title: HIV-1 gp120 as well as alcohol affect blood-brain barrier permeability and stress fiber formation: involvement of reactive oxygen species.
abstract: BACKGROUND
  HIV-1 infection commonly leads to serious HIV-1-associated neurological disorders, such as HIV-1-associated encephalopathy and dementia. In addition, alcohol is commonly used and/or abused among AIDS patients, but it is unclear whether alcohol affects the disease progression and if it affects it, how this occurs. We hypothesized that alcohol could affect the blood-brain barrier (BBB) integrity and thus could affect the onset and/or progression of HIV-associated neurological disorders.
  
  
  METHODS
  Human brain microvascular endothelial cells (HBMEC) in a BBB model system were pretreated with alcohol (17 and 68 mM) and subsequently coexposed with HIV-1 gp120. Expression of chemokine receptors CCR3, CCR5, and CXCR4 was assessed by enzyme-linked immunosorbent assay and real-time polymerase chain reaction. Changes in the permeability of the HBMEC monolayer were assessed using paracellular markers [(3)H]inulin or propidium iodide. Actin rearrangements in HBMEC were visualized by fluorescence microscopy and viability assessed using Live/Dead stain.
  
  
  RESULTS
  Both gp120 and alcohol increased the permeability of the BBB model by up to 141%, without affecting HBMEC viability. Cotreatment with alcohol and gp120 did not result in a significant synergistic effect. Gp120 permeability involved chemokine receptor CCR5. Alcohol did not affect chemokine receptor expression on brain endothelial cells. Both gp120 and alcohol reorganized the cytoskeleton and induced stress fiber formation. Inhibition of reactive oxygen species (ROS) formation through NADPH blocked the effects of both gp120 and alcohol on permeability and stress fiber formation.
  
  
  CONCLUSIONS
  These results indicate that both HIV-1 gp120 and alcohol induce stress fibers, causing increased permeability of the human BBB endothelium. Alcohol (68 mM)-mediated permeability increase was linked to ROS formation. The alcohol-mediated physiological changes in the HBMEC monolayers may increase diffusion of plasma components and viral penetration across the BBB. This suggests that alcohol, especially at levels attained in heavy drinkers, can potentially contribute in a negative fashion to HIV-1 neuropathogenesis.
authors.affiliation: Department of Pediatrics, Division Infectious Diseases, Johns Hopkins University School of Medicine, 720 Rutland Avenue, Baltimore, MD 21205, USA.
authors.forename: Carlum
authors.initials: C
authors.lastname: Shiu
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Barbier
authors.forename: Francescopaolo
authors.initials: F
authors.lastname: Di Cello
authors.forename: Hee Jung
authors.initials: HJ
authors.lastname: Choi
authors.forename: Monique
authors.initials: M
authors.lastname: Stins
mesh_terms: D000465:Algorithms; D001812:Blood-Brain Barrier / Q000187:drug effects* / Q000473:pathology*; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D000431:Ethanol / Q000494:pharmacology*; D015699:HIV Envelope Protein gp120 / Q000494:pharmacology*; D006801:Humans; D007202:Indicators and Reagents; D017382:Reactive Oxygen Species / Q000378:metabolism*; D019707:Receptors, Chemokine / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D022502:Stress Fibers / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*
chemical_list: D002492:Central Nervous System Depressants; D015699:HIV Envelope Protein gp120; D007202:Indicators and Reagents; D017382:Reactive Oxygen Species; D019707:Receptors, Chemokine; D000431:Ethanol