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

28 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-hp 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-α elicits a pronounced and protracted eosinophil accumulation in the guinea pig lung.

Original languageEnglish
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume273
Issue number3 17-3
Publication statusPublished - Sep 1997
Externally publishedYes

Fingerprint

Guinea Pigs
Tumor Necrosis Factor-alpha
Inflammation
Eosinophils
Eosinophilia
Bronchoalveolar Lavage
Amino Acids
Proteins
Escherichia coli Proteins
Concanavalin A
Affinity Chromatography
Goats
Heating
Complementary DNA
Western Blotting
Rabbits
Lung
Antibodies
Serum

Keywords

  • Cytokine
  • Eosinophil
  • Lung inflammation

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology

Cite this

Airway inflammation induced by recombinant guinea pig tumor necrosis factor-α. / White, Anna Marie; Yoshimura, Teizo; Smith, Anthony W.; Westwick, John; Watson, Malcolm L.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 273, No. 3 17-3, 09.1997.

Research output: Contribution to journalArticle

White, Anna Marie ; Yoshimura, Teizo ; Smith, Anthony W. ; Westwick, John ; Watson, Malcolm L. / Airway inflammation induced by recombinant guinea pig tumor necrosis factor-α. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 1997 ; Vol. 273, No. 3 17-3.
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