CROI 2025 Abstract eBook

Abstract eBook

Poster Abstracts

387

Ultra-Low Level Soluble p24 Is Associated With Inflammation in People With HIV on Suppressive ART Matteo Augello, Alessandra Mingione, Valeria Bono, Roberta Rovito, Camilla Tincati, Filippo Martinelli Boneschi, Giulia Marchetti University of Milan, Milan, Italy Background: Despite suppressive ART, residual inflammation persists in people with HIV (PWH), thus driving higher risk of SNAEs and mortality. Circulating HIV proteins have been detected in some suppressed PWH, as an expression of the persistence of translation-competent proviruses. We therefore seek to investigate whether peripheral inflammation and mortality risk are associated with production of p24 in PWH on suppressive ART. Methods: PWH on suppressive ART (HIV-RNA <20 cp/mL) and age/sex matched healthy controls without HIV were enrolled. We cross-sectionally measured ultra-low level serum p24 (Simoa), 12 plasma markers of inflammaging (IP-10/TNF-α/IL-2/IL-4/IL-17A/IL-6/IL-8/sCD14/sCD163/TIMP-1/ GDF-15/ROMO-1) [Luminex], and activated (CD38+) CD8+ T-cells (flow cytometry). Mortality risk was estimated by VACS Index 2.0. Mann-Whitney test and linear regression were used for analysis. Results: 120 PWH and 20 controls were recruited. PWH (83.3% males) had a median age of 54 (IQR 47–59) years. They were on ART for 10 (6–19) years, with a current CD4 T-cell count of 725 (504–908) cells/ μ L. Compared to controls, PWH showed higher IP-10/TNF-α/IL-17A/IL-8/sCD14/ sCD163/TIMP-1/GDF-15. VACS Index 2.0 was positively associated with IP-10/TNF-α/sCD14/sCD163/TIMP-1/GDF-15/CD8+CD38+. Yet, by fitting a multivariable regression model including all these seven markers, only IP-10, GDF-15 and CD8+CD38+ confirmed independently associated with VACS Index 2.0. By setting the p24 positivity cut-off at 0.025 pg/mL based on the mean+3SD of controls, 25 (20.8%) PWH had detectable p24 (median 0.0516 pg/mL; range 0.0264–0.8280 pg/mL) ( Fig.1A ). When assessing inflammaging markers in PWH according to p24 detectability, p24+ PWH showed significantly higher IP-10 levels. Accordingly, soluble p24 was positively associated with IP-10 at both univariable and multivariable (adjusted for sex, age, CD4 T-cell count, ART duration) regression analysis ( Fig.1B ). Conversely, no association was found between soluble p24 and CD38+CD8+ T-cells. Conclusions: Despite long-term suppressive ART, ultra-low level soluble p24 is detectable in ~20% PWH and associated with higher circulating IP-10, a pro inflammatory chemokine involved in the recruitment of innate and adaptive immune cells into sites of inflammation. Given the positive association between IP-10 and VACS Index 2.0, a measure of physiologic frailty and risk of death, these data suggest that p24-driven inflammation may contribute to SNAEs and mortality in virologically-suppressed PWH. The figure, table, or graphic for this abstract has been removed. Inflammatory and Microbial Signatures Linked to Cardiovascular Outcomes in People With/Without HIV Rachel MacCann 1 , Gurvin Saini 1 , Jesvin Xavier 1 , Alejandro Abner Garcia Leon 1 , Manon Vanbellinghen 2 , Neeltje Koostra 2 , Junhui Li 3 , Padraig McGettrick 1 , Alan Landay 4 , Aoife Cotter 1 , Alan Winston 5 , Peter Reiss 2 , Paul W. O'Toole 1 , Sabin Caroline 6 , Mallon W. G. Paddy 1 1 University College Dublin, Dublin, Ireland, 2 Amsterdam University Medical Centers, Amsterdam, Netherlands, 3 University College Cork, Cork, Ireland, 4 University of Texas Medical Branch, Galveston, TX, USA, 5 Imperial College London, London, UK, 6 University College London, London, UK Background: Chronic inflammation in people with HIV (PWH) is linked to cardiovascular disease (CVD). We previously described distinct inflammatory clusters from 24 biomarkers that were associated with CVD. We aimed to validate these results in an independent cohort and assess whether additional biomarkers or gut microbiome signatures could enhance CVD prediction in PWH. Methods: In plasma from people with and without HIV from three international cohorts we measured 55 biomarkers by quantitative immunoassays. Principal component analysis (PCA) and hierarchical clustering (HC) was performed on the original 24 biomarkers and a recursive feature addition (RFA) model used to assess the stepwise addition of 31 additional biomarkers, selecting those that enhanced the model for repeat PCA and HC. Stool microbiome was analysed by 16S rRNA sequencing, with differentially abundant species identified by Analysis of Compositions of Microbiomes with Bias Correction (ANCOM-BC) and correlated to biomarkers using Spearman correlation. Multinomial logistic

