Generation of a mouse tumor necrosis factor mutant with antiperitonitis and desensitization activities comparable to those of the wild type but with reduced systemic toxicity.

TitleGeneration of a mouse tumor necrosis factor mutant with antiperitonitis and desensitization activities comparable to those of the wild type but with reduced systemic toxicity.
Publication TypeJournal Article
Year of Publication1997
AuthorsLucas, R., B. Echtenacher, E. Sablon, P. Juillard, S. Magez, J. Lou, Y. Donati, F. Bosman, A. Van de Voorde, L. Fransen, D. N. Männel, G. E. Grau, and P. De Baetselier
JournalInfect Immun
Volume65
Issue6
Pagination2006-10
Date Published1997 Jun
ISSN0019-9567
KeywordsAnimals, Lectins, Male, Mice, Mice, Inbred C57BL, Mutation, Peritonitis, Receptors, Tumor Necrosis Factor, Tumor Necrosis Factor-alpha
Abstract

In this study, we investigated whether the recently identified lectin-like domain of tumor necrosis factor (TNF) is implicated in its biological activities on mammalian cells. To this end, a mouse TNF (mTNF) triple mutant, T104A-E106A-E109A mTNF (referred to hereafter as triple mTNF), lacking the lectin-like affinity of mTNF for specific oligosaccharides, was compared with the wild-type molecule for various TNF effects in vitro and in vivo. The triple mTNF displayed a 50-fold-reduced TNF receptor 2 (TNFR2)-mediated bioactivity but only a 5-fold-reduced TNFR1-mediated bioactivity in vitro. The specific activity of the triple mutant on L929 fibrosarcoma cells was slightly reduced compared with that of the wild type. We subsequently assessed the systemic toxicity of triple versus wild-type mTNF, since TNFR2 is partially implicated in this activity. The triple mTNF had a significantly reduced toxicity compared with that of wild-type mTNF in vivo. Moreover, we compared the effects of the triple and the wild-type mTNFs in TNFR1-mediated phenomena, such as (i) induction of tolerance towards a lethal mTNF dose and (ii) protective activity in cecal ligation and puncture-induced septic peritonitis. No significant differences between the mutant and wild-type forms were observed. In conclusion, these results indicate that triple mTNF, lacking TNF's lectin-like binding capacity, has reduced systemic toxicity but retains the tolerance-inducing and peritonitis-protective activities of wild-type mTNF.

Alternate JournalInfect. Immun.
PubMed ID9169725
PubMed Central IDPMC175277