CROI 2017 Abstract e-Book

Abstract eBook

Poster and Themed Discussion Abstracts

217 ALCOHOL CONSUMPTION INCREASES SIV FOUNDER VIRUS DIVERSITY TO ALTER DISEASE COURSE

Spencer Robichaux , Angela M. Amedee Louisiana State Univ, New Orleans, LA, USA

Background: Alcohol abuse is a highly prevalent comorbid condition in the HIV-infected population, which increases the risk of HIV transmission and adversely affects disease progression. Using Simian Immunodeficiency Virus (SIV) infected rhesus macaques exposed to chronic binge alcohol (CBA), our studies have shown that CBA alters multiple aspects of SIV infection and disease by increasing CD4+ target cell proportions in the GALT prior to SIV, increasing susceptibility to rectal mucosal SIV challenge, increasing set point viremia, and decreasing median time to death. Our current studies sought to provide insights into virus:host interactions that affect early replication dynamics and influence disease progression in CBA animals. Methods: We characterized the founder virus populations (FV) and replication kinetics in 14 CBA and 12 control rhesus macaques inoculated rectally with SIVmac251. FV were assessed using classic single genome amplification of SIV-gp160 and a novel next generation sequencing assay targeting V1-V2. Results: The numbers FV genotypes transmitted to individual animals did not differ between CBA and controls, with 1-3 genotypes observed (mean of 1.67, 1.86 respectively). However, genotypic analyses of FV populations showed that CBA animals expressed a more diverse founder virus population as compared to controls (p<0.001). At viral set point (10 weeks p.i.), CBA-treated animals showed significantly higher levels of viral RNA in both plasma and lymph node reservoirs (p<0.05), while similar lymph node proviral DNA burdens were observed CBA and control animals. Further analysis at set point of intra-animal evolutionary changes in viral genotypes showed that control animals exhibited a minor increase in diversity, while CBA animals showed a significant reduction in diversity (p<0.001). Conclusion: These observations suggest that CBA mitigates early host selective pressures resulting in the establishment of a more genetically diverse founder viral population. At set point, increased viral RNA levels in tissue reservoirs and higher plasma viremia in CBA animals, concomitant with a decrease in diversity of expressed viral genotypes. 218 GORILLA ENTERIC VIROME DYNAMICS IN ASSOCIATION WITH SIV INFECTION Background: The ancestors of the four groups of HIV-1 have been described in distinct communities of chimpanzee and gorilla from Cameroon. The infection of chimpanzees with SIVcpz has already been associated with progression to an AIDS-like disease. The expansion of the enteric virome associated with disease progression in an AIDS-like context has also been described in the rhesus macaques model, not observed in the non-pathogenic natural SIV infection of African green monkey. The aim of this study was to identify and compare enteric viromes of SIV-infected and uninfected gorillas, and to assess their relationship with SIVgor pathogenesis in their natural host. Methods: We analyzed 22 fecal samples from 11 SIV-infected and 11 uninfected gorillas from Cameroon. NGS was carried out in a HiSeq 2500 Illumina platform and analyses were conducted with an in-house pipeline using the programs FastQC, Sickle-Master, BlastX - Viral Database from GenBank/NCBI, MEGAN5 and RStudio. Results: The viral families Bromoviridae, Myoviridae, Podoviridae, Rhabdoviridae and Tymoviridae were statistically (p<0.01) more abundant in the uninfected group, whereas Alloherpesviridae, Herpesviridae, Reoviridae and Siphoviridae families were more abundant (p<0.01) in the SIV-infected group. Also, two distinct clusters were recognizable when a 1,000 cp/mL cutoff of SIVgor viral load in faeces (ranged in 655 to 31,497 cp/mL) was used to assess within-group diversity. The 6 samples with higher SIVgor viral load showed Mimiviridae, Myoviridae, Parvoviridae, Phycodnaviridae, Podoviridae, Reoviridae and Retroviridae statistically (p≤0.05) more abundant families and the 5 samples with smaller viral load showed Adenoviridae, Alloherpesviridae, Inoviridae, Siphoviridae and Unclassified Phages statistically (p<0.05) more abundant taxa. Finally, we are able to detect adenovirus-assigned reads (virus associated with intestinal disease in rhesus monkeys with advanced AIDS) only in the SIV-infected samples that belong to the smaller viral load group with known infection status for at least 3 years. Conclusion: The enteric virome dynamics in gorillas could be potentially associated with the SIVgor status. Virome stability studies clearly provide better markers of pathogenic infection progression than bacteriomes. Further studies are still needed to better understand the influence of SIV pathogenesis on infected gorilla populations in the wild, and to associate deeper taxa (virus genera and species) to SIV status in these animals. 219 EARLY HIV-1 DETECTION IN HUMANIZED MICE Hang Su , Prasanta Dash, Larisa Y. Poluektova, Santhi Gorantla, Howard E. Gendelman Univ of Nebraska, Omaha, NE, USA Background: HIV has limited species tropismwith viral growth only observed in humans and chimpanzees. This constrains pathobiological, prevention and therapeutic studies to primates. In order to overcome restrictions for the use of small animal models, CD34+ hematopoietic stem cells(HSC) were transplanted into immune-deficient NOD.Cg- NOD. Cg-Prkdcscid Il2rgtm1Wjl/SzJ(NSG) mice. They have served as a platform for studies of HIV-virology, pathogenesis, treatment, prevention and eradication. HIV reservoirs that persist in patients even under suppressive antiretroviral therapy(ART) are major obstacles to viral “cure”. As HIV reservoirs are established early in infection and can be restricted by ART, accurate timing for therapeutic intervention is essential to elicit “best” clinical outcomes. Studies of ART timing are thus essential to best understand HIV latency and provide guidance for viral eradication strategies. Methods: Twenty humanized-NSG mice at 18weeks of age were injected intraperitoneally with HIV-1ADA at 104-tissue culture infective dose50 with 5 uninfected mice as controls. Infected animals were randomly separated into 4 groups and sacrificed at day 3, 5, 7 and 14 after infection. Blood and tissues(spleen liver lung gut kidney and brain) were harvested for immunohistochemistry(HLA-DR and HIV-1p24) and semi-nested real-time PCR tests for HIV-1 DNA and RNA. Results: Peripheral CD4+ T cell were reduced by 20% after HIV-1 infection in all the groups. IHC-tests showed higher HIV-1p24+ cells at 14 days as compared to other time points. Plasma viral-loads(VL) were between 103 to 105 copies/ml in all 14-day infected animals. VL was detectable in 2 of 5 animals measured by COBAS-Ampliprep HIV-1 detection kit(detection limit<20copies/ml) from days 3, 5 and 7 groups. HIV-1RNA and DNA in all day-14 animals were measured by semi-nested PCR from tissues(spleen lung and gut, 106–108copies/106hCD45+cells), whereas detected(~106 copies) in 2/5 animals in days 3, 5 and 7 tissues(spleen lung liver and gut, detection limit<10copies). HIV-1 was not detected in brain samples tested. We observed a positive correlation between plasma VL and tissue HIV by semi-nested q-PCR in all infected animal groups. Conclusion: HIV-1 can be seen in CD34+ humanized-NSG mice as early as 3 days after infection at low levels. The timing for fully HIV detection in plasma and tissues in this model is 14 days of infection, at which time any therapy can be started and tissue reservoirs in myeloid and other compartments are established. 220 AN ENGINEERED RNA VIRUS CAUSES ACUTE AIDS IN HUMAN CD4,CCR5 TRANSGENIC MICE Rachel A. Liberatore, Emily J. Mastrocola, Elena Cassella, Jessie Willen, Fabian Schmidt, Dennis Voronin, Trinity Zang, Theodora Hatziioannou, Paul Bieniasz The Rockefeller Univ, New York, NY, USA Background: The rarity of the development of broadly neutralizing antibodies (bNAbs) combined with the lack of an inexpensive, tractable animal model has hampered the pursuit of an effective HIV-1 vaccine. In order to model interactions between HIV-1 Env and its receptors in an in vivo setting, we have generated replication competent, recombinant RNA viruses (RhIV) expressing CCR5-tropic HIV-1 envelopes frommultiple clades, including transmitted founder viruses. These RhIVs are able to infect transgenic mice bearing CD4-cell restricted expression of human CD4 and CCR5. This model, by focusing entirely on Env-receptor interactions, represents a system in which strategies to block these interactions can be rapidly and rigorously tested. Additionally, these RhIVs are capable of eliciting anti-HIV-1 Env Ab responses and therefore may also be pursued as possible vaccine candidates. Mirela D’arc 1 , Carolina Furtado 2 , Juliana D. Siqueira 2 , Ahidjo Ayouba 3 , Martine Peeters 3 , Marcelo A. Soares 2 1 Univ Fed do Rio de Janeiro, Rio de Janeiro, Brazil, 2 Inst Nacional de Câncer, Rio de Janeiro, Brazil, 3 IRD, Montpellier, France

Poster and Themed Discussion Abstracts

85

CROI 2017