Changes in the plasma proteome follows chronic opiate administration in simian immunodeficiency virus infected rhesus macaques.

BACKGROUND: Substantive plasma proteomic changes follow lentiviral infection and disease pathobiology. We posit that such protein alterations are modified during drug abuse, further serving to affect the disease. To this end, we investigated the effect of opiate administration on the plasma proteome of Indian-strain rhesus monkeys infected with simian immunodeficiency virus (SIV) strain smm9. METHODS: Whole blood was collected at 7 weeks prior to and 1.4 and 49 weeks after viral infection. Viral load, CD4(+) T cell subsets, and plasma protein content were measured from monkeys that did or did not receive continuous opiate administrations. The plasma proteome was identified and quantified by isobaric tags for relative and absolute quantitation labeling (iTRAQ) and mass spectrometry. RESULTS: While substantive changes in plasma proteins were seen during SIV infection, the addition of opiates led to suppression of these changes as well as increased variance of the proteome. These changes demonstrate that opiates induce broad but variant immune suppression in SIV-infected monkeys. CONCLUSION: The broad suppressive changes seen in plasma of SIV-infected monkeys likely reflect reduced multisystem immune homeostatic responses induced by opiates. Such occur as a consequence of complex cell-to-cell interactions operative between the virus and the host. We conclude that such changes in plasma proteomic profiling may be underappreciated and as such supports the need for improved clinical definitions.

Neurometabolite abnormalities in simian immunodeficiency virus-infected macaques with chronic morphine administration.

Opiate abuse increases the risk for human immunodeficiency virus (HIV) infection, while both opiates and HIV may impact the immune and nervous systems. To model potential interactions between opiate drugs and HIV on the brain, neurometabolite levels were evaluated in simian immunodeficiency virus (SIV)-infected macaques with or without chronic morphine administration. Over the course of the study, 58% of these SIV-infected animals progressed to acquired immune deficiency syndrome (AIDS). Brain extracts from four brain regions were evaluated with proton magnetic resonance spectroscopy. Animals with AIDS had lower N-acetyl-aspartate in all four brain regions (p ≤ 0.05) as well as lower frontal gray matter total creatine (p= 0.03), lower frontal white matter (p= 0.003) and caudate (p = 0.002) glutamate, and higher frontal white matter myo-inositol (p= 0.05) than the healthier non-AIDS macaques. Morphine-dependent animals had higher levels of myo-inositol in the putamen (p = 0.003), especially those with AIDS. In the animals with AIDS, those with morphine dependence had higher total creatine in the frontal white matter (p= 0.04) than those treated with saline, which in turn had lower creatine than saline-injected animals without AIDS (p = 0.04), leading to an interaction between the effects of morphine and AIDS on total creatine in this brain region (ANOVA p = 0.02). The majority of these brain metabolites correlated with viral counts indicating more severe metabolite abnormalities in animals with higher viral loads or set points. Collectively, these findings suggest that chronic morphine may protect against the neurotoxic effect of AIDS and reinforce the importance of maintaining a low viral load in AIDS.

Plasma proteomic analysis of simian immunodeficiency virus infection of rhesus macaques.

Lentiviral replication in its target cells affects a delicate balance between cellular cofactors required for virus propagation and immunoregulation for host defense. To better elucidate cellular proteins linked to viral infection, we tested plasma from rhesus macaques infected with the simian immunodeficiency viral strain SIVsmm9, prior to, 10 days (acute), and 49 weeks (chronic) after viral infection. Changes in plasma protein content were measured by quantitative mass spectrometry by isobaric tags for absolute and relative quantitation (iTRAQ) methods. An 81 and 232% increase in SERPINA1 was seen during acute and chronic infection, respectively. Interestingly, gelsolin, vitamin D binding protein and histidine rich glycoprotein were decreased by 45% in acute conditions but returned to baseline during chronic infection. When compared to uninfected controls, a 48-103% increase in leucine rich alpha 2-glycoprotein, vitronectin, and ceruloplasmin was observed during chronic viral infection. Observed changes in plasma proteins expression likely represent a compensatory host response to persistent viral infection.

