title: Immunohistochemical and in situ hybridization studies of influenza A virus infection in human lungs.
abstract: Influenza viruses are responsible for acute febrile respiratory disease. When deaths occur, definitive diagnosis requires viral isolation because no characteristic viral inclusions are seen. We examined the distribution of influenza A virus in tissues from 8 patients with fatal infection using 2 immunohistochemical assays (monoclonal antibodies to nucleoprotein [NP] and hemagglutinin [HA]) and 2 in situ hybridization (ISH) assays (digoxigenin-labeled probes that hybridized to HA and NP genes). Five patients had prominent bronchitis; by immunohistochemical assay, influenza A staining was present focally in the epithelium of larger bronchi (intact and detached necrotic cells) and in rare interstitial cells. The anti-NP antibody stained primarily cell nuclei, and the anti-HA antibody stained mainly the cytoplasm. In 4 of these cases, nucleic acids (ISH) were identified in the same areas. Three patients had lymphohistiocytic alveolitis and showed no immunohistochemical or ISH staining. Both techniques were useful for detection of influenza virus antigens and nucleic acids in formalin-fixed paraffin-embedded tissues and can enable further understanding of fatal influenza A virus infections in humans.
authors.affiliation: Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.
authors.forename: J
authors.initials: J
authors.lastname: Guarner
authors.forename: W J
authors.initials: WJ
authors.lastname: Shieh
authors.forename: J
authors.initials: J
authors.lastname: Dawson
authors.forename: K
authors.initials: K
authors.lastname: Subbarao
authors.forename: M
authors.initials: M
authors.lastname: Shaw
authors.forename: T
authors.initials: T
authors.lastname: Ferebee
authors.forename: T
authors.initials: T
authors.lastname: Morken
authors.forename: K B
authors.initials: KB
authors.lastname: Nolte
authors.forename: A
authors.initials: A
authors.lastname: Freifeld
authors.forename: N
authors.initials: N
authors.lastname: Cox
authors.forename: S R
authors.initials: SR
authors.lastname: Zaki
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001991:Bronchitis / Q000473:pathology / Q000821:virology; D002648:Child; D005260:Female; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D017403:In Situ Hybridization; D009980:Influenza A virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D007251:Influenza, Human / Q000473:pathology / Q000821:virology*; D008168:Lung / Q000473:pathology / Q000821:virology*; D017563:Lung Diseases, Interstitial / Q000473:pathology / Q000821:virology; D008297:Male; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000235:genetics / Q000276:immunology; D016612:Paraffin Embedding; D012367:RNA, Viral / Q000032:analysis; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins