title: Monocytes-derived macrophages mediated stable expression of human brain-derived neurotrophic factor, a novel therapeutic strategy for neuroAIDS.
abstract: HIV-1 associated dementia remains a significant public health burden. Clinical and experimental research has shown that reduced levels of brain-derived neurotrophic factor (BDNF) may be a risk factor for neurological complications associated with HIV-1 infection. We are actively testing genetically modified macrophages for their possible use as the cell-based gene delivery vehicle for the central nervous system (CNS). It can be an advantage to use the natural homing/migratory properties of monocyte-derived macrophages to deliver potentially neuroprotective BDNF into the CNS, as a non-invasive manner. Lentiviral-mediated gene transfer of human (h)BDNF plasmid was constructed and characterized. Defective lentiviral stocks were generated by transient transfection of 293T cells with lentiviral transfer plasmid together with packaging and envelope plasmids. High titer lentiviral vector stocks were harvested and used to transduce human neuronal cell lines, primary cultures of human peripheral mononocyte-derived macrophages (hMDM) and murine myeloid monocyte-derived macrophages (mMDM). These transduced cells were tested for hBDNF expression, stability, and neuroprotective activity. The GenomeLab GeXP Genetic Analysis System was used to evaluate transduced cells for any adverse effects by assessing gene profiles of 24 reference genes. High titer vectors were prepared for efficient transduction of neuronal cell lines, hMDM, and mMDM. Stable secretion of high levels of hBDNF was detected in supernatants of transduced cells using western blot and ELISA. The conditioned media containing hBDNF were shown to be protective to neuronal and monocytic cell lines from TNF-α and HIV-1 Tat mediated cytotoxicity. Lentiviral vector-mediated gene transduction of hMDM and mMDM resulted in high-level, stable expression of the neuroprotective factorBDNF in vitro. These findings form the basis for future research on the potential use of BDNF as a novel therapy for neuroAIDS.
authors: MOE Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Institute of TCM & Natural Products, School of Pharmaceutical Sciences, Wuhan University, Wuhan, People's Republic of China ; Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, Hawaii, United States of America.
authors: Jing
authors: J
authors: Tong
authors: University of Nebraska Medical Center, Pharmacology and Experimental Neuroscience, Nebraska Medical Center, Omaha, Nebraska, United States of America.
authors: Shilpa
authors: S
authors: Buch
authors: University of Nebraska Medical Center, Pharmacology and Experimental Neuroscience, Nebraska Medical Center, Omaha, Nebraska, United States of America.
authors: Honghong
authors: H
authors: Yao
authors: Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, Hawaii, United States of America.
authors: Chengxiang
authors: C
authors: Wu
authors: Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, Hawaii, United States of America.
authors: Hsin-I
authors: HI
authors: Tong
authors: MOE Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Institute of TCM & Natural Products, School of Pharmaceutical Sciences, Wuhan University, Wuhan, People's Republic of China.
authors: Youwei
authors: Y
authors: Wang
authors: Department of Public Health Sciences, University of Hawaii at Manoa, Honolulu, Hawaii, United States of America.
authors: Yuanan
authors: Y
authors: Lu
mesh_terms: D015526:AIDS Dementia Complex / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000096:biosynthesis* / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006801:Humans; D016086:Lentivirus; D008264:Macrophages / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D009000:Monocytes / Q000378:metabolism* / Q000473:pathology; D014161:Transduction, Genetic
chemical_list: D019208:Brain-Derived Neurotrophic Factor; C579455:BDNF protein, human