title: Cytokine Expression at Different Stages of Influenza A(H1N1)pdm09 Virus Infection in the Porcine Lung, Using Laser Capture Microdissection.
abstract: Pandemic influenza A(H1N1)pdm09 virus has retained its ability to infect swine whilst developing the ability to transmit effectively between humans, thus making the pig a valuable model for studying disease pathogenesis in both species. Lung lesions in pigs caused by infection with influenza A viruses vary in both their severity and distribution with individual lung lobes exhibiting lesions at different stages of infection pathogenic development and disease resolution. Consequently, investigating interactions between the virus and host and their implications for disease pathogenesis can be complicated. Studies were undertaken to investigate the discrete expression of pro- and anti-inflammatory mediators during lung lesion formation in pigs during infection with influenza A(H1N1)pdm09 (A/Hamburg/05/09) virus. Laser capture microdissection was used to identify and select lung lobules containing lesions at different stages of development. Dissected samples were analysed using quantitative RT-PCR to assess pro- and anti-inflammatory cytokine mRNA transcripts. Differential expression of the immune mediators IL-8, IL-10 and IFN-γ was observed depending upon the lesion stage assessed. Upregulation of IFN-γ, IL-8 and IL-10 mRNA was observed in stage 2 lesions, whereas decreased mRNA expression was observed in stage 3 lesions, with IL-8 actively downregulated when compared with controls in both stage 3 and stage 4 lesions. This study highlighted the value of using laser capture microdissection to isolate specific tissue regions and investigate subtle differences in cytokine mRNA expression during lesion development in pigs infected with influenza A(H1N1)pdm09.
authors.affiliation: Pathology Department, Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Addlestone, Surrey, UK.
authors.forename: D J
authors.initials: DJ
authors.lastname: Hicks
authors.affiliation: Virology Department, Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Addlestone, Surrey, UK.
authors.forename: M
authors.initials: M
authors.lastname: Kelly
authors.affiliation: Virology Department, Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Addlestone, Surrey, UK.
authors.forename: S M
authors.initials: SM
authors.lastname: Brookes
authors.affiliation: Virology Department, Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Addlestone, Surrey, UK.
authors.forename: B Z
authors.initials: BZ
authors.lastname: Londt
authors.affiliation: Centre for Epidemiology and Risk Analysis, Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Addlestone, Surrey, UK.
authors.forename: A
authors.initials: A
authors.lastname: Ortiz Pelaez
authors.affiliation: Pathology Department, Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Addlestone, Surrey, UK.
authors.forename: A
authors.initials: A
authors.lastname: Orlowska
authors.affiliation: Virology Department, Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Addlestone, Surrey, UK.
authors.forename: I H
authors.initials: IH
authors.lastname: Brown
authors.affiliation: Pathology Department, Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Addlestone, Surrey, UK.
authors.forename: Y I
authors.initials: YI
authors.lastname: Spencer
authors.affiliation: Pathology Department, Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Addlestone, Surrey, UK.
authors.forename: A
authors.initials: A
authors.lastname: Núñez
mesh_terms: D000818:Animals; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology; D009980:Influenza A virus / Q000235:genetics; D016753:Interleukin-10; D061346:Laser Capture Microdissection; D008168:Lung / Q000378:metabolism* / Q000473:pathology; D009976:Orthomyxoviridae Infections / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013552:Swine; D013553:Swine Diseases / Q000378:metabolism / Q000821:virology*
chemical_list: D016207:Cytokines; C508609:IL10 protein, human; D012333:RNA, Messenger; D016753:Interleukin-10
keywords: cytokines; influenza A; laser capture microdissection; pandemic H1N1; pig