Anti-α4 Antibody Treatment Blocks Virus Traffic to the Brain and Gut Early, and Stabilizes CNS Injury Late in Infection

Four SIV-infected monkeys with high plasma virus and CNS injury were treated with an anti-α4 blocking antibody (natalizumab) once a week for three weeks beginning on 28 days post-infection (late). Infection in the brain and gut were quantified, and neuronal injury in the CNS was assessed by MR spectroscopy, and compared to controls with AIDS and SIV encephalitis. Treatment resulted in stabilization of ongoing neuronal injury (NAA/Cr by 1H MRS), and decreased numbers of monocytes/macrophages and productive infection (SIV p28+, RNA+) in brain and gut. Antibody treatment of six SIV infected monkeys at the time of infection (early) for 3 weeks blocked monocyte/macrophage traffic and infection in the CNS, and significantly decreased leukocyte traffic and infection in the gut. SIV – RNA and p28 was absent in the CNS and the gut. SIV DNA was undetectable in brains of five of six early treated macaques, but proviral DNA in guts of treated and control animals was equivalent. Early treated animals had low-to-no plasma LPS and sCD163. These results support the notion that monocyte/macrophage traffic late in infection drives neuronal injury and maintains CNS viral reservoirs and lesions. Leukocyte traffic early in infection seeds the CNS with virus and contributes to productive infection in the gut. Leukocyte traffic early contributes to gut pathology, bacterial translocation, and activation of innate immunity

Temporal/compartmental changes in viral RNA and neuronal injury in a primate model of NeuroAIDS

Despite the advent of highly active anti-retroviral therapy HIV-associated neurocognitive disorders (HAND) continue to be a significant problem. Furthermore, the precise pathogenesis of this neurodegeneration is still unclear. The objective of this study was to examine the relationship between infection by the simian immunodeficiency virus (SIV) and neuronal injury in the rhesus macaque using in vivo and postmortem sampling techniques. The effect of SIV infection in 23 adult rhesus macaques was investigated using an accelerated NeuroAIDS model. Disease progression was modulated either with combination anti-retroviral therapy (cART, 4 animals) or minocycline (7 animals). Twelve animals remained untreated. Viral loads were monitored in the blood and cerebral spinal fluid, as were levels of activated monocytes in the blood. Neuronal injury was monitored in vivo using magnetic resonance spectroscopy. Viral RNA was quantified in brain tissue of each animal postmortem using reverse transcription polymerase chain reaction (RT-PCR), and neuronal injury was assessed by immunohistochemistry. Without treatment, viral RNA in plasma, cerebral spinal fluid, and brain tissue appears to reach a plateau. Neuronal injury was highly correlated both to plasma viral levels and a subset of infected/activated monocytes (CD14+CD16+), which are known to traffic the virus into the brain. Treatment with either cART or minocycline decreased brain viral levels and partially reversed alterations in in vivo and immunohistochemical markers for neuronal injury. These findings suggest there is significant turnover of replicating virus within the brain and the severity of neuronal injury is directly related to the brain viral load

Acute Schistosoma mansoni Infection Increases Susceptibility to Systemic SHIV Clade C Infection in Rhesus Macaques after Mucosal Virus Exposure

To test the hypothesis that infection with helmiths may increase host susceptibility to infection with HIV-1, we quantified the amount of a clade C simian-human immunodeficiency virus needed to infect rhesus macaques that had acute Schistosoma mansoni infections. Compared to control animals exposed to virus alone, monkeys with schistosomiasis required exposure to 17-fold lower levels of virus to become infected. The schistosome-infected monkeys also had significantly higher levels of initial virus replication and loss of a certain subset of memory T cells, both predictors of a more rapid progression to immune dysfunction. These results suggest that worm infections may increase the risk of becoming infected with HIV-1 among individuals with viral exposures. Furthermore, they support the idea that control programs for schistosomiasis and perhaps other parasitic worm infections may also be useful in helping to reduce the spread of HIV/AIDS in developing countries where helminths are endemic

Minocycline Inhibition of Monocyte Activation Correlates with Neuronal Protection in SIV NeuroAIDS

