CROI 2017 Abstract e-Book

Abstract eBook

Poster and Themed Discussion Abstracts

368 THE EFFECT OF CRYPTOCOCCAL MENINGITIS ON THE HIV RESERVOIR IN THE CNS Maura Manion 1 , Camille Lange 2 , Kathy Huppler Hullsiek 3 , Brandon Keele 4 , Helene Highbarger 5 , David Meya 6 , Frank Maldarelli 4 , David Boulware 3 , Irini Sereti 1 1 NIAID, Bethesda, MD, USA, 2 NIH, Frederick, MD, USA, 3 Univ of Minnesota, Minneapolis, MN, USA, 4 NCI, Frederick, MD, USA, 5 Leidos Biomed, Inc, Frederick, MD, USA, 6 Makerere Univ, Kampala, Uganda Background: Opportunistic central nervous system (CNS) infections are postulated to influence the formation and persistence of a HIV CNS reservoir through trafficking of infected and activated cells. Viral compartmentalization present at initiation of antiretroviral therapy (ART) initiation may affect the establishment of a reservoir. We investigated the role CNS infections have in seeding the HIV reservoir by characterizing viral populations and compartmentalization of virus in blood and cerebrospinal fluid (CSF). Methods: Paired CSF and plasma samples from 73 HIV-infected persons with cryptococcal meningitis were obtained prior to ART during two clinical trials in Uganda. In a subset of patients (N=14), single genome sequences (SGS) of HIV-1 envelope were obtained from paired plasma and CSF samples, aligned (MEGA), and subjected to phylogenetic and population genetic analysis, including geographic subdivision to determine panmixia between populations, and Slatkin-Maddison population migration studies to determine structural shifts between populations. Geno2Pheo algorithmwas used to predict co-receptor utilization. Results: Participants (Table) had median CD4 =21 cells/μL and pre-ART HIV-1 viral load (VL) of 5 log10 RNA copies/mL in plasma and 4.6 log10 copies/mL in CSF. CSF VL was lower by a median of 0.3 (IQR: -0.2, 0.9) copies/mL than in plasma. Median CSF VL was higher by 0.42 log10 copies/mL in those with CSF WBC ≥ 5 than in < 5 cells/μL (p=0.03). Single genome sequences (length 21818nt) were obtained. Geographic subdivision and migration studies demonstrated viral compartmentalization between CSF and plasma in 43% (6/14) patients, 4 of 6 with higher plasma VL than CSF and 2 of 8 with higher CSF VL than plasma. Analysis of predicted co-receptors identified X4 variants in CSF and plasma, including patients with compartmentalization; CSF may have predominantly X4 virus even when plasma has predominantly R5 variants. Conclusion: This is the first description of HIV-1 compartmentalization in the CNS during AIDS-related infections. The difference between HIV VL in plasma and CSF in these persons was less than expected. Persons with greater CSF pleocytosis had higher CSF VL. Of cryptococcal meningitis participants, 40% had distinct HIV populations in CSF and in plasma, with a trend of compartmentalization that varied with CSF VL. This provides insights into the seeding of the HIV reservoir during CNS opportunistic infections.

Poster and Themed Discussion Abstracts

369 HIV CNS COMPARTMENTALIZATION AMONG SUBJECTS WITH HIV-1 SUBTYPES G AND CRF02_AG

Olubusuyi M. Adewumi 1 , Elena Dukhovlinova 2 , Nathan Shehu 3 , Shuntai Zhou 2 , Maxwell Akanbi 3 , Babafemi Taiwo 4 , Adesola Ogunniyi 1 , Kevin Robertson 2 , Ronald Swanstrom 2 1 Univ of Ibadan, Ibadan, Nigeria, 2 Univ of North Carolina at Chapel Hill, Chapel Hill, NC, USA, 3 Jos Univ Teaching Hosp, Jos, Nigeria, 4 Northwestern Univ Feinberg Sch of Med, Chicago, IL, USA, Background: HIV-1 compartmentalization in the central nervous system (CNS) and its contribution to neurological disease are incompletely understood. Previous studies were conducted among subjects infected with subtypes B or C. Little is known about other HIV-1 subtypes. We investigated HIV-1 compartmentalization in the CNS of subjects infected with HIV-1 subtypes G and CRF02_AG, the most prevalent subtypes in parts of sub-Saharan Africa. Methods: A cross-sectional study was conducted among HIV-infected subjects with suspected cryptococcal meningitis at Jos University Teaching Hospital, Nigeria. Paired plasma and cerebrospinal fluid (CSF) samples were collected from 13 (M-6, F-7; age range: 23-45yrs) subjects with a clinical indication for lumbar puncture. Viral RNA extraction, cDNA synthesis, and amplification of the V1/V3 region of env and the full-length HIV-1 env gene were done by PCR techniques using specific sets of primers. Pooled libraries of VI/ V3 amplicons (tagged with Primer ID) and full length HIV env genes were analyzed by deep sequencing and single genome amplification (SGA) assays respectively, followed by phylogenetic analysis to determine evolutionary association. Results: Plasma and CSF viral RNA of the subjects ranged between <400 - 1.0 x106 copies/mL and <400 - 5.1 x106 copies/mL, respectively. Six of the subjects had sufficient levels of virus in the CSF to allow comparison to virus in the blood. All recovered isolates were most closely related to HIV-1 subtypes G or CRF02_AG. Deep sequencing demonstrated HIV-1 compartmentalization in the CNS in subjects A02, A04 and A10, and equilibrated virus populations in the plasma and CSF in subject A13. Further analysis by SGA assay confirmed the findings and showed a diverse level of compartmentalization in the subjects (Figure 1). Determination of the method of entry of the phenotype of the compartmentalized variants is ongoing. Conclusion: We have provided the first documentation of CNS compartmentalization of HIV-1 subtypes G and CRF02_AG. Improved understanding of effects compartmentalization on CNS HIV drug susceptibility, viral reservoir, and neurological disorders in sub-Saharan Africa where these subtypes are dominant is imperative. Extremely high viral RNA levels observed in the CSF of subject A13 may be due to the transfer of infected T cells into the CSF/CNS indicating a possible contribution of the meningitis that is prevalent in HIV-1 patients in the region.

CROI 2017 147