Probable deceleration of progression of Simian AIDS affected by opiate dependency: studies with a rhesus macaque/SIVsmm9 model.

OBJECTIVES: To determine effects of opiate dependency on development of simian AIDS. DESIGN: Assessments of viral, immune, and clinicopathological status were conducted on rhesus macaques before and after establishment of opiate dependency and Simian Immunodeficiency Virus, sooty mangabey, strain-9 (SIVsmm9) infection. Controls received saline. METHODS: Blood was collected at baseline, before opiate dependencies, and viral infections were established and then after SIVsmm9 infection, longitudinally, through 216 weeks. Plasma viral titers were assessed using the branched chain DNA assay and CD4 and CD8 counts via cytofluorometry. Clinicopathological assessments of AIDS were founded on Centers for Disease Control and Prevention and other selected criteria. RESULTS: AIDS progression rates seemed to be decelerated and survival times increased by opiate dependency. Mean viral titers were unaffected by opiate exposure. Opiate-dependent monkeys that evidenced high initial viral titers survived significantly longer than controls. Several opiate-dependent monkeys maintained high viral titers for atypically extended durations. Several (5/19) opiate-dependent monkeys died or were removed early from the study due to "non-AIDS" causes. CONCLUSIONS: Long-term opiate dependency seemed to decelerate the rate of progression to AIDS in the SIVsmm9 monkey model. This effect was most evident in monkeys with high initial viral titers/set points. "Non-AIDS" morbidities and mortalities were noted as potential confounds of epidemiological assessments of the role of opiates in HIV/AIDS.

Tissue-specific reduction in DC-SIGN expression correlates with progression of pathogenic simian immunodeficiency virus infection.

Studies were undertaken to determine whether previously described reductions in splenic DC-SIGN expression in simian acquired immune deficiency syndrome (AIDS) are limited to pathogenic simian immunodeficiency virus (SIV) infection. DC-SIGN expression was evaluated by immunohistochemistry in lymphoid tissues from AIDS-susceptible Asian macaque monkeys as compared with AIDS-resistant sooty mangabey monkeys in the presence and absence of SIV infection. The phenotype of DC-SIGN+ cells in susceptible and resistant species was identical and most consistent with macrophage identity. Significantly lower levels of DC-SIGN expression were identified in spleen, mesenteric lymph node, and bone marrow of macaques with AIDS (P<0.05). Reduced levels of splenic DC-SIGN correlated significantly with CD4T cell depletion in long-term pathogenic infection of macaques (P<0.01), whereas SIV-infected mangabeys retained high levels of DC-SIGN expression in spleen despite persistent infection. Reduced expression of DC-SIGN in spleen specifically characterizes pathogenic forms of SIV infection, correlates with disease progression, and may contribute to SIV pathogenesis.

The role of substance abuse in HIV disease progression: reconciling differences from laboratory and epidemiologic investigations.

From the onset of the HIV/AIDS epidemic, the use of licit and illicit drugs has been investigated for its potential impact on HIV disease progression. Findings from a large number of laboratory-based studies indicate that drug abuse may exacerbate HIV disease progression; however, epidemiological studies have shown mixed results. This article presents a review of findings from both laboratory-based and epidemiologic investigations. In addition, we provide a careful evaluation of methodological strengths and limitations inherent to both study designs in order to provide a more nuanced understanding of how these findings may complement one another.

Modulation by morphine of viral set point in rhesus macaques infected with simian immunodeficiency virus and simian-human immunodeficiency virus.

Six rhesus macaques were adapted to morphine dependence by injecting three doses of morphine (5 mg/kg of body weight) for a total of 20 weeks. These animals along with six control macaques were infected intravenously with mixture of simian-human immunodeficiency virus KU-1B (SHIV(KU-1B)), SHIV(89.6P), and simian immunodeficiency virus 17E-Fr. Levels of circulating CD4(+) T cells and viral loads in the plasma and the cerebrospinal fluid were monitored in these macaques for a period of 12 weeks. Both morphine and control groups showed precipitous loss of CD4(+) T cells. However this loss was more prominent in the morphine group at week 2 (P = 0.04). Again both morphine and control groups showed comparable peak plasma viral load at week 2, but the viral set points were higher in the morphine group than that in the control group. Likewise, the extent of virus replication in the cerebral compartment was more pronounced in the morphine group. These results provide a definitive evidence for a positive correlation between morphine and levels of viral replication.

