title: Lower Respiratory Tract Infection of the Ferret by 2009 H1N1 Pandemic Influenza A Virus Triggers Biphasic, Systemic, and Local Recruitment of Neutrophils.
abstract: Infection of the lower respiratory tract by influenza A viruses results in increases in inflammation and immune cell infiltration in the lung. The dynamic relationships among the lung microenvironments, the lung, and systemic host responses during infection remain poorly understood. Here we used extensive systematic histological analysis coupled with live imaging to gain access to these relationships in ferrets infected with the 2009 H1N1 pandemic influenza A virus (H1N1pdm virus). Neutrophil levels rose in the lungs of H1N1pdm virus-infected ferrets 6 h postinfection and became concentrated at areas of the H1N1pdm virus-infected bronchiolar epithelium by 1 day postinfection (dpi). In addition, neutrophil levels were increased throughout the alveolar spaces during the first 3 dpi and returned to baseline by 6 dpi. Histochemical staining revealed that neutrophil infiltration in the lungs occurred in two waves, at 1 and 3 dpi, and gene expression within microenvironments suggested two types of neutrophils. Specifically, CCL3 levels, but not CXCL8/interleukin 8 (IL-8) levels, were higher within discrete lung microenvironments and coincided with increased infiltration of neutrophils into the lung. We used live imaging of ferrets to monitor host responses within the lung over time with [(18)F]fluorodeoxyglucose (FDG). Sites in the H1N1pdm virus-infected ferret lung with high FDG uptake had high levels of proliferative epithelium. In summary, neutrophils invaded the H1N1pdm virus-infected ferret lung globally and focally at sites of infection. Increased neutrophil levels in microenvironments did not correlate with increased FDG uptake; hence, FDG uptake may reflect prior infection and inflammation of lungs that have experienced damage, as evidenced by bronchial regeneration of tissues in the lungs at sites with high FDG levels.
  
  
  OBJECTIVE
  Severe influenza disease is characterized by an acute infection of the lower airways that may progress rapidly to organ failure and death. Well-developed animal models that mimic human disease are essential to understanding the complex relationships of the microenvironment, organ, and system in controlling virus replication, inflammation, and disease progression. Employing the ferret model of H1N1pdm virus infection, we used live imaging and comprehensive histological analyses to address specific hypotheses regarding spatial and temporal relationships that occur during the progression of infection and inflammation. We show the general invasion of neutrophils at the organ level (lung) but also a distinct pattern of localized accumulation within the microenvironment at the site of infection. Moreover, we show that these responses were biphasic within the lung. Finally, live imaging revealed an early and sustained host metabolic response at sites of infection that may reflect damage and repair of tissues in the lungs.
authors.affiliation: Department of Microbiology and Immunology, University of Louisville, Louisville, Kentucky, USA Center for Predictive Medicine for Infectious Diseases and Biodefense, University of Louisville, Louisville, Kentucky, USA.
authors.forename: Jeremy V
authors.initials: JV
authors.lastname: Camp
authors.affiliation: Center for Infectious Disease Imaging, Department of Radiology and Imaging Sciences, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Ulas
authors.initials: U
authors.lastname: Bagci
authors.affiliation: Center for Predictive Medicine for Infectious Diseases and Biodefense, University of Louisville, Louisville, Kentucky, USA.
authors.forename: Yong-Kyu
authors.initials: YK
authors.lastname: Chu
authors.affiliation: School of Dentistry, University of Louisville, Louisville, Kentucky, USA.
authors.forename: Brendan
authors.initials: B
authors.lastname: Squier
authors.affiliation: Department of Medicine, University of Louisville, Louisville, Kentucky, USA Department of Pathology, University of Louisville, Louisville, Kentucky, USA.
authors.forename: Mostafa
authors.initials: M
authors.lastname: Fraig
authors.affiliation: Department of Microbiology and Immunology, University of Louisville, Louisville, Kentucky, USA Department of Medicine, University of Louisville, Louisville, Kentucky, USA.
authors.forename: Silvia M
authors.initials: SM
authors.lastname: Uriarte
authors.affiliation: Center for Predictive Medicine for Infectious Diseases and Biodefense, University of Louisville, Louisville, Kentucky, USA Department of Radiology, University of Louisville, Louisville, Kentucky, USA.
authors.forename: Haixun
authors.initials: H
authors.lastname: Guo
authors.affiliation: Center for Infectious Disease Imaging, Department of Radiology and Imaging Sciences, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Mollura
authors.affiliation: Department of Microbiology and Immunology, University of Louisville, Louisville, Kentucky, USA Center for Predictive Medicine for Infectious Diseases and Biodefense, University of Louisville, Louisville, Kentucky, USA cjonsson@utk.edu.
authors.forename: Colleen B
authors.initials: CB
authors.lastname: Jonsson
mesh_terms: D000818:Animals; D018932:Chemokine CCL2 / Q000235:genetics / Q000276:immunology; D054405:Chemokine CCL3 / Q000235:genetics / Q000276:immunology; D005260:Female; D005289:Ferrets / Q000276:immunology / Q000821:virology; D019788:Fluorodeoxyglucose F18; D015870:Gene Expression; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology* / Q000472:pathogenicity; D016209:Interleukin-8 / Q000276:immunology; D008168:Lung / Q000166:cytology / Q000276:immunology / Q000821:virology; D020556:Neutrophil Infiltration / Q000276:immunology*; D009504:Neutrophils / Q000276:immunology*; D009976:Orthomyxoviridae Infections / Q000276:immunology* / Q000662:veterinary / Q000821:virology; D049268:Positron-Emission Tomography; D012141:Respiratory Tract Infections / Q000276:immunology* / Q000662:veterinary / Q000821:virology
chemical_list: D018932:Chemokine CCL2; D054405:Chemokine CCL3; D016209:Interleukin-8; D019788:Fluorodeoxyglucose F18