Proteomic Analysis of HIV-Infected Macrophages

Meléndez, Loyda M.; Colón, Krystal; Rivera, Linda; Rodriguez-Franco, Eillen; Toro-Nieves, Dianedis

doi:10.1007/s11481-010-9253-4

Cited by 24 publications

(29 citation statements)

References 145 publications

(228 reference statements)

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“…We found that CHME-5 produce low levels of HIV p24 and undetectable cystatin B in supernatants which correlates with the low level of HIV-1 replication found in these cells as well as other tissue macrophages including placental macrophages (Luciano-Montalvo et al 2008; Luciano-Montalvo and Melendez 2009). In contrast high levels of secreted cystatin B at 12 dpi correlates with HIV-1 infection in MDM as reported in previous studies (Meléndez et al 2011; Rivera-Rivera et al 2012). …”

Section: Discussionsupporting

confidence: 86%

“…Proteomic analysis of HIV-infected macrophages has revealed that HIV infection can induce profound alterations in its normal cell physiology, potentially contributing to neuronal dysfunction [as reviewed by Melendez and collaborators, 2011; (Melendez et al 2011). These alterations include not only the production of neurotoxins, but also the modulation of normal cellular processes.…”

Section: Discussionmentioning

confidence: 99%

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HIV-infected microglia mediate cathepsin B-induced neurotoxicity

et al. 2015

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BACKGROUND HIV-1-infected mononuclear phagocytes release soluble factors that affect the homeostasis in tissue. HIV-1 can prompt metabolic encephalopathy with the addition of neuronal dysfunction and apoptosis. Recently, we reported that HIV-1 enhances the expression and secretion of bioactive cathepsin B in monocyte-derived macrophages, ultimately contributing to neuronal apoptosis. In this research, we request if microglia respond to HIV infection similarly by modifying the expression, secretion, neurotoxic potential of cathepsin B and the in vivo relevance of these findings. METHODS HIV-ADA infected human primary microglia and CHME-5 were assessed for expression and activity of cathepsin B, its inhibitors, cystatins B and C, and neurotoxicity associated with these changes. Human primary neurons were exposed to supernatants from HIV-infected and uninfected microglia in the presence of cathepsin B inhibitors and apoptosis was assessed by TUNEL. Microglial expression of cathepsin B was validated in brain tissue from HIVE patients. RESULTS HIV-infected microglia secreted significantly greater levels of cathepsin B, cystatin B, and cystatin C compared to uninfected cells. Increased apoptosis was observed in neurons exposed to supernatants from HIV-1 infected microglia at days 12 post-infection. The cathepsin B inhibitor CA-074 and cathepsin B antibody prevented neuronal apoptosis. Increased microglia-derived cathepsin B, cystatin B, and cystatin C and caspase-3+ neurons were detected in HIVE brains compared to controls. CONCLUSIONS Our results suggest that HIV-1-induced cathepsin B production in microglia contributes to neuronal apoptosis and may be an important factor in neuronal death associated with HIVE.

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Section: Discussionsupporting

confidence: 86%

Section: Discussionmentioning

confidence: 99%

HIV-infected microglia mediate cathepsin B-induced neurotoxicity

et al. 2015

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“…Previous proteomic studies from our laboratories identified cathepsin B and cystatin B as differentially expressed in HIV-infected macrophages [31]. To determine if HIV-1 has an effect on the expression of genes for cathepsin B and its inhibitors, cystatins B and C, we performed real time PCR of HIV-infected and uninfected MDM cultures from 8 different donors.…”

Section: Resultsmentioning

confidence: 99%

“…Proteomic analyses of HIV-infected macrophages revealed that HIV-1 infection induces profound alterations in the normal physiology of macrophages, which could contribute to neuronal dysfunction [31]. These changes include not only the production of neurotoxins, but also the dysregulation of normal cellular processes.…”

Section: Discussionmentioning

confidence: 99%

Dysregulation of Macrophage-Secreted Cathepsin B Contributes to HIV-1-Linked Neuronal Apoptosis

