title: 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.
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.
authors.affiliation: Department of Anesthesiology, Pharmacology and Surgical Intensive Care, University Medical Center, University of Geneva, Switzerland.
authors.forename: R
authors.initials: R
authors.lastname: Lucas
authors.forename: B
authors.initials: B
authors.lastname: Echtenacher
authors.forename: E
authors.initials: E
authors.lastname: Sablon
authors.forename: P
authors.initials: P
authors.lastname: Juillard
authors.forename: S
authors.initials: S
authors.lastname: Magez
authors.forename: J
authors.initials: J
authors.lastname: Lou
authors.forename: Y
authors.initials: Y
authors.lastname: Donati
authors.forename: F
authors.initials: F
authors.lastname: Bosman
authors.forename: A
authors.initials: A
authors.lastname: Van de Voorde
authors.forename: L
authors.initials: L
authors.lastname: Fransen
authors.forename: D N
authors.initials: DN
authors.lastname: Männel
authors.forename: G E
authors.initials: GE
authors.lastname: Grau
authors.forename: P
authors.initials: P
authors.lastname: de Baetselier
mesh_terms: D000818:Animals; D037102:Lectins / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D010538:Peritonitis / Q000517:prevention & control*; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism / Q000627:therapeutic use* / Q000633:toxicity
chemical_list: D037102:Lectins; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha