without NADCs Jose M. Benito 1 , Asier Ureta 1 , Clara Restrepo 1 , Manuel Pedregal 2 , Héctor Callata 2 , Sara Nistal 3 , Alejandro Velastegui 3 , Edinson Caviedes 4 , Hector Peinado 5 , Manuel J. Gómez 6 , Fátima Sánchez-Cabo 6 , Alberto Alcázar 7 , Emma Martínez-Alonso 7 , Jesús García-Foncillas 8 , Norma Rallón 1 1 Instituto de Investigación Sanitaria Fundación Jiménez Díaz, Madrid, Spain, 2 Hospital Universitario Fundación Jiménez Díaz, Madrid, Spain, 3 Hospital Universitario Rey Juan Carlos, Madrid, Spain, 4 Hospital Universitario Infanta Elena,Valdemoro, Spain, 5 Spanish National Cancer Research Centre (CNIO), Madrid, Spain, 6 Centro Nacional de Investigaciones Cardiovasculares, Madrid, Spain, 7 Hospital Universitario Ramon y Cajal, Madrid, Spain, 8 Universidad Autónoma de Madrid, Madrid, Spain Background: Non-Aids Defining Cancers (NADCs) are the main cause of morbidity and death from non-AIDS events (ENOS) in PLWH, whose death rate is higher than in the general population without HIV infection but who suffer from these same NADCs. However, the mechanisms that explain this are unknown. It has been suggested that exosomes derived from HIV infected cells can promote and enhance cancer in vitro . Herein, we have analyzed the proteomic and miRNA cargo of exosomes from PLWH with active NADC to identify a molecular profile associated to this condition. Methods: Forty-two participants were included: 11 PLWH on cART (HIV + NADCs - ), 11 on cART with NADC (HIV + NADCs + ), 10 HIV-uninfected with NADCs (HIV - NADCs + ), and 10 healthy volunteers (HC). Size Exclusion Chromatography was employed to isolate plasma exosomes. Protein and miRNAs content of exosomes isolated from plasma were analyzed by liquid chromatography coupled with mass spectrometry (proteins), and by RNAseq in a Miseq System (miRNAs). Canonical discriminant analysis (CDA) was used to identify the molecular profile (miRNA and/or proteins) in the exosome cargo from PLWH with NADCs compared to the other study groups. Results: A total 599 miRNAs and 317 proteins were detected in the exosomes cargo of study groups. Only those proteins or miRNAs showing differential expression (DE) between the different study groups (130 miRNAS and 133 proteins) were included in the CDA. The CDA was able to completely (100%) discriminate between the four study groups. The CDA for pairs of groups (six different models) was also able to perfectly (100%) discriminate between participants in each group. Moreover, the profile of miRNAs and proteins included in each of the six CDA models was exclusive of each model, suggesting a specific profile of exosomes cargo associated to each study group. Interestingly, the CDA models discriminating between HIV + NADCs + and HIV - The figure, table, or graphic for this abstract has been removed.

Poster Abstracts

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Changes in Serum Level of Cell-Free Mitochondrial DNA Fragments in Acute and Chronic HIV Infection Jing Sun 1 , Lolita S. Nidadavolu 2 , Hsing-Yu Hsu 1 , Jacquie Astemborski 1 , Jonah B. Sacha 3 , Paul Kievit 3 , Charles Roberts 3 , Gregory Kirk 4 , Todd Brown 2 , Peter Abadir 2 , for the ALIVE and MWCCS Cohorts 1 The Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA, 2 The Johns Hopkins University School of Medicine, Baltimore, MD, USA, 3 Oregon Health and Science University, Portland, OR, USA, 4 The Johns Hopkins University, Baltimore, MD, USA Background: Circulating cell-free mitochondrial DNA (ccf-mtDNA) fragments are released into the bloodstream following cell death, can activate the innate immune system, and have been associated with aging-related outcomes, including frailty, in the general population. Data on ccf-mtDNA levels during both acute and chronic phases of HIV infection remain limited. We hypothesized that HIV infection is associated with higher ccf-mtDNA.

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CROI 2025