CROI 2016 Abstract eBook

Abstract Listing

Poster Abstracts

Methods: Two participants with HIV-1 viremia (plasma HIV-1 RNA >1,000 copies/ml) and 7 consecutive participants on suppressive ART were enrolled from the Pitt Lung HIV cohort and underwent bronchoscopy. Bronchoalveolar lavage (BAL) fluid was separated into cellular and cell-free fractions by centrifugation. AM were purified from BAL cells by plastic adherence with extensive washing. Plasma and peripheral blood mononuclear cells (PBMC) were also collected. HIV-1 RNA in the plasma and BAL supernatant were tested by qPCR targeting a highly-conserved region in HIV-1 pol . Cell-associated (CA) HIV-1 DNA and RNA in AM, total BAL cells, and PBMCs were assayed by qPCR targeting the same region of pol . To detect contaminating or ingested CD4+ T cells, T-cell receptor (TCR) RNA was tested by PCR. Results: Most subjects (89%) were male. The median age was 50 yrs and CD4 count was 735. In viremic individuals, CA HIV-1 DNA and RNA were detected in AM (median 1.5 and 2.5 log 10 copies/10 6 cells, respectively) and in PBMC (median 2.4 and 1.5, respectively). HIV-1 RNA in BAL supernatant was low compared to plasma (median 0.9 and 4.4 log 10 copies/ ml). Among subjects on ART, the median duration of viremia suppression was 3.6 yrs (range: 2.4, 12.3; n=7). CA HIV-1 DNA was detected in AM from 6 of 7 participants (median 1.7 log 10 copies/10 6 cells) and CA HIV-1 RNA in AM from 4 of 6 with detectable HIV-1 DNA (median 1.3). AM extracts were negative for TCR in 6 of 7 participants with samples available. By comparison, CA HIV-1 DNA was detected in PBMC from 6 of 6 individuals tested (median 2.5 log 10 copies/10 6 cells) and CA HIV-1 RNA in 2 of 6 tested (median 1.9). HIV-1 RNA in BAL supernatant was not detectable (< 0.3 copies/ml) despite low-level plasma HIV-1 RNA in 6 of 7 virally-suppressed participants (median 7.7 copies/ml). Conclusions: These results establish the persistence of HIV-infected AM in individuals on suppressive ART, which cannot be explained by contaminating or ingested CD4+T-cells. Studies are in progress to more fully characterize the proviruses in alveolar macrophages and their inducibility. 331 Evidence of HIV Evolution in Lymphatic and Cancer Tissues in ART+ Patients Rebecca Rose 1 ; Susanna L. Lamers 1 ; David J. Nolan 2 ; Debra L. Garcia 3 ; Marco Salemi 2 ; Ekaterina Maidji 4 ; Cheryl Stoddart 4 ; Elyse Singer 5 ; Michael S. McGrath 4 1 Bioinfoexperts, LLC, Thibodaux, LA, USA; 2 Emerging Pathogens Inst, Univ of Florida, Gainesville, FL, USA; 3 AIDS and Cancer Specimen Resource, San Francisco, CA, USA; 4 Univ of California San Francisco, San Francisco, CA, USA; 5 Natl Neurological AIDS Bank, Los Angeles, CA, USA Background: Anti-retroviral therapy (ART) is effective in reducing plasma HIV load to undetectable levels and restoring partial immunity. However, viral populations rapidly rebound once therapy is removed. Furthermore, HIV-associated co-morbidities, particularly certain cancers, remain a significant co-morbidity even with effective ART. We

hypothesized that HIV-infected macrophages in tumors may both protect HIV from ART and contribute to metastasis. Methods: The AIDS and Cancer Specimen Resource (ACSR) provided 26 post mortem tissues from three HIV+/ART+ patients with no detectable viral load. Two died frommetastatic lymphoma (Pt02, Pt04 ) , and one from lung cancer ( Pt05 ). Tissues were assessed for HIV using digital drop PCR. Single genome amplification was used to generate env and nef sequences. We inferred maximum-likelihood phylogenies and performed multiple statistical tests to investigate viral evolutionary patterns and compartmentalization. We used an HIV signal detection technique (RNAscope) combined with histological staining to visualize the cellular location of HIV RNA in tissues from Pt02 . Results: ddPCR was positive for all tissues. HIV env-nef DNA sequences were generated from 8/11, 3/6 and 9/9 tissues from each of the three patients, respectively. HIV RNA was also isolated from three tissues in Pt02 (lymph node tumor, cerebellum and aorta) and two tissues from Pt05 (lymph node and spleen). Maximum-likelihood phylogenies and statistical testing showed little evidence of viral compartmentalization among tissues for RNA and DNA. Considering that the patients had been on ART, some clades showed surprisingly high diversity and evidence of on-going evolution, while others were consistent with a pattern of clonal expansion. RNAscope for Pt02 showed that RNA expression of HIV gag-pol was clustered and co-localized with CD163+/CD68+macrophages in tumor lymph node and the cerebellum, with lymphoma involvement due to meningeal metastasis.. Infiltrating macrophages surrounded HIV infected cells ( Figure ). Conclusions: Our results suggest that lymphatic and cancer tissues may offer a privileged environment for persistent HIV replication within macrophages during ART and may promote tumor growth and metastasis. Two distinct patterns of tissue virus evolution suggest that different modes of replication/spread underlie persistence: ART-resistant HIV infected macrophages with persistent evolution/spread and clonal expansion of HIV infected cell populations.

