CROI 2015 Program and Abstracts

Abstract Listing

Poster Abstracts

Liver Macrophages Harbor Infectious HIV-1 for Prolonged Durations and Despite cART.

Whole liver tissue was obtained from HIV-1 infected individuals at time of death (*) and during liver transplantation (^). After whole cell lysis, samples were tested for HIV-1 proviral DNA. Cell numbers were estimated using ERV3 DNA quantification from the same samples. The table indicates the latest available CD4+ T cell count, cART status, and plasma HIV-1 RNA before the recovery of liver tissues. a - Clinical laboratory values that were contemporaneous for LT. § - Proviral DNA was quantified using a qPCR assay that was adapted to detect tissue HIV. *liver samples were obtained from NDRI; HIV-1 RNA, cART status, HCV and HBV status are reported by NDRI. Data that was unavailable is indicated by“-”. HIV-1 RNA, cART status, HCV and HBV status were available through clinical testing for LT01 and LT02. TDF – tenofovir disoproxil fumarate; ddI – didanosine; NFV – nelfinavir; FTC – emtricitabine; EFV – efavirenz; Ral – raltegravir; ABC – abacavir; 3TC – lamivudine; DTG – dolutegravir; UD – undetectable using a standard clinical assay. ND- not done.LOD- limit of detection Conclusions: These data provide strong evidence that LM, the largest TRM population, release infectious HIV-1 after a prolonged duration; therefore, LMmay represent an important reservoir of HIV infection and potential impediment to cure. 381 Large-Scale Analysis of HIV-1 Integration Sites in Untreated and Treated Patients Stephanie Laufs 1 ; Diana Schenkwein 1 ; Neeltje Kootstra 2 ; Frank A. Giordano 5 ; Christoph Stephan 3 ; Hans-Georg Kraeusslich 4 ;Winfried Kern 6 ; Susanne Usadel 6 ; Manfred Schmidt 1 ; Christof von Kalle 1 1 National Center for Tumor Diseases; German Cancer Research Center, Heidelberg, Germany; 2 Sanquin Research, Landsteiner Laboratory, and Center for Infectious Diseases and Immunity Amsterdam, Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands; 3 University Hospital Frankfurt, Frankfurt, Germany; 4 Department of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany; 5 Heidelberg University, Heidelberg, Germany; 6 Division of Infectious Diseases, Department of Medicine, Albert-Ludwigs-University Center for Infectious Diseases & Travel Medicine, and IFB-Center for Chronic Immunodeficiency, University Hospital, Freiburg, Germany Background: The HIV/AIDS pandemic remains an important threat to global health. Although cART (combination antiretroviral therapy) induces a rapid decline in plasma HIV-1 RNA, persistence of latent forms of HIV-1 displays the major hurdle on the way to eradicating the infection. Recent data indicate that at least a subset of patients harbor a sufficiently suppressed viral reservoir which is not able to fuel viral rebound upon therapy discontinuation. These so-called functionally cured patients provide new hope for an HIV cure by the elimination of replication-competent HIV-1 reservoirs. Based on our experience from the analysis of more than 1 million lentivirus vector insertion sites (IS) we hypothesized that significant portions of inserted provirus in patients might be non-randomly distributed and/or clonally stable and/or within illegitimate, non-canonical LTR boundaries. Methods: To study wtHIV IS in human patients, we determined genomic/proviral flanking regions in untreated (n=39) and cART treated (n=32, including amongst others an integrase inhibitor) patient cohorts using linear amplification-mediated PCR (LAM-PCR) in a longitudinal analysis. The clonal inventory of infected cells was obtained using next-generation sequencing combined with a novel double barcoding system in combination with innovative bioinformatic analysis. Integrome analysis was done to quantify chromosomal distribution of IS, illegitimate non-integrase mediated HIV-1 integration and clonal persistence indicative of virus-host genome interactions. Results: Our data describe for the first time the existence of LTR micro-deletions in wtHIV infected CD4 positive patient cells. To generate the IS profile we identified more than 2500 integration sites fromwtHIV-1 infected patient samples pre and post therapy and identified common IS. To monitor longitudinal IS distribution, we performed high-throughput IS analysis at up to eight different time points up to 330 days after therapy. Identical IS were found at multiple time points proving the existence of a persisting reservoir of dividing infected cells in patients. Conclusions: This finding is in line with recent reports where the authors postulate that the location of HIV ISs can promote the expansion and persistence on HIV infected cells (Maldarelli et al. 2014, Wagner et al. 2014). In addition our data proof LTR micro-deletions in wtHIV infected patient cells and focus on HIV integration in humans on a large cohort of treated and untreated patient samples.

Poster Abstracts

282

CROI 2015