Airway inflammation induced by recombinant guinea pig tumor necrosis factor-α

Anna Marie White, Teizo Yoshimura, Anthony W. Smith, John Westwick, Malcolm L. Watson

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We have cloned and expressed recombinant guinea pig tumor necrosis factor-α (gpTNF-α) and examined its inflammatory activities after tracheal instillation in guinea pigs. A 1, 071-bp cDNA, including the region encoding the full-length 234-amino acid gpTNF-α protein, was cloned from concanavalin A-stimulated guinea pig splenocytes. The 154-amino acid protein corresponding to secreted gpTNF-α was expressed as a fusion protein in Escherichia coli, purified by affinity chromatography, and cleaved to yield a 17-kDa protein. gpTNF-α had a cytotoxic effect on WEHI 164 cells and was detected by goat anti-murine tumor necrosis factor-α (TNF-α) antibody in Western blots. Intratracheal instillation of gpTNF-α (50-150 ng) caused pronounced and dose-dependent airway eosinophilia. Incubation of gpTNF-α with rabbit anti-murine TNF-α sera or heating the gpTNF-α before instillation reduced bronchoalveolar lavage (BAL) eosinophils to near control levels. Maximum BAL eosinophilia was observed at 24 h, but eosinophil numbers remained significantly above vehicle-treated animals for 72 h. Hence, gpTNF-a elicits a pronounced and protracted eosinophil accumulation in the guinea pig lung.

Original languageEnglish
Pages (from-to)G735-G747
JournalAmerican Journal of Physiology
Volume273
Issue number3 PART 1
Publication statusPublished - Dec 1 1997
Externally publishedYes

Keywords

  • Cytokine
  • Eosinophil
  • Lung inflammation

ASJC Scopus subject areas

  • Physiology (medical)

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  • Cite this

    White, A. M., Yoshimura, T., Smith, A. W., Westwick, J., & Watson, M. L. (1997). Airway inflammation induced by recombinant guinea pig tumor necrosis factor-α. American Journal of Physiology, 273(3 PART 1), G735-G747.