title: Airway inflammation induced by recombinant guinea pig tumor necrosis factor-alpha.
abstract: We have cloned and expressed recombinant guinea pig tumor necrosis factor-alpha (gpTNF-alpha) 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-alpha protein, was cloned from concanavalin A-stimulated guinea pig splenocytes. The 154-amino acid protein corresponding to secreted gpTNF-alpha was expressed as a fusion protein in Escherichia coli, purified by affinity chromatography, and cleaved to yield a 17-kDa protein. gpTNF-alpha had a cytotoxic effect on WEHI 164 cells and was detected by goat anti-murine tumor necrosis factor-alpha (TNF-alpha) antibody in Western blots. Intratracheal instillation of gpTNF-alpha (50-150 ng) caused pronounced and dose-dependent airway eosinophilia. Incubation of gpTNF-alpha with rabbit anti-murine TNF-alpha sera or heating the gpTNF-alpha 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-alpha elicits a pronounced and protracted eosinophil accumulation in the guinea pig lung.
authors.affiliation: School of Pharmacy and Pharmacology, University of Bath, United Kingdom.
authors.forename: A M
authors.initials: AM
authors.lastname: White
authors.forename: T
authors.initials: T
authors.lastname: Yoshimura
authors.forename: A W
authors.initials: AW
authors.lastname: Smith
authors.forename: J
authors.initials: J
authors.lastname: Westwick
authors.forename: M L
authors.initials: ML
authors.lastname: Watson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001483:Base Sequence; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D003001:Cloning, Molecular; D017931:DNA Primers; D004804:Eosinophils / Q000187:drug effects / Q000502:physiology; D004926:Escherichia coli; D005260:Female; D006168:Guinea Pigs; D006801:Humans; D007249:Inflammation; D007962:Leukocytes / Q000187:drug effects / Q000502:physiology*; D008168:Lung / Q000187:drug effects* / Q000503:physiopathology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000276:immunology; D013997:Time Factors; D014132:Trachea; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D000906:Antibodies; D017931:DNA Primers; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha