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Archives of Virology

, Volume 160, Issue 6, pp 1397–1405 | Cite as

Analysis of cytokine production in a newly developed canine tracheal epithelial cell line infected with H3N2 canine influenza virus

  • Woo-Jung Park
  • Byung-Joo Park
  • Young-Jo Song
  • Dong-Hun Lee
  • Seong-Su Yuk
  • Joong-Bok Lee
  • Seung-Yong Park
  • Chang-Seon Song
  • Sang-Won Lee
  • In-Soo ChoiEmail author
Original Article

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.

Keywords

Influenza Virus Sialic Acid Reverse Transcription Polymerase Chain Reaction MDCK Cell Normal Human Bronchial Epithelial 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This research was supported by the Brain Korea 21 Plus and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2013-A419-0092).

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Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Woo-Jung Park
    • 1
  • Byung-Joo Park
    • 1
  • Young-Jo Song
    • 1
  • Dong-Hun Lee
    • 1
  • Seong-Su Yuk
    • 1
  • Joong-Bok Lee
    • 1
  • Seung-Yong Park
    • 1
  • Chang-Seon Song
    • 1
  • Sang-Won Lee
    • 1
  • In-Soo Choi
    • 1
    Email author
  1. 1.Department of Infectious Diseases, College of Veterinary MedicineKonkuk UniversitySeoulKorea

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