CROI 2020 Abstract eBook

Abstract eBook

Poster Abstracts

Background: CSF is a possible compartmentalized HIV reservoir though the cells involved and their level of HIV-1 transcriptional activity remain obscure. We used a novel highly sensitive assay of HIV-1 RNA/DNA and flow cytometry to study both CSF cells and PBMC. Methods: We studied 10 HIV+ subjects (2 with current HAND) on cART with both plasma and CSF HIV RNA (Roche) <50 copies/ml.DNA and RNA were extracted from paired samples of CSF (13-20 ml) and PBMC. Cell-associated HIV-1 transcriptional activity and HIV-1 DNA levels were determined by a newly described quantitative piCode End-Point PCR assay, based on the extremely sensitive piCode MicroDiscs platform (>27-fold sensitivity of real-time PCR; Suzuki et al J AIDS HIV Treat. 2019; 1(2):69). It detects transcripts of HIV LTR, including unspliced RNA (gag/pol), incompletely spliced RNA (tat, vpr, vpu), and multiply spliced RNA (rev, nef), with a sensitivity of 3 infected cells/10 6 cells. Immunological profiles of CSF cells and PBMC were determined by 18-colour flow cytometry and compared by Wilcoxon signed rank test. MR spectroscopy (MRS) evaluated the frontal white matter (FWM), posterior cingulate cortex (PCC), and caudate nucleus. Results: 9/10 patients’ CSF had high levels of cell-associated HIV-1 RNA transcriptional activity (median 4711 copies per 10 6 cells, vs 270 in PBMC; p=0.004). 8/10 patients had HIV-1 DNA in CSF cells (median 1314 copies per 10 6 cells vs 752 in PBMC; p=0.09). Higher HIV-1 RNA in CSF cells was correlated with lower N-acetyl aspartate in FWM (r=-0.78; p=0.038) and PCC (r=-0.76; p=0.012). 95% of CSF cells were T cells, of which 95%were memory CD4 and CD8 T cells (median counts of 8,818 and 7,503 cells, respectively). 2.8% of CSF cells were CD14+CD16+monocytes, 1.7%were NK cells and 0.4 %were B cells. CSF CD4 T cells consisted of 75% CXCR3+ CD49d+ integrinß7- cells (vs 15% of CD4 in PBMC); 48% CCR5+ (vs 16% in PBMC); and 18% expressing CD38 and/or HLA-DR activation markers (vs 7% in PBMC). Conclusion: CSF cellular HIV-1 LTR transcriptional activity is compartmentalised and its biological significance is strongly indicated by the MRS correlations. The cellular origin is likely the dominant CXCR3+ CD49d+ integrinß7- non-gut homing memory CD4+T cells; monocytes may be less important. Transcriptional products eg tat (vs whole virus) are likely neuropathogenetically significant. These data support HIV-1 transcription inhibitor development. 430 PLASMA RAMs IN REVERSE TRANSCRIPTASE GENE ASSOCIATE WITH CSF HIV-1 ESCAPE Mattia Trunfio 1 , Anna Celotti 2 , Francesca Bai 3 , Luigi Celani 4 , Emanuele Focà 2 , Antonella D'Arminio Monforte 3 , Stefano Bonora 1 , Gabriella D'Ettorre 4 , Giovanni Di Perri 1 , Andrea Calcagno 1 1 University of Torin, Torino, Italy, 2 University of Brescia, Brescia, Italy, 3 University of Milan, Milan, Italy, 4 Sapienza University of Rome, Rome, Italy Background: Several risk factors for cerebrospinal fluid HIV-1 escape (CSFE) have been reported: length of HIV infection, cART interruptions, low CD4 nadir/ CPE score, persistent low-level viremia and the use of ABC+3TC, boosted PIs or unboosted ATZ. We sought to assess whether the presence of previous plasma RAMs may be a determinant behind the reported risk association between CSFE and ARVs class composing cART. Methods: Retrospective cross-sectional study on HIV+ adult patients on cART undergoing lumbar puncture (LP) for any reason (2007-July 2019) at 4 Italian hospitals (Brescia, Torino, Roma, Milano). Inclusion criteria: being on cART for at least 6 months, available coupled plasma and CSF HIV-RNA measurements, available historical cumulative plasma genotypic resistance testing (HGRT) for reverse transcriptase (RT) and protease (PI) genes. Exclusion criteria: secondary CSFE. CSFE was defined as any measurable CSF HIV-RNA coupled with a plasma HIV-RNA <50 copies/mL and any difference ≥0.5 Log 10 between CSF and plasma HIV-RNA when the latter was detectable Results: 197 patients were enrolled: 50 years (43-54), current and nadir CD4 count 312 (115-560) and 82 (24-200) cells/μL; median length of cART treatment

