title: CCR5 limits cortical viral loads during West Nile virus infection of the central nervous system.
abstract: BACKGROUND
  Cell-mediated immunity is critical for clearance of central nervous system (CNS) infection with the encephalitic flavivirus, West Nile virus (WNV). Prior studies from our laboratory have shown that WNV-infected neurons express chemoattractants that mediate recruitment of antiviral leukocytes into the CNS. Although the chemokine receptor, CCR5, has been shown to play an important role in CNS host defense during WNV infection, regional effects of its activity within the infected brain have not been defined.
  
  
  METHODS
  We used CCR5-deficient mice and an established murine model of WNV encephalitis to determine whether CCR5 activity impacts on WNV levels within the CNS in a region-specific fashion. Statistical comparisons between groups were made with one- or two-way analysis of variance; Bonferroni's post hoc test was subsequently used to compare individual means. Survival was analyzed by the log-rank test. Analyses were conducted using Prism software (GraphPad Prism). All data were expressed as means ± SEM. Differences were considered significant if P ≤ 0.05.
  
  
  RESULTS
  As previously shown, lack of CCR5 activity led to increased symptomatic disease and mortality in mice after subcutaneous infection with WNV. Evaluation of viral burden in the footpad, draining lymph nodes, spleen, olfactory bulb, and cerebellum derived from WNV-infected wild-type, and CCR5(-/-) mice showed no differences between the genotypes. In contrast, WNV-infected, CCR5(-/-) mice exhibited significantly increased viral burden in cortical tissues, including the hippocampus, at day 8 post-infection. CNS regional studies of chemokine expression via luminex analysis revealed significantly increased expression of CCR5 ligands, CCL4 and CCL5, within the cortices of WNV-infected, CCR5(-/-) mice compared with those of similarly infected WT animals. Cortical elevations in viral loads and CCR5 ligands in WNV-infected, CCR5(-/-) mice, however, were associated with decreased numbers of infiltrating mononuclear cells and increased permeability of the blood-brain barrier.
  
  
  CONCLUSIONS
  These data indicate that regional differences in chemokine expression occur in response to WNV infection of the CNS, and that cortical neurons require CCR5 activity to limit viral burden in this brain region.
authors.affiliation: Department of Medicine, Washington University School of Medicine, St Louis, MO, 63110, USA. ddurrant@dom.wustl.edu.
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Durrant
authors.affiliation: Department of Medicine, Washington University School of Medicine, St Louis, MO, 63110, USA. bdaniels@dom.wustl.edu.
authors.forename: Brian P
authors.initials: BP
authors.lastname: Daniels
authors.affiliation: Department of Medicine, Washington University School of Medicine, St Louis, MO, 63110, USA. PasiekaT@vision.wustl.edu.
authors.forename: TracyJo
authors.initials: T
authors.lastname: Pasieka
authors.affiliation: Department of Medicine, Washington University School of Medicine, St Louis, MO, 63110, USA.
authors.forename: Denise
authors.initials: D
authors.lastname: Dorsey
authors.affiliation: Department of Medicine, Washington University School of Medicine, St Louis, MO, 63110, USA. rklein@dom.wustl.edu.|Department of Pathology & Immunology, Washington University School of Medicine, St Louis, MO, 63110, USA. rklein@dom.wustl.edu.|Department of Anatomy & Neurobiology, Washington University School of Medicine, St Louis, MO, 63110, USA. rklein@dom.wustl.edu.
authors.forename: Robyn S
authors.initials: RS
authors.lastname: Klein
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000276:immunology / Q000378:metabolism; D002490:Central Nervous System / Q000276:immunology / Q000378:metabolism; D002540:Cerebral Cortex / Q000276:immunology* / Q000378:metabolism / Q000821:virology; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism; D051379:Mice; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D019713:Receptors, CCR5 / Q000172:deficiency* / Q000276:immunology*; D019562:Viral Load / Q000502:physiology*; D014901:West Nile Fever / Q000276:immunology* / Q000378:metabolism; D014902:West Nile virus / Q000276:immunology* / Q000302:isolation & purification
chemical_list: C000592874:CCR5 protein, mouse; D019713:Receptors, CCR5