Poster Abstracts

332 Role of Transitional-Memory T Cells in Productive HIV Reservoir in ULTRASTOP Patients Chiraz Hamimi 1 ; Sidonie Lambert 2 ; Ruxandra Calin 3 ; Guislaine Carcelain 1 ;Yasmine Dudoit 3 ; Lambert Assoumou 2 ;Vincent Calvez 2 ; Dominique Costagliola 2 ; Christine Katlama 2 ; Brigitte Autran 1 1 INSERM, UMR-S 1135, Paris, France; 2 Sorbonne Univs, Paris, France; 3 APHP, Pitié-Salpêtrière Univ Hosp, Paris, France Background: The Ultrastop clinical study aims at evaluating frequency and parameters of HIV-remission after treatment interruption (TI) in early-chronic treated patients with an ultra-low HIV reservoir. The success of drug free remission was 10%. Half patients carried protective HLA-B*27 or B*57 and CD8-T-cell responses were weak at baseline in all patients including the post treatment controller (PTC). Here, we analysed the reservoir dynamic and HIV-reactivation upon TI in the ULTRASTOP Cohort. Methods: Prospective study of treatment-interruption (TI) followed by treatment-resumption (RXR) in case of immuno-virological failure (pVL>400 cp/mL or CD4<400/mm3) in ten early-chronic treated patients enrolled with ultralow HIV-DNA (<66 cp/106 PBMC). Monitoring of immune-virological parameters was performed from D0 until W48 off-ART and at W4/W12/W24 after RxR. Dynamics of HIV reservoirs were quantified in PBMCs and sorted resting naïve, central-, transitional- and effector-memory CD4 T cell subsets at D0, RxR and W24 post RxR using ultrasensitive RT-PCR. Reactivation of HIV-1 from CD4 T-cell was performed by anti CD3/CD28, IL2 and IL7 stimulation Results: One HLA-B*27 + CCR5 w/w patient controlled viremia up to W56 off-ART and was defined as PTC. 9/10 patients, 4 being HLA-B*27 or B*57+, had prompt plasma viral rebound (W2-12) accompanied by an increase in cell-associated HIV-DNA which returned to baseline levels at W24 post RxR. At baseline, HIV-DNA distribution did not differ between resting naïve and memory subsets in all patients, and was below the threshold in the PTC’s TTM and TEM compartments. HIV-1 was reactivated in-vitro upon total CD4 T-cells stimulation in only 3 patients that displayed the highest HIV-DNA levels at baseline, but not in the PTC. At RxR increase in HIV-DNA levels involved the TCM and TTM subsets with a preferential contribution of TTM to the CD4 compartment. HIV became inducible in-vitro in total CD4 T-cells from all non-controllers as well as in PTC’s CD4 T cells. RxR levels of HIV-RNA reactivation correlated with both peak of pVL (p 0.006) and HIV-DNA levels in PBMC (p 0.03) and in TTM (p 0.01). Conclusions: In a highly selected population of early-chronic treated patients with ultra-low HIV reservoir, a success rate of 10% after TI was observed. The relatively short-lived transitional-memory T cells compartment appears to play a key role in contribution to virus production both in vitro and in vivo. these findings open new perspectives in targeting the productive HIV reservoir 333 Drug Activity in PrEP Breakthroughs Has a Transient Effect on SHIV DNA Reservoirs Mian-er Cong; Chou-Pong Pau;Walid Heneine; Gerardo Garcia-Lerma CDC, Atlanta, GA, USA Background: Infection of macaques with simian HIV (SHIV) during concurrent pre-exposure prophylaxis (PrEP) with emtricitabine (FTC) and tenofovir disoproxil fumarate (TDF) is associated with reduced acute plasma viremias and limited virus diversity, thus providing a unique model to assess long-term effects of acute antiretroviral treatment. Here, we investigated the effect of PrEP on acute SHIV DNA dynamics in peripheral blood mononuclear cells (PBMCs), and on the size of the persistent virus reservoir in lymphoid tissues.

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