regression explored associations between clusters and CVD outcomes. Data are median (IQR) unless stated. Results: We included 408 participants (age 50 (43–58) years; 83% male; 68% White; 78% PWH). Consistent with previous studies, initial PCA of 24 biomarkers again revealed 3 clusters, with cluster 3 (n= 41) showing elevated markers of inflammation (IL1-β, IL-6, TNF-α), immune regulation (TSLP, MIP1-α), and T-cell differentiation (IL2, IL12, IFN-γ). Cluster 3 participants were older (51 (47-55) years), had higher BMI (27.5 (24.4–29.2)kg/m2), higher triglycerides (1.98 (1.23- 2.62) mmol/L), and higher CVD prevalence (n= 6 (39%)). After RFA, 13 of the 31 additional biomarkers enhanced the PCA, with cluster 3 now including GM-CSF, IFN-α2a, TGF-α, and Thrombopoietin, with a stronger CVD association (OR: 2.13 (95% CI: 1.03, 4.4), p= 0.04). In cluster 3, ANCOM-BC revealed 5 enriched and 7 depleted microbial species . Within cluster 3, IFN-l2, MIP-1 α, MCP-1, CD40-ligand and TGF-α all correlated with depletion of short-chain fatty acid producing species ( B. coprocola , B. massiliensis , F. mortiferum , D. invisus , Figure ) that are linked to regulation of inflammation and CVD. Conclusions: This analysis again confirmed distinct inflammatory clusters associated with CVD in an independent cohort of PWH. Clusters enhanced with additional biomarkers better predicted CVD and identified lined gut microbiome signatures linked to CVD. Methamphetamine Use Disorder in ART PWH Is Associated With Elevated NLRP3 Inflammasome Activation Felipe Ten-Caten 1 , Fernanda C. Coirada 1 , Alexander L. Silva-Junior 1 , Caroline H. Sheikhzadeh 2 , Sannidhi Sarvadhavabhatla 2 , Alton Barbehenn 2 , Poonam Vohra 2 , Cassandra Yun 2 , Nitasha A. Kumar 2 , Paula J. Lum 2 , Kara L. Lynch 2 , Steven G. Deeks 2 , Rafick P. Sekaly 1 , Sulggi Lee 2 , Susan P. Ribeiro 1 1 Emory University, Atlanta, GA, USA, 2 University of California San Francisco, San Francisco, CA, USA Background: Despite effective antiretroviral therapy (ART), people with HIV (PWH) experience higher morbidity and mortality rates than people without HIV due to ongoing inflammation from persistent HIV. Methamphetamine (MA) use disorder among PWH is linked to lower ART adherence, increased viral resistance and poorer clinical outcomes. We recently demonstrated that MA exposure during ART is associated with increased HIV transcription. Here, we performed single cell analyses to determine signaling pathways by which MA induces immune dysfunction, potentially driving viral transcription in PWH even during ART suppression. Methods: Longitudinal inguinal lymph node (LN) samples were obtained from 10 ART-suppressed PWH with MA use disorder. Plasma MA concentrations were analyzed in relation to single cell gene (scRNA-seq) in LNs and protein (flow cytometry) expression from PBMCs. Linear regression models were used for comparisons. Results: MA was associated with significantly higher expression of NLRP3 and IL-1β signaling genes from monocytes, dendritic cells (DCs), and neutrophils. Participants with high plasma concentrations of MA had significant upregulation of “TNF-α signaling via NF-κB” and “Inflammatory response” pathways in DCs, CD8 T cells, NKs, and granulocytes. B and plasma cells demonstrated significant upregulation of the “IFN alpha and gamma responses”. Higher plasma concentrations of MA was also associated to decreased expression of chromatin modifier genes in DCs, monocytes, and plasma cells, consistent with open chromatin state in these cell types. Flow cytometry data from CD4+ T cells demonstrated enhanced chromatin accessibility in participants with high MA concentrations. Conclusions: Leveraging a unique cohort of PWH on ART with and without MA use disorder, we demonstrate that MA is associated with increased NLRP3 inflammasome activation, potentially driven by epigenetic changes to cell specific subsets. Additionally, MA was associated with enhanced chromatin accessibility in CD4+ T cells, which, along with increased inflammatory signatures in all immune cell subsets, supports our prior findings that MA induces increased HIV residual transcription. Finally, we demonstrate that MA may induce humoral dysfunction, with clinical implications for decreased antibody responses in this population. These data highlight potential novel mechanisms by which MA may promote immune dysfunction in PWH, with implications for future strategies at reducing co-morbidities in this population. The figure, table, or graphic for this abstract has been removed.

Poster Abstracts

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CROI 2025