Am. White et al., AIRWAY INFLAMMATION-INDUCED BY RECOMBINANT GUINEA-PIG TUMOR-NECROSIS-FACTOR-ALPHA, American journal of physiology. Lung cellular and molecular physiology, 17(3), 1997, pp. 524-530
We have cloned and expressed recombinant guinea pig tumor necrosis fac
tor-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, wa
s cloned from concanavalin A-stimulated guinea pig splenocytes. The 15
4-amino acid protein corresponding to secreted gpTNF-alpha was express
ed as a fusion protein in Escherichia coli, purified by affinity chrom
atography, and cleaved to yield a 17-kDa protein. gpTNF-alpha had a cy
totoxic effect on WEHI 164 cells and was detected by goat anti-murine
tumor necrosis factor-alpha (TNF-alpha) antibody in Western blots. Int
ratracheal instillation of gpTNF-alpha (50-150 ng) caused pronounced a
nd dose-dependent airway eosinophilia. Incubation of gpTNF-alpha with
rabbit anti-murine TNF-alpha sera or heating the gpTNF-alpha before in
stillation reduced bronchoalveolar lavage (BAL) eosinophils to near co
ntrol levels. Maximum BAL eosinophilia was observed at 24 h, but eosin
ophil numbers remained significantly above vehicle-treated animals for
72 h. Hence, gpTNF-alpha elicits a pronounced and protracted eosinoph
il accumulation in the guinea pig lung.