Background: Minocycline is a tetracycline antibiotic that has been proposed as a potential conjunctive therapy for HIV-1 associated cognitive disorders. Precise mechanism(s) of minocycline’s functions are not well defined. Methods: Fourteen rhesus macaques were SIV infected and neuronal metabolites measured by proton magnetic resonance spectroscopy (1H MRS). Seven received minocycline (4 mg/kg) daily starting at day 28 post-infection (pi). Monocyte expansion and activation were assessed by flow cytometry, cell traffic to lymph nodes, CD16 regulation, viral replication, and cytokine production were studied. Results: Minocycline treatment decreased plasma virus and pro-inflammatory CD14+CD16+ and CD14loCD16+ monocytes, and reduced their expression of CD11b, CD163, CD64, CCR2 and HLA-DR. There was reduced recruitment of monocyte/ macrophages and productively infected cells in axillary lymph nodes. There was an inverse correlation between brain NAA/ Cr (neuronal injury) and circulating CD14+CD16+ and CD14loCD16+ monocytes. Minocycline treatment in vitro reduced SIV replication CD16 expression on activated CD14+CD16+ monocytes, and IL-6 production by monocytes following LPS stimulation. Conclusion: Neuroprotective effects of minocycline are due in part to reduction of activated monocytes, monocyte traffic. Mechanisms for these effects include CD16 regulation, reduced viral replication, and inhibited immune activation. Citation: Campbell JH, Burdo TH, Autissier P, Bombardier JP, Westmoreland SV, et al. (2011) Minocycline Inhibition of Monocyte Activation Correlate

Microbial Translocation Is Associated with Increased Monocyte Activation and Dementia in AIDS Patients

Elevated plasma lipopolysaccharide (LPS), an indicator of microbial translocation from the gut, is a likely cause of systemic immune activation in chronic HIV infection. LPS induces monocyte activation and trafficking into brain, which are key mechanisms in the pathogenesis of HIV-associated dementia (HAD). To determine whether high LPS levels are associated with increased monocyte activation and HAD, we obtained peripheral blood samples from AIDS patients and examined plasma LPS by Limulus amebocyte lysate (LAL) assay, peripheral blood monocytes by FACS, and soluble markers of monocyte activation by ELISA. Purified monocytes were isolated by FACS sorting, and HIV DNA and RNA levels were quantified by real time PCR. Circulating monocytes expressed high levels of the activation markers CD69 and HLA-DR, and harbored low levels of HIV compared to CD4+ T-cells. High plasma LPS levels were associated with increased plasma sCD14 and LPS-binding protein (LBP) levels, and low endotoxin core antibody levels. LPS levels were higher in HAD patients compared to control groups, and were associated with HAD independently of plasma viral load and CD4 counts. LPS levels were higher in AIDS patients using intravenous heroin and/or ethanol, or with Hepatitis C virus (HCV) co-infection, compared to control groups. These results suggest a role for elevated LPS levels in driving monocyte activation in AIDS, thereby contributing to the pathogenesis of HAD, and provide evidence that cofactors linked to substance abuse and HCV co-infection influence these processes

Proton Magnetic Resonance Spectroscopy Reveals Neuroprotection by Oral Minocycline in a Nonhuman Primate Model of Accelerated NeuroAIDS

Background: Despite the advent of highly active anti-retroviral therapy (HAART), HIV-associated neurocognitive disorders continue to be a significant problem. In efforts to understand and alleviate neurocognitive deficits associated with HIV, we used an accelerated simian immunodeficiency virus (SIV) macaque model of NeuroAIDS to test whether minocycline is neuroprotective against lentiviral-induced neuronal injury. Methodology/Principal Findings: Eleven rhesus macaques were infected with SIV, depleted of CD8+ lymphocytes, and studied until eight weeks post inoculation (wpi). Seven animals received daily minocycline orally beginning at 4 wpi. Neuronal integrity was monitored in vivo by proton magnetic resonance spectroscopy and post-mortem by immunohistochemistry for synaptophysin (SYN), microtubule-associated protein 2 (MAP2), and neuronal counts. Astrogliosis and microglial activation were quantified by measuring glial fibrillary acidic protein (GFAP) and ionized calcium binding adaptor molecule 1 (IBA-1), respectively. SIV infection followed by CD8+ cell depletion induced a progressive decline in neuronal integrity evidenced by declining N-acetylaspartate/creatine (NAA/Cr), which was arrested with minocycline treatment. The recovery of this ratio was due to increases in NAA, indicating neuronal recovery, and decreases in Cr, likely reflecting downregulation of glial cell activation. SYN, MAP2, and neuronal counts were found to be higher in minocycline-treated animals compared to untreated animals while GFAP and IBA-1 expression were decreased compared to controls. CSF and plasma viral loads were lower in MN-treated animals. Conclusions/Significance: In conclusion, oral minocycline alleviates neuronal damage induced by the AIDS virus

Phénotypage des cellules immunitaires par cytométrie en flux multiparamétrique : un outil indispensable dans l’immunopathologie du Sida

Monitoring changes in immune cell populations such as lymphocytes, monocytes and dendritic cells (DC) during infectious diseases like human immunodeficiency virus (HIV) or its counterpart in rhesus monkeys (SIV) is crucial. Thanks to recent technological advances in flow cytometry, it is now possible to measure and analyze simultaneously up to 14 individual parameters at the single cell level.The goal of this work is to develop 2 multicolor flow cytometry panels comprising of 12 antibodies, allowing measuring simultaneously the main immune cells population, respectively in humans and rhesus monkeys. After 2 years of development and optimization, we can now measure precisely all the main actors of the immune system, that is CD4+ and CD8+ T lymphocytes, B lymphocytes, NK and NKT cells, the 3 monocyte subsets, and all the dendritic cell subsets known today, by using a multicolor flow cytometry approach. This assay is done on whole blood, it is rapid to do, it does not involve a cell isolation technique, and it requires only a minimum amount of blood. Moreover, the analysis of each population is much more precise because of a minimum contamination between different cell populations. The advantage of this work is to study interactions between different cell populations of immune cells during HIV infection in humans, or SIV infection in monkeys, or potentially other diseases, and in particular to better understand the important role that dendritic cells might play in disease progression.Le suivi des changements dans les populations de cellules immunitaires tels que les lymphocytes, monocytes et cellules dendritiques (DC) au cours de maladies infectieuses comme le virus de l'immunodéficience humaine (VIH) chez l’homme ou son équivalent chez le singe (VIS) est crucial. Grâce aux récentes avancées technologiques en cytométrie en flux, il est maintenant possible de mesurer et d’analyser simultanément jusqu'à 14 paramètres individuels à l’échelon cellulaire. L'objectif de ce travail consiste en la mise au point de 2 panels multicouleurs de 12 anticorps permettant d'analyser simultanément les principales populations de cellules immunitaires, respectivement chez l’humain et le macaque rhésus. Au terme de ce travail, il est maintenant possible de mesurer précisément tous les principaux acteurs du système immunitaire, à savoir les lymphocytes T CD4+ et T CD8+, les lymphocytes B, les cellules NK et NKT, les sous-populations de monocytes, et toutes les sous-populations de cellules dendritiques connues à ce jour, en utilisant une approche multiparamétrique de cytométrie en flux. Ce protocole d’analyse est réalisé sur du sang total, il est rapide, il n’implique pas de technique d’isolation cellulaire, et requiert une quantité minimum de sang. De plus, l’analyse de chaque population cellulaire est plus précise grâce à une contamination minimum entre les populations séparées. L’intérêt de ce travail est d’étudier les interactions entre les différentes populations de cellules immunitaires durant l’infection par VIH chez l’homme, ou VIS chez le singe ou potentiellement d‘autres maladies, et en particulier de mieux comprendre le rôle important que les cellules dendritiques jouent dans la progression de ces maladies

Immunophenotyping of cell subsets by multicolor flow cytometry : an invaluable tool in the Immunopathology of AIDS

