CROI 2018 Abstract eBook

Abstract eBook

Oral Abstracts

71 NO RESIDUAL VIRUS REPLICATION IN A RANDOMISED TRIAL OF DOLUTEGRAVIR INTENSIFICATION

Thomas A. Rasmussen 1 , James McMahon 2 , J. Judy Chang 3 , Jennifer Audsley 3 , Ajantha Solomon 3 , Surekha Tennakoon 3 , Ashanti Dantanarayana 3 , Tim Spelman 3 , Tina Schmidt 2 , Stephen J. Kent 3 , Vincent Morcilla 4 , Sarah Palmer 4 , Julian Elliott 2 , Sharon R. Lewin 3 1 Aarhus University Hospital, Aarhus, Denmark, 2 Alfred Hospital, Melbourne, VIC, Australia, 3 Doherty Institute for Infection and Immunity, Melbourne, VIC, Australia, 4 University of Sydney, Westmead, NSW, Australia Background: Whether residual virus replication (RVR) persists in HIV-infected individuals on suppressive antiretroviral therapy (ART) remains controversial. One strategy used to demonstrate RVR is to intensify ART with an integrase inhibitor and measure an early increase in 2-long terminal repeat (2-LTR) circles. Two previous studies with raltegravir demonstrated RVR in a subset of individuals on ART. Here we investigated the effects of dolutegravir. Methods: In a randomised, placebo-controlled, double-blinded clinical trial, HIV-infected adults with virological suppression for >3 years were randomly assigned 1:1 to dolutegravir 50 mg or placebo daily for 56 days in addition to background ART. The primary outcome measure was the level of 2-LTR circles in CD4+ T cells at day 7. Cell-associated unspliced (CA-US) HIV RNA, total and integrated HIV DNA, and plasma HIV RNA using a single copy assay (SCA) were quantified by real-time PCR; T cell expression of HLA-DR, CD38 and PD-1 by flow cytometry, and plasma levels of interleukin-6 (IL-6), high-sensitivity C-reactive protein (hsCRP), d-dimer and soluble CD14 (sCD14) by ELISA. We used repeated- measures analysis of variance (ANOVA) as the protocol-defined primary analysis. Student’s t-test or rank sum test, were used to compare changes from baseline to specific time points across study arms. Results: We enrolled 40 HIV-infected individuals; 21 were allocated to dolutegravir and 19 to placebo with 14 and 11% receiving a protease-inhibitor based ART regimen respectively. All participants completed the study. There was no significant difference in the primary endpoint, 2-LTR circles in peripheral blood CD4+ T cells, as assessed by repeated-measures ANOVA over 7 days (p=0.17) or any other time point (Figure). Median (IQR) 2-LTR circles fold- change from baseline to day 7 was -0.17 (-0.90 to 0.90) in the dolutegravir and -0.26 (-1.00 to 1.17) in the placebo groups. We found no consistent difference in the levels of CA US HIV-RNA, total and integrated HIV DNA (Figure), SCA, T cell activation markers or plasma levels of sCD14, d-dimer, IL-6 or hs-CRP. PD-1 expression in CD4+ T cells declined slightly after 56 days in placebo recipients compared to dolutegravir (p=0.03). Conclusion: In a randomised, double-blinded, placebo-controlled trial of dolutegravir intensification, there was no evidence of RVR on ART.

Oral Abstracts

70 NO EVIDENCE FOR ONGOING HIV REPLICATION IN LYMPH NODES DURING SUPPRESSIVE ART William R. McManus 1 , Michael J. Bale 1 , Jonathan Spindler 1 , Ann Wiegand 1 , Andrew Musick 1 , Xiaolin Wu 2 , David Wells 2 , Stephen H. Hughes 1 , Brandon Keele 2 , Rebecca Hoh 3 , Jeffery Milush 3 , John M. Coffin 4 , John W. Mellors 5 , Steven G. Deeks 3 , Mary F. Kearney 1 1 NIH, Frederick, MD, USA, 2 Leidos Biomedical Research, Inc, Frederick, MD, USA, 3 University of California San Francisco, San Francisco, CA, USA, 4 Tufts University, Boston, MA, USA, 5 University of Pittsburgh, Pittsburgh, PA, USA Background: Lymph nodes have been implicated as potential sanctuary sites of ongoing HIV replication during ART due to inadequate drug penetration. To investigate this possibility, we characterized HIV proviral populations, their levels of expression, and their sites of integration in paired lymph node (LNMC) and peripheral blood (PBMC) samples collected after long-term ART. Methods: PBMC and LNMC were obtained from five donors: four had <40 cps/ml on ART for 4.3-12.9 years and one was ART-naïve. Pre-ART samples were obtained for three of the treated patients. Longitudinal on-ART LNMC were obtained from two patients one year apart, including sampling from two different inguinal nodes in both individuals. Proviral populations and expression were characterized by cell associated RNA- and DNA- single genome sequencing of p6-PR-RT. Proviral sequences were compared phylogenetically and by testing for panmixia. Infected cell clones were identified by integration sites assay (ISA) in PBMC and LNMC from one donor. Results: Comparisons of the proviral sequences on ART in PBMC (n=176) and LNMC (n=234) showed no increase in branch length, diversity, or divergence from pre-ART plasma or PBMC due to ongoing viral replication in either location. A test for panmixia of proviral sequences in PBMC and LNMC and across two separate lymph nodes sampled at the same time point showed no evidence for compartmentalization (probability of panmixia p>0.3). Proviruses with identical sequences were found in LNMC and PBMC and were transcriptionally active at both sites, although a greater fraction of infected cells in LNMC was expressing HIV RNA than in PBMC (13% vs. 6%). High-expressing cells (>20 HIV RNA copies/cell) were observed in samples obtained prior to but not during ART, with the exception of one LNMC. In one patient, forty clones of infected cells were identified by ISA. There were no differences in the locations of these clones in PBMC vs. LNMC (p=0.8). populations of infected cells were well-mixed. There was no evidence of tissue compartmentalization. There was also no evidence for divergence from pre-ART populations in PBMC or in LNMC whether ART was initiated in acute or chronic infection, which is not consistent with the HIV reservoir being maintained by ongoing cycles of viral replication in either PBMC or LNMC during suppressive ART. Conclusion: Comparison of proviral populations, including clones of infected cells, and their expression in LNMC and PBMC, showed that

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CROI 2018