54 months (17-171). 126 patients (63.9%) had plasma HIV-RNA<50 cp/mL and 28 (14.2%) showed CSFE. The main reasons for LP were diagnostic assessment in diseases without eventually CNS involvement (25.4%), HIV-associated neurocognitive disorders (28.4%), CNS infections (19.8%) and research purposes (16.2%). CSFE was not associated with PIs use in the whole cohort (16.6% vs 8.6%, p.14) nor in any subgroup identified by cART type (3 different-classes-, 3 drugs-NRTIs- and ≥4 drugs-based cART). Instead, PIs use was more common in patients with a positive HGRT for RAMs in RT (44.9% vs 29.3%, OR 2.0 [1.1-3.8], p.04). Having a cumulative HGRT positive for RAMs in RT associated with a higher risk of CSFE (21.5% vs 9.3%, OR 2.7 [1.2-6.0], p.01), while no such an association was observed for RAMs in PI (17.4% vs 13.8%). Interestingly, at the CSFE diagnosis patients showed higher proportion of positive CSF RAMs in RT compared to patients without CSFE with available CSF GRT (55.6% vs 19.0%, p.04). At multivariable analysis, only RAMS in RT and CD4 nadir were independent predictors of CSFE (tab.1) Conclusion: In this cohort, CSFE prevalence was slightly higher than what reported in recent studies. Besides low CD4 nadir, the positivity of HGRT for plasma RAMs in the RT gene and not the use of PIs per sé was an independent predictor of CSFE 431 PLASMA AND CSF SCD30 DYNAMICS BEFORE AND AFTER ART INITIATED IN ACUTE HIV Michael J. Peluso 1 , Bonnie Slike 2 , Phillip Chan 3 , Carlo Sacdalan 3 , Siriwat Akapirat 4 , Linda Jagodzinski 2 , Victor Valcour 1 , Nittaya Phanuphak 3 , Eugène Kroon 3 , Jintanat Ananworanich 2 , Sandhya Vasan 2 , Timothy J. Henrich 1 , Serena S. Spudich 5 , Shelly J. Krebs 2 , for the RV254/SEARCH 010 Study Team 1 University of California San Francisco, San Francisco, CA, USA, 2 US Military HIV Research Program, Silver Spring, MD, USA, 3 Thai Red Cross AIDS Research Center, Bangkok, Thailand, 4 Armed Forces Research Institute of Medical Sciences in Bangkok, Bangkok, Thailand, 5 Yale University, New Haven, CT, USA Background: Soluble CD30 (sCD30) is a potential marker of persistent immune activation and/or viral persistence in people living with HIV (PLWH). Surface CD30 co-localizes with HIV RNA and DNA in CD4+ T cells from blood and gut tissue and depletion of cells expressing CD30 reduces the total amount of HIV-1 DNA detected. Evalution of sCD30 in cerebrospinal fluid (CSF) compared to blood before and after antiretroviral therapy (ART) in acute HIV (AHI) may be valuable in understanding HIV neuropathogenesis. Methods: We measured pre-ART sCD30 in plasma (n=117; 71 Fiebig 1-2, 46 Fiebig 3-5) and CSF (n=71; 39 Fiebig 1-2, 32 Fiebig 3-5) in the Thai RV254/ SEARCH010 AHI cohort and examined correlations with HIV disease parameters and inflammatory biomarkers. A subset had sCD30 levels measured at 48 and 96 weeks on ART in plasma (n=109 and n=56, respectively) and CSF (n=40 and n=20, respectively). We used non-parametric tests to compare sCD30 levels between AHI participants and HIV-uninfected Thais and to assess relationships between covariates. We used mixed effects models to examine changes within individuals over time. Results: The median age was 26.5 years (IQR 23-31), pre-ART CD4 count 381 (276-519), and estimated duration of infection 18 (15-23) days. The sample was 96%male. Compared with controls, pre-ART sCD30 levels were elevated in plasma (984 vs 374 pg/mL, p<0.001) and CSF (165 vs 131 pg/mL, p=0.01). Pre-ART plasma sCD30 levels correlated with plasma HIV RNA (r=0.44, p<0.001) and CD4/CD8 ratio (r=-0.5, p<0.001) as well as plasma neopterin (r=0.29, p=0.01), sCD163 (r=0.26, p=0.03), and IP-10 (r=0.28, p=0.02). Pre-ART CSF sCD30 correlated with CSF HIV RNA (r=0.24, p=0.03) as well as CSF sCD14 (r=0.44, p=0.001), sCD163 (r=0.46, p<0.001), and IP-10 (r=0.26, p=0.02). Plasma and CSF sCD30 did not correlate. In longitudinal analyses, sCD30 levels in both compartments declined at 48 and 96 weeks. This decline was more substantial in plasma (-1.9-fold change, p<0.001) than CSF (-1.14-fold change, p=0.0015). Conclusion: In untreated AHI, sCD30 is elevated in plasma and CSF and correlates with markers of HIV disease activity and inflammation. With ART initiation in AHI, sCD30 levels decline in both compartments; this is distinct from

Poster Abstracts

CROI 2020 150