Abstract
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 asked if microglia respond to HIV infection similarly by modifying the expression, secretion, and neurotoxic potential of cathepsin B and determined the in vivo relevance of these findings. HIV-1ADA-infected human primary microglia and CHME-5 microglia cell line were assessed for expression and activity of cathepsin B, its inhibitors, cystatins B and C, and the 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 HIV encephalitis (HIVE) patients. 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 day 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. 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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Acknowledgments
We thank Dr. Randall L. Davis from the Oklahoma State University Center for Health Sciences for kindly providing the CHME-5 microglia cell line. We thank Dr. Howard Gendelman from the University of Nebraska Medical Center for providing HIV-ADA isolates. We would like to thank Rafael Romeu for his help with cell cultures and Dr. Yamil Gerena for the advice in flow cytometry. We would like to acknowledge Eduard Guerrero, Carol Torres from the RCMI Emergent Disease Core Laboratories, and Dr. Juliana Perez Laspiur and Yolanda Rodriguez from the Proteomics Core Laboratory for all their excellent technical support. Part of this work was previously presented as a poster at the ISNV. May 28-June 2, 2012, NY. Part of this work was also presented at the Annual Biomedical Research Conference for Minority Students (ABRCMS) meeting, November 13–16, 2013, Nashville, TN, USA
Funding
This work was supported in part by grants from the National Institutes of Health R01MH083516 (to LMM), U54NS4301 (to LMM), R25GM061838 (to FZ, ER, YC), GM08224, and NIMHHD8G12-MD007600 (Emergent Diseases Core and Translational Proteomics Center), and SC1GM11369-01. We acknowledge the facilities from grants ISI0 RR-13705-01 and DBI-0923132 to establish and upgrade the Confocal Microscopy Facility at the University of Puerto Rico (CIF-UPR). UPR Vice President (M.P.). UPR Medical Sciences Campus Chancellor, and the Associate Deanship of Biomedical Sciences provided additional funding to complete this study. This publication was made possible from NIH funding through the NIMH and NINDS Institutes by the following grants: Manhattan HIV Brain Bank U01MH083501, R24MH59724; Texas NeuroAIDS Research Center U01MH083507, R24 NS45491; National Neurological AIDS Bank 5U01MH083500, NS 38841; California NeuroAIDS Tissue Network U01MH083506, R24MH59745; and Statistics and Data Coordinating Center U01MH083545, N01MH32002. Its contents are solely the responsibility of the authors and do not necessarily represent the official view of the NNTC or NIH.
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Zenón, F., Cantres-Rosario, Y., Adiga, R. et al. HIV-infected microglia mediate cathepsin B-induced neurotoxicity. J. Neurovirol. 21, 544–558 (2015). https://doi.org/10.1007/s13365-015-0358-7
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DOI: https://doi.org/10.1007/s13365-015-0358-7