Longitudinal Analysis of Cerebrospinal Fluid and Plasma HIV-1 Envelope Sequences Isolated From a Single Donor with HIV Asymptomatic Neurocognitive Impairment.

OBJECTIVE: Combined antiretroviral treatment (cART) has changed the clinical presentation of HIV-associated neurocognitive disorders (HAND) to that of the milder forms of the disease. Asymptomatic neurocognitive impairment (ANI) is now more prevalent and is associated with increased morbidity and mortality risk in HIV-1-infected people. HIV-1 envelope (env) genetic heterogeneity has been detected within the central nervous system (CNS) of individuals with ANI. Changes within env determine co-receptor use, cellular tropism, and neuropathogenesis. We hypothesize that compartmental changes are associated with HIV-1 env C2V4 during ANI and sought to analyze paired HIV-1 env sequences from plasma and cerebrospinal fluid (CSF) of a female subject undergoing long-term cART. METHODS: Paired plasma and CSF samples were collected at 12-month intervals and HIV-1 env C2V4 was cloned and sequenced. RESULTS: Phylogenetic analysis of paired samples consistently showed genetic variants unique to the CSF. Phenotypic prediction showed CCR5 (R5) variants for all CSF-derived sequences and showed minor X4 variants (or dual-tropic) in the plasma at later time points. Viral compartmentalization was evident throughout the study, suggesting that the occurrence of distinctive env strains may contribute to the neuropathogenesis of HAND. CONCLUSIONS: Our study provides new insights about the genetic characteristics within the C2V4 of HIV-1 env that persist after long-term cART and during the course of persistent ANI.

Depression Correlates with Increased Plasma Levels of Inflammatory Cytokines and a Dysregulated Oxidant/Antioxidant Balance in HIV-1-Infected Subjects Undergoing Antiretroviral Therapy.

OBJECTIVE: Depression is the most common psychiatric diagnosis in the HIV/AIDS population and represents a risk factor for disease progression. Since HIV-1 infection is characterized by immunologic and metabolic disturbances, we want to study the effects of depression on different components related to pro-inflammatory and oxidative stress markers. We hypothesize that depression will lead to increased pro-inflammatory cytokine levels and altered antioxidant/oxidant balance. METHODS: We included males and females who were ≥21 years of age, whose HIV-1 sero-status was confirmed by Western Blot, and who were currently undergoing antiretroviral treatment. Patients completed the participation consent form, a socio-demographic survey, and the Patient Health Questionnaire-9 (PHQ-9) for depression assessment. We isolated the plasma from participants' blood samples for viral load analysis (RT-PCR), T-cell counts (flow cytometry), and hematological parameters. A cytokine magnetic bead panel was used to measure interleukin-15 (IL-15), interferon gamma-induced protein 10 (IP-10), IL-12 and granulocyte colony-stimulating factor (G-CSF) levels. We also performed assays to determine the antioxidant activity of superoxide dismutase (SOD) and catalase and to measure the lipid peroxidation levels using malondialdehyde (MDA) and 8-isoprostane assays. Statistical comparisons and correlations at 5% level of significance were determined. RESULTS: Our results show that subjects with mild/moderate to severe depression as assessed by PHQ-9 had a significantly decreased adherence to anti-retroviral treatment. Subjects with depression also had significantly lower levels of white blood cells (WBC) and platelets (PLT) than did the non-depressed group. The HIV+ subjects with depression had increased levels of IL-15, IP-10, IL-12 p40/p70 and G-CSF compared to their non-depressed counterparts. The latter had increased MDA and 8-isoprostane levels. CONCLUSIONS: Our results suggest that HIV+ subjects with depressive symptoms have higher levels of inflammation and altered oxidant/antioxidant balance. Although the groups were small, this study strengthens the hypothesis that alterations in cytokines are associated with the mechanisms underlying depression symptoms.

Identification and molecular characterization of SIV Vpr R50G mutation associated with long term survival in SIV-infected morphine dependent and control macaques.

Viral protein R (Vpr) is an accessory protein of HIV and SIV involved in the pathogenesis of viral infection. In this study, we monitored SIV evolution in the central nervous system and other organs from morphine-dependent and control animals by sequencing vpr in an attempt to understand the relationship between drug abuse, disease progression, and compartmentalization of viral evolution. Animals in the morphine group developed accelerated disease and died within twenty weeks post-infection. A unique mutation, R50G, was identified in the macaques that survived regardless of morphine exposure. Functional studies revealed that the R50G mutation exhibited altered cellular localization and decreased the expression levels of both IL-6 and IL-8. Our results, therefore, suggest that sequence changes within the SIV/17E-Fr vpr occur regardless of drug abuse but correlate with survival, and that they alter disease progression rates by affecting Vpr functions.

Short communication: Lack of immune response in rapid progressor morphine-dependent and SIV/SHIV-infected rhesus macaques is correlated with downregulation of TH1 cytokines.

Our previous studies have shown two distinct disease patterns (rapid and normal onset of clinical symptoms) in morphine-dependent SHIV/SIV-inoculated rhesus macaques. We have also shown that control as well as 50% of morphine-dependent macaques (normal progressor) developed humoral and cellular immune responses whereas the other half of the morphine-dependent macaques (rapid progressor) did not develop antiviral immune responses after infection with SIV/SHIV. In the present study, we analyzed the association between cytokine production, immune response, and disease progression. To study the immunological effects of morphine at cytokine levels in the context of a lentiviral infection, we inoculated rhesus macaques with a mixture of SHIV(KU-18), SHIV(89.6)P, and SIV/17E-Fr. These animals were followed for a period of 56 weeks for cytokine level production in plasma. Drug-dependent rapid disease progressors exhibited an increase in IL-18 and IL-1Ra and a decrease in IL-12 levels in the plasma. Morphine-dependent normal progressors and control macaques exhibited an increase in both IL-18 and IL-12, whereas IL-Ra levels remained constant throughout the observation period. These results suggest that rapid disease progression in relation to morphine dependency may be the result of an altered cytokine profile.

Accelerated evolution of SIV env within the cerebral compartment in the setting of morphine-dependent rapid disease progression.

Human immunodeficiency virus-1 (HIV-1) and simian immunodeficiency virus (SIV) have been shown to compartmentalize within various tissues, including the brain. However, the evolution of viral quasispecies in the setting of drug abuse has not been characterized. The goal of this study was to examine viral evolution in the cerebral compartment of morphine-dependent and control macaques to determine its role in rapid disease progression. To address this issue, we analyzed the envelope (env) gene from proviral DNA in our SIV/SHIV macaque model of morphine dependence and AIDS. Analyses of proviral DNA revealed a direct correlation between total genetic changes and survival time. However, the rate of evolution during disease progression was higher in morphine-dependent and rapid-progressor macaques than was the rate of evolution in the control animals. This study provides additional insight into SIV envelope variation in the CNS of morphine-dependent macaques and genotypes that may have evolved in the brain and contributed to disease progression.