Le suivi des changements dans les populations de cellules immunitaires tels que les lymphocytes, monocytes et cellules dendritiques (DC) au cours de maladies infectieuses comme le virus de l'immunodéficience humaine (VIH) chez l’homme ou son équivalent chez le singe (VIS) est crucial. Grâce aux récentes avancées technologiques en cytométrie en flux, il est maintenant possible de mesurer et d’analyser simultanément jusqu'à 14 paramètres individuels à l’échelon cellulaire. L'objectif de ce travail consiste en la mise au point de 2 panels multicouleurs de 12 anticorps permettant d'analyser simultanément les principales populations de cellules immunitaires, respectivement chez l’humain et le macaque rhésus. Au terme de ce travail, il est maintenant possible de mesurer précisément tous les principaux acteurs du système immunitaire, à savoir les lymphocytes T CD4+ et T CD8+, les lymphocytes B, les cellules NK et NKT, les sous-populations de monocytes, et toutes les sous-populations de cellules dendritiques connues à ce jour, en utilisant une approche multiparamétrique de cytométrie en flux. Ce protocole d’analyse est réalisé sur du sang total, il est rapide, il n’implique pas de technique d’isolation cellulaire, et requiert une quantité minimum de sang. De plus, l’analyse de chaque population cellulaire est plus précise grâce à une contamination minimum entre les populations séparées. L’intérêt de ce travail est d’étudier les interactions entre les différentes populations de cellules immunitaires durant l’infection par VIH chez l’homme, ou VIS chez le singe ou potentiellement d‘autres maladies, et en particulier de mieux comprendre le rôle important que les cellules dendritiques jouent dans la progression de ces maladies.Monitoring changes in immune cell populations such as lymphocytes, monocytes and dendritic cells (DC) during infectious diseases like human immunodeficiency virus (HIV) or its counterpart in rhesus monkeys (SIV) is crucial. Thanks to recent technological advances in flow cytometry, it is now possible to measure and analyze simultaneously up to 14 individual parameters at the single cell level.The goal of this work is to develop 2 multicolor flow cytometry panels comprising of 12 antibodies, allowing measuring simultaneously the main immune cells population, respectively in humans and rhesus monkeys. After 2 years of development and optimization, we can now measure precisely all the main actors of the immune system, that is CD4+ and CD8+ T lymphocytes, B lymphocytes, NK and NKT cells, the 3 monocyte subsets, and all the dendritic cell subsets known today, by using a multicolor flow cytometry approach. This assay is done on whole blood, it is rapid to do, it does not involve a cell isolation technique, and it requires only a minimum amount of blood. Moreover, the analysis of each population is much more precise because of a minimum contamination between different cell populations. The advantage of this work is to study interactions between different cell populations of immune cells during HIV infection in humans, or SIV infection in monkeys, or potentially other diseases, and in particular to better understand the important role that dendritic cells might play in disease progression

Phénotypage des cellules immunitaires par cytométrie en flux multiparamétrique : un outil indispensable dans l’immunopathologie du Sida

Monitoring changes in immune cell populations such as lymphocytes, monocytes and dendritic cells (DC) during infectious diseases like human immunodeficiency virus (HIV) or its counterpart in rhesus monkeys (SIV) is crucial. Thanks to recent technological advances in flow cytometry, it is now possible to measure and analyze simultaneously up to 14 individual parameters at the single cell level.The goal of this work is to develop 2 multicolor flow cytometry panels comprising of 12 antibodies, allowing measuring simultaneously the main immune cells population, respectively in humans and rhesus monkeys. After 2 years of development and optimization, we can now measure precisely all the main actors of the immune system, that is CD4+ and CD8+ T lymphocytes, B lymphocytes, NK and NKT cells, the 3 monocyte subsets, and all the dendritic cell subsets known today, by using a multicolor flow cytometry approach. This assay is done on whole blood, it is rapid to do, it does not involve a cell isolation technique, and it requires only a minimum amount of blood. Moreover, the analysis of each population is much more precise because of a minimum contamination between different cell populations. The advantage of this work is to study interactions between different cell populations of immune cells during HIV infection in humans, or SIV infection in monkeys, or potentially other diseases, and in particular to better understand the important role that dendritic cells might play in disease progression.Le suivi des changements dans les populations de cellules immunitaires tels que les lymphocytes, monocytes et cellules dendritiques (DC) au cours de maladies infectieuses comme le virus de l'immunodéficience humaine (VIH) chez l’homme ou son équivalent chez le singe (VIS) est crucial. Grâce aux récentes avancées technologiques en cytométrie en flux, il est maintenant possible de mesurer et d’analyser simultanément jusqu'à 14 paramètres individuels à l’échelon cellulaire. L'objectif de ce travail consiste en la mise au point de 2 panels multicouleurs de 12 anticorps permettant d'analyser simultanément les principales populations de cellules immunitaires, respectivement chez l’humain et le macaque rhésus. Au terme de ce travail, il est maintenant possible de mesurer précisément tous les principaux acteurs du système immunitaire, à savoir les lymphocytes T CD4+ et T CD8+, les lymphocytes B, les cellules NK et NKT, les sous-populations de monocytes, et toutes les sous-populations de cellules dendritiques connues à ce jour, en utilisant une approche multiparamétrique de cytométrie en flux. Ce protocole d’analyse est réalisé sur du sang total, il est rapide, il n’implique pas de technique d’isolation cellulaire, et requiert une quantité minimum de sang. De plus, l’analyse de chaque population cellulaire est plus précise grâce à une contamination minimum entre les populations séparées. L’intérêt de ce travail est d’étudier les interactions entre les différentes populations de cellules immunitaires durant l’infection par VIH chez l’homme, ou VIS chez le singe ou potentiellement d‘autres maladies, et en particulier de mieux comprendre le rôle important que les cellules dendritiques jouent dans la progression de ces maladies

