title: Analysis of cytokine production in a newly developed canine tracheal epithelial cell line infected with H3N2 canine influenza virus.
abstract: The Madin-Darby canine kidney (MDCK) cell line is typically used to analyze pathological features after canine influenza virus (CIV) infection. However, MDCK cells are not the ideal cell type, because they are kidney epithelial cells. Therefore, we generated an immortalized canine tracheal epithelial cell line, KU-CBE, to more reliably study immune responses to CIV infection in the respiratory tract. KU-CBE cells expressed the influenza virus receptor, α-2,3-sialic acid (SA), but not α-2,6-SA. KU-CBE and MDCK cells infected with H3N2 CIV demonstrated comparable virus growth kinetics. Gene expression levels of interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-8, IL-10, tumor necrosis factor (TNF)-α, and interferon (IFN)-β were estimated in both KU-CBE and MDCK cells infected with CIV by real-time reverse transcription polymerase chain reaction (qRT-PCR). Of these cytokines, IL-4, IL-10, TNF-α, and IFN-β mRNAs were detected in both cell lines. Gene expression of IL-4, IL-10, and TNF-α was not significantly different in the two cell lines. However, MDCK cells exhibited a significantly higher level of IFN-β mRNA than KU-CBE cells at 18 h post infection. Additionally, the protein concentrations of these four cytokines were determined by enzyme-linked immunosorbent assay (ELISA) using cell culture supernatants obtained from the two CIV-infected cell lines. MDCK cells produced significantly higher amounts of IL-4 and IFN-β than KU-CBE cells. However, KU-CBE cells produced a significantly higher amount of TNF-α than MDCK cells. These data indicated that the newly developed canine tracheal epithelial cells exhibited different cytokine production patterns compared to MDCK cells when infected with CIV. Inflammation of the respiratory tract of dogs induced by CIV infection may be attributed to the elevated expression level of TNF-α in canine tracheal epithelial cells.
authors: Department of Infectious Diseases, College of Veterinary Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 143-701, Korea.
authors: Woo-Jung
authors: WJ
authors: Park
authors: Byung-Joo
authors: BJ
authors: Park
authors: Young-Jo
authors: YJ
authors: Song
authors: Dong-Hun
authors: DH
authors: Lee
authors: Seong-Su
authors: SS
authors: Yuk
authors: Joong-Bok
authors: JB
authors: Lee
authors: Seung-Yong
authors: SY
authors: Park
authors: Chang-Seon
authors: CS
authors: Song
authors: Sang-Won
authors: SW
authors: Lee
authors: In-Soo
authors: IS
authors: Choi
mesh_terms: D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis / Q000502:physiology*; D004283:Dog Diseases / Q000276:immunology / Q000821:virology*; D004285:Dogs; D053122:Influenza A Virus, H3N2 Subtype; D016899:Interferon-beta / Q000096:biosynthesis / Q000502:physiology; D016753:Interleukin-10 / Q000096:biosynthesis / Q000502:physiology; D053583:Interleukin-1beta / Q000096:biosynthesis / Q000502:physiology; D007376:Interleukin-2 / Q000096:biosynthesis / Q000502:physiology; D015847:Interleukin-4 / Q000096:biosynthesis / Q000502:physiology; D015850:Interleukin-6 / Q000096:biosynthesis / Q000502:physiology; D016209:Interleukin-8 / Q000096:biosynthesis / Q000502:physiology; D061985:Madin Darby Canine Kidney Cells / Q000276:immunology / Q000821:virology; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D020545:Respiratory Mucosa / Q000166:cytology* / Q000503:physiopathology / Q000821:virology; D014132:Trachea / Q000166:cytology* / Q000503:physiopathology / Q000821:virology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000502:physiology
chemical_list: D016207:Cytokines; D053583:Interleukin-1beta; D007376:Interleukin-2; D015850:Interleukin-6; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D015847:Interleukin-4; D016899:Interferon-beta