title: Microglia-derived HIV Nef+ exosome impairment of the blood-brain barrier is treatable by nanomedicine-based delivery of Nef peptides.
abstract: The negative factor (Nef) of human immunodeficiency virus (HIV) is an accessory protein that is thought to be integral to HIV-associated immune- and neuroimmune pathogenesis. Here, we show that nef-transfected microglia-released Nef+ exosome (exNef) disrupts the apical blood-brain barrier (BBB) and that only nef-transfected microglia release Nef in exosomes. nef-gfp-transduced neurons and astrocytes release exosomes but did not release exNef in the extracellular space. Apical administration of exNef derived from nef-transfected 293T cells reduced transendothelial electrical resistance (TEER) and increased permeability of the BBB. Microglia-derived exNef applied to either the apical/basal BBB significantly reduced expression of the tight junction protein, ZO-1, suggesting a mechanism of exNef-mediated neuropathogenesis. Microglia exposed to exNef release elevated levels of Toll-like receptor-induced cytokines and chemokines IL-12, IL-8, IL-6, RANTES, and IL-17A. Magnetic nanoparticle delivery of Nef peptides containing the Nef myrisolation site across an in vitro BBB ultimately reduced nef-transfected microglia release of Nef exosomes and prevented the loss of BBB integrity and permeability as measured by TEER and dextran-FITC transport studies, respectively. Overall, we show that exNef is released from nef-gfp-transfected microglia; exNef disrupts integrity and permeability, and tight junctions of the BBB, and induces microglial cytokine/chemokine secretion. These exNef-mediated effects were significantly restricted by Nef peptides. Taken together, this study provides preliminary evidence of the role of exNef in HIV neuroimmune pathogenesis and the feasibility of a nanomedicine-based therapeutics targeting exNef to treat HIV-associated neuropathogenesis.
authors.affiliation: Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA. adraymon@fiu.edu.|Institute of Neuroimmune Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA. adraymon@fiu.edu.
authors.forename: A D
authors.initials: AD
authors.lastname: Raymond
authors.affiliation: Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: P
authors.initials: P
authors.lastname: Diaz
authors.affiliation: Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: S
authors.initials: S
authors.lastname: Chevelon
authors.affiliation: Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: M
authors.initials: M
authors.lastname: Agudelo
authors.affiliation: Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.|Institute of Neuroimmune Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: A
authors.initials: A
authors.lastname: Yndart-Arias
authors.affiliation: Institute of Neuroimmune Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: H
authors.initials: H
authors.lastname: Ding
authors.affiliation: Institute of Neuroimmune Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: A
authors.initials: A
authors.lastname: Kaushik
authors.affiliation: Institute of Neuroimmune Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: R Dev
authors.initials: RD
authors.lastname: Jayant
authors.affiliation: Department of Mechanical and Materials Engineering, College of Engineering and Computing, Florida International University, 10555 W. Flagler St., Miami, FL, 33174, USA.
authors.forename: R
authors.initials: R
authors.lastname: Nikkhah-Moshaie
authors.affiliation: Institute of Neuroimmune Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: U
authors.initials: U
authors.lastname: Roy
authors.affiliation: Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: S
authors.initials: S
authors.lastname: Pilakka-Kanthikeel
authors.affiliation: Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.|Institute of Neuroimmune Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA.
authors.forename: M P
authors.initials: MP
authors.lastname: Nair
mesh_terms: D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D018946:Chemokine CCL5 / Q000235:genetics / Q000276:immunology; D004337:Drug Carriers / Q000494:pharmacology*; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D055354:Exosomes / Q000378:metabolism*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D015497:HIV-1 / Q000737:chemistry; D006801:Humans; D007378:Interleukins / Q000235:genetics / Q000276:immunology; D058185:Magnetite Nanoparticles / Q000737:chemistry; D017628:Microglia / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008954:Models, Biological; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology*; D015398:Signal Transduction; D014162:Transfection; D019076:Transgenes; D054311:nef Gene Products, Human Immunodeficiency Virus / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology
chemical_list: C483024:CCL5 protein, human; D018946:Chemokine CCL5; D004337:Drug Carriers; D007378:Interleukins; D058185:Magnetite Nanoparticles; D010455:Peptides; D054311:nef Gene Products, Human Immunodeficiency Virus; C514837:nef protein, Human immunodeficiency virus 1; D049452:Green Fluorescent Proteins
keywords: BBB impairment; Nanoparticle therapeutics; Nef + exosomes