CROI 2019 Abstract eBook

Abstract eBook

Poster Abstracts

Conclusion: g-NK cells are more frequent in HIV-infected patients compared to controls and may contribute to low CD4 counts in HIV patients and poor recovery during ART. g-NK cells may be a useful biomarker for predicting how the CD4+ T cell population may recover during HIV treatment.

1 University of California San Diego, San Diego, CA, USA Background: HIV persistence in cellular reservoirs is the main barrier to a cure. The size and composition of HIV DNA populations in solid tissues during suppressive antiretroviral therapy (ART) is not well characterized. Methods: We examined the distribution and genotypic composition of the HIV DNA populations across paired post-mortem tissues from one virally suppressed person living with HIV (PLWH) from the Last Gift (LG) Cohort. The LG cohort enrolls altruistic, terminally-ill PLWH, who are closely followed until the time of death and donate their “whole-body” for HIV research. Blood and tissues are collected by rapid-autopsy and cryopreserved within 6h from death. From each sample, we extracted total DNA and quantified HIV DNA (gag) levels by droplet- digital PCR. The genotypic composition of the HIV DNA in tissues was evaluated using a high-throughput single genome amplification and the PacBio platform to deeply sequence full-length HIV envelope (FL HIV-env). Results: The participant was a 72-year-old man with chronic HIV infection and metastatic pancreatic cancer. He enrolled in the LG study 5 months prior death. He was on ART and had undetectable HIV RNA in blood plasma, up to 7 hours before death (<20 copies/ml). From 26 paired post-mortem tissues, HIV DNA was detected in 24 samples, including brain (3-11 cps HIV/106 cells), gastrointestinal (45-211), urogenital tract (46-377), lymphoid (22-243) and adipose (13-874) tissues. HIV DNA was undetectable in parietal and motor cortex. We obtained 107 individual FL HIV-env sequences across 10 tissues (median 10.7 sequences/tissue), of which 60 were unique. The maximum likelihood phylogeny (figure) showed a deep divergence, segregating the tree into two lineages, which differed by co-receptor tropism, based on in silico tropism prediction (geno2pheno). Interestingly, 100 FL HIV-env sequences were genetically intact, while 7 sequences were non-functional, with major deletions, frameshifts, and stop codon mutations. HIV-env migration appeared to be extensive, with many identical sequences sampled in multiple body tissues. Conclusion: HIV DNA was detected in most body tissues despite long-term ART and confirmed undetectable HIV RNA at the time of death. Based on the FL HIV-env sequencing, most HIV reservoirs appeared to be intact provirus and may present different viral tropisms. The LG cohort poses a unique opportunity to characterize the HIV reservoirs in anatomic compartments, which is crucial to provide insights for future HIV cure strategies.

Poster Abstracts

326 LIPIDOMIC FINGERPRINTING TO IDENTIFY HIV RESERVOIRS IN VITRO AND IN VIVO Christina Gavegnano 1 , Yong Jiang 1 , Sijia Tao 1 , Ruby Kleinbard 1 , Anush Arakelyan 2 , Andrea Lisco 3 , Irini Sereti 3 , Leonid Margolis 2 , Raymond F. Schinazi 1 1 Emory University, Atlanta, GA, USA, 2 National Institute of Child Health and Human Development, Bethesda, MD, USA, 3 National Institute of Allergy and Infectious Diseases, Bethesda, MD, USA Background: One of the most important questions in designing cure-based strategies in HIV research is the origin(s) of the reservoir. It is well established that HIV rebounds upon withdrawal of Highly Active Antiretroviral Therapy (HAART) leading to viral breakthrough and HIV envelopes contain lipids originating from the membrane of the virus producing cells. Thus, an innovative highly sensitive, clinical sample-validated Virus Lipidomic Mapping (VLM) assay was developed to accurately quantify lipid compositions of HIV-1 virions (< 5 copies/mL) and host cells, allowing virus to be mapped to host cell origin. Methods: Primary human CD4+ T cells or monocytes were isolated (healthy buffy coats; magnetic sorting; n = 12). HIV-1BaL was used to infect CD4+ T cells or differentiated macrophages (MΦ). HIV from CD4+ T cell or MΦ origin (n = 4) was captured from viremic plasma of HIV+ individuals with magnetic nanoparticles (MNP) coupled to Ab against T cell or MΦ-specific proteins incorporated into virions followed by virus purification and exosome removal. Lipids were extracted from cells; profiles, abundance and ratios (cells and virus) were determined by LC-MS/MS (Q-Exactive Plus). Results: For all donors, the cellular lipid profiles for CD4+ T cells were distinct fromMΦ lipid profiles. Virions grown in each cell type contained unique lipids that match unique lipids on host cell origin. Distinctive profiles for MΦ cellular and virion lipids included monoetherphosphatidylcholine (MePc), ceramides (Cer), glucosylceramide (CerG1), diacylglycerol (DG), phosphatidylinositol (PI), phosphatidylcholine (PC, phosphatidylethanolamine (PE), sphingomyelins (SM), and triglyceride (TG); unique profiles for T-cell virion and cellular lipids include subspecies SM(d35) and CerG1(d18). For all donor samples, virions that carried specific T-cell proteins showed unique lipid profiles compared to virions that carried MΦ-specific proteins; Cer (d17, G1d18), DG(53.1), MePC(31), PC(34), PE(42), PI(18), SM(d35), and ChE(18). Conclusion: Conclusions: For the first time, the VLMmethod 1) identified host cell origin of persistent HIV-1 in vivo, even with low-prevalence populations of mixed virions, and 2) determined unique lipid profiles for virions from T-cells versus MΦ, which match cellular lipid profiles on host cell origin. This information provides a foundation for cure-based strategies to identify and eliminate key cells harboring persistent HIV not eliminated by HAART. 327 CHARACTERIZING THE HIV DNA RESERVOIRS IN WHOLE-BODY TISSUES IN THE “LAST GIFT” COHORT Michelli Faria de Oliveira 1 , Benjamin Murrell 1 , Thomas Vollbrecht 1 , Susanna Concha-Garcia 1 , Venkatesh Kumar 1 , Magali Porrachia 1 , Brianna Scott 1 , Laura Layman 1 , Caroline Ignacio 1 , Sara Gianella 1 , Davey M. Smith 1

328 CD4 T-CELL SUBSETS RESPOND DIFFERENTLY TO LATENCY REVERSAL AGENTS Judith Grau-Expósito 1 , Laura Luque-Ballesteros 1 , Adrià Curran 2 , Joaquin Burgos 2 , Esteban Ribera 2 , Ariadna Torrella 2 , Bibiana Planas 2 , Rosa Badía 2 , Vicenç Falcó 2 , María J. Buzón 1

CROI 2019 119