Phénotypage des cellules immunitaires par cytométrie en flux multiparamétrique (un outil indispensable dans l'immunopathologie du Sida)

Le suivi des changements dans les populations de cellules immunitaires tels que les lymphocytes, monocytes et cellules dendritiques (DC) au cours de maladies infectieuses comme le virus de l'immunodéficience humaine (VIH) chez l homme ou son équivalent chez le singe (VIS) est crucial. Grâce aux récentes avancées technologiques en cytométrie en flux, il est maintenant possible de mesurer et d analyser simultanément jusqu'à 14 paramètres individuels à l échelon cellulaire. L'objectif de ce travail consiste en la mise au point de 2 panels multicouleurs de 12 anticorps permettant d'analyser simultanément les principales populations de cellules immunitaires, respectivement chez l humain et le macaque rhésus. Au terme de ce travail, il est maintenant possible de mesurer précisément tous les principaux acteurs du système immunitaire, à savoir les lymphocytes T CD4+ et T CD8+, les lymphocytes B, les cellules NK et NKT, les sous-populations de monocytes, et toutes les sous-populations de cellules dendritiques connues à ce jour, en utilisant une approche multiparamétrique de cytométrie en flux. Ce protocole d analyse est réalisé sur du sang total, il est rapide, il n implique pas de technique d isolation cellulaire, et requiert une quantité minimum de sang. De plus, l analyse de chaque population cellulaire est plus précise grâce à une contamination minimum entre les populations séparées. L intérêt de ce travail est d étudier les interactions entre les différentes populations de cellules immunitaires durant l infection par VIH chez l homme, ou VIS chez le singe ou potentiellement d autres maladies, et en particulier de mieux comprendre le rôle important que les cellules dendritiques jouent dans la progression de ces maladies.Monitoring changes in immune cell populations such as lymphocytes, monocytes and dendritic cells (DC) during infectious diseases like human immunodeficiency virus (HIV) or its counterpart in rhesus monkeys (SIV) is crucial. Thanks to recent technological advances in flow cytometry, it is now possible to measure and analyze simultaneously up to 14 individual parameters at the single cell level.The goal of this work is to develop 2 multicolor flow cytometry panels comprising of 12 antibodies, allowing measuring simultaneously the main immune cells population, respectively in humans and rhesus monkeys. After 2 years of development and optimization, we can now measure precisely all the main actors of the immune system, that is CD4+ and CD8+ T lymphocytes, B lymphocytes, NK and NKT cells, the 3 monocyte subsets, and all the dendritic cell subsets known today, by using a multicolor flow cytometry approach. This assay is done on whole blood, it is rapid to do, it does not involve a cell isolation technique, and it requires only a minimum amount of blood. Moreover, the analysis of each population is much more precise because of a minimum contamination between different cell populations. The advantage of this work is to study interactions between different cell populations of immune cells during HIV infection in humans, or SIV infection in monkeys, or potentially other diseases, and in particular to better understand the important role that dendritic cells might play in disease progression.PARIS-CNAM (751032301) / SudocSudocFranceF