et al. 2012

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Chronic HIV infection leads to the development of cognitive impairments, designated as HIV-associated neurocognitive disorders (HAND). The secretion of soluble neurotoxic factors by HIV-infected macrophages plays a central role in the neuronal dysfunction and cell death associated with HAND. One potentially neurotoxic protein secreted by HIV-1 infected macrophages is cathepsin B. To explore the potential role of cathepsin B in neuronal cell death after HIV infection, we cultured HIV-1ADA infected human monocyte-derived macrophages (MDM) and assayed them for expression and activity of cathepsin B and its inhibitors, cystatins B and C. The neurotoxic activity of the secreted cathepsin B was determined by incubating cells from the neuronal cell line SK-N-SH with MDM conditioned media (MCM) from HIV-1 infected cultures. We found that HIV-1 infected MDM secreted significantly higher levels of cathepsin B than did uninfected cells. Moreover, the activity of secreted cathepsin B was significantly increased in HIV-infected MDM at the peak of viral production. Incubation of neuronal cells with supernatants from HIV-infected MDM resulted in a significant increase in the numbers of apoptotic neurons, and this increase was reversed by the addition of either the cathepsin B inhibitor CA-074 or a monoclonal antibody to cathepsin B. In situ proximity ligation assays indicated that the increased neurotoxic activity of the cathepsin B secreted by HIV-infected MDM resulted from decreased interactions between the enzyme and its inhibitors, cystatins B and C. Furthermore, preliminary in vivo studies of human post-mortem brain tissue suggested an upregulation of cathepsin B immunoreactivity in the hippocampus and basal ganglia in individuals with HAND. Our results demonstrate that HIV-1 infection upregulates cathepsin B in macrophages, increases cathepsin B activity, and reduces cystatin-cathepsin interactions, contributing to neuronal apoptosis. These findings provide new evidence for the role of cathepsin B in neuronal cell death induced by HIV-infected macrophages.

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“…Macrophage recruitment and microglial activation were observed to co-occur with HIV encephalitis and HIV-associated dementia during the pre-HAART era (Glass et al, 1995; Tyor et al, 1995) and remain distinguishing features of neuroAIDS today, despite HAART (Burdo et al, 2013; Spudich & González -Scarano, 2012). Macrophage recruitment and microglial activation are associated with the production of cytokines, chemokines, proteases, reactive oxygen and nitrosative species, in addition to several excitotoxins (Colton & Gilbert, 1987; Kaul et al, 2001; Kraft-Terry et al, 2011; Meléndez et al, 2011). In pre-HAART observations of disease progression, microglial nodules, astrocytosis, and neurodegeneration characterized the clinical histopathology accompanying HIV-associated dementia (Everall et al, 1993; Kure et al, 1991; Navia et al, 1986).…”

Section: Discussionmentioning

confidence: 99%

5α-reduced progestogens ameliorate mood-related behavioral pathology, neurotoxicity, and microgliosis associated with exposure to HIV-1 Tat

Paris

Zou

Hahn

et al. 2016

Brain, Behavior, and Immunity

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Human immunodeficiency virus (HIV) is associated with motor and mood disorders, likely influenced by reactive microgliosis and subsequent neural damage. We have recapitulated aspects of this pathology in mice that conditionally express the neurotoxic HIV-1 regulatory protein, trans-activator of transcription (Tat). Progestogens may attenuate Tat-related behavioral impairments and reduce neurotoxicity in vitro, perhaps via progesterone’s 5α-reductase-dependent metabolism to the neuroprotective steroid, allopregnanolone. To test this, ovariectomized female mice that conditionally expressed (or did not express) central HIV-1 Tat were administered vehicle or progesterone (4 mg/kg), with or without pretreatment of a 5α-reductase inhibitor (finasteride, 50 mg/kg). Tat induction significantly increased anxiety-like behavior in an open field, elevated plus maze and a marble burying task concomitant with elevated protein oxidation in striatum. Progesterone administration attenuated anxiety-like effects in the open field and elevated plus maze, but not in conjunction with finasteride pretreatment. Progesterone also attenuated Tat-promoted protein oxidation in striatum, independent of finasteride pretreatment. Concurrent experiments in vitro revealed Tat (50 nM)-mediated reductions in neuronal cell survival over 60 h, as well as increased neuronal and microglial intracellular calcium, as assessed via fura-2 AM fluorescence. Co-treatment with allopregnanolone (100 nM) attenuated neuronal death in time-lapse imaging and blocked the Tat-induced exacerbation of intracellular calcium in neurons and microglia. Lastly, neuron-glia co-cultures were labeled for Iba-1 to reveal that Tat increased microglial numbers in vitro and co-treatment with allopregnanolone attenuated this effect. Together, these data support the notion that 5α-reduced pregnane steroids exert protection over the neurotoxic effects of HIV-1 Tat.

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Proteomic Analysis of HIV-Infected Macrophages

Cited by 24 publications

References 145 publications

HIV-infected microglia mediate cathepsin B-induced neurotoxicity

HIV-infected microglia mediate cathepsin B-induced neurotoxicity

Dysregulation of Macrophage-Secreted Cathepsin B Contributes to HIV-1-Linked Neuronal Apoptosis

5α-reduced progestogens ameliorate mood-related behavioral pathology, neurotoxicity, and microgliosis associated with exposure to HIV-1 Tat

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