Multiple ways that drug abuse might influence AIDS progression: clues from a monkey model.

Whether opiates and other drugs of abuse affect AIDS progression has been an unresolved issue for two decades. Credible evidence has suggested that opiates may exacerbate, retard or have 'no effect' on progression of AIDS. Differences may exist in AIDS-progression outcomes after opiate exposures that relate to neural versus, strictly, somatic AIDS; but it is also likely that conditional variables inherent to drug dependency and the nature of the infectious agents involved allow for differing outcomes. Data from epidemiological studies, and from in vitro and basic immunological studies regarding opiate effects on AIDS progression must be interpreted in light of the conditionality of opiate effects. Caution is sounded, also, about interpreting common depressive immunological effects of opiates as indicators of influence over AIDS progression. Current evidence from the monkey model of AIDS indicates that opiates can, under certain defined conditions, retard AIDS progression. The clinical relevance of these data remains to be defined. Importantly, such data imply that opioids and the endogenous opioid system may represent therapeutic tools and targets for altering AIDS progression.

RXR-induced TNF-alpha suppression is reversed by morphine in activated U937 cells.

Deficiency in vitamin A has been associated with adverse clinical outcomes in drug users with HIV-1 infection. Retinoids have been demonstrated to suppress proinflammatory cytokine production by immune cells in vitro. These effects are induced by ligand-mediated activation of the retinoid receptors--retinoic acid receptor (RAR) and retinoid X receptor (RXR). In these studies, the effects of all-trans-retinoid acid (ATRA, a RAR agonist), 9-cis-retinoic acid (9cis RA; RAR and RXR agonist), LG101305 (RXR agonist), LG100815 (RAR antagonist) and LG101208 (RXR antagonist) on TNF-alpha production by phytohemagglutanin-activated U937 cells and the modulation of these effects by morphine were examined. TNF-alpha production was suppressed in all cultures exposed to retinoid agonist and antagonist agents. For cells exposed to RXR agonists or RAR antagonist, incubation with morphine resulted in the reversal of TNF-alpha suppression and this effect was inhibited by naloxone. These data suggest that interactions between RXR and morphine are involved in the immune effects of retinoids on TNF-alpha production by activated U937 cells. Such information may be important for understanding interactions between drugs of abuse and immune function in individuals with chronic proinflammatory states such as HIV-1 infection.

Does reduced DNA repair capacity play a role in HIV infection and progression in the lymphocytes of opiate addicts?

Opiate use in vivo and in vitro reduces the ability of human peripheral lymphocytes to repair DNA damage caused by both the physical and chemical mutagens that produce single-strand adducts. This decrease in repair leads to increased genetic damage to the individual cell as measured by cytogenetic damage, including sister chromatid exchanges and formation of micronuclei. The expected consequences of this increase in damage can also be established by increases in host cell mutation rate and rate of apoptosis. The effect of this increase in genetic damage can be expected to have significant consequences for HIV-1 or simian immunodeficiency virus infecting those lymphocytes. For example, DNA damaging agents have long been known to induce lentiviral growth and propagation, and this has been found to be true for HIV-1 following ultraviolet light treatment of lymphocytes. However, to date, no one has fully explored the consequences of increased host mutation rate on HIV growth and maintenance. Recent reports have demonstrated the role of viral mutation in such key physiologic processes as resistance to highly active antiretroviral therapy (HAART). Beyond the effects of random mutations in the viral genome, specific mutations in the HIV-1 transcriptase and protease lead to increased accumulation of mutant viruses and the gradual failure of HAART. It therefore remains to be tested whether changes in host cell mutation rate will also predict changes in susceptibility to drug therapy. This also leads to questions about whether the higher rate of viral mutation in HIV-infected drug addicts might be the basis for higher rates of neuroAIDS in this population. It would be attractive to speculate that the increase in the heterogeneity of the virus in addicts produces mutants with a greater capacity to attack neuronal tissue and a high affinity to replicate there.