title: Neutrophils from patients with TNFRSF1A mutations display resistance to tumor necrosis factor-induced apoptosis: pathogenetic and clinical implications.
abstract: OBJECTIVE
  To explore tumor necrosis factor (TNF)-induced apoptosis in neutrophils from patients with TNF receptor-associated periodic syndrome (TRAPS) and to correlate the results with the different kinds of TNFRSF1A mutations.
  
  
  METHODS
  Two hundred sixty-five patients with clinically suspected inherited autoinflammatory syndrome were screened for mutations of the TNFRSF1A gene. Neutrophils were isolated from heparinized blood by dextran sedimentation and incubated with and without cycloheximide (CHX) and TNFalpha. Cell apoptosis was assessed by human annexin V binding, and caspase 8 activation was assessed by flow cytometry.
  
  
  RESULTS
  Twenty-one patients were found to carry a variant of the TNFRSF1A gene: 13 patients had an R92Q substitution, and 8 patients presented other missense substitutions, 1 splicing mutation, and 1 in-frame interstitial deletion. Neutrophil stimulation with TNF and CHX was associated with induction of apoptosis in 12 normal controls and in 10 subjects with the R92Q mutation. Conversely, neutrophils from 8 TRAPS patients with mutations of cysteine or threonine residues or interstitial deletion did not show any induction of apoptosis after stimulation. The incidence of the R92Q mutation among patients with recurrent autoinflammatory syndromes was similar to that observed in the normal population.
  
  
  CONCLUSIONS
  Resistance to TNF-mediated apoptosis is a feature in TRAPS patients who have mutations of cysteine residues or interstitial deletion, and may play a pathogenic role. The R92Q mutation does not appear to be significantly associated with TRAPS.
authors.affiliation: Istituto G. Gaslini, Genoa, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: D'Osualdo
authors.forename: Francesca
authors.initials: F
authors.lastname: Ferlito
authors.forename: Ignazia
authors.initials: I
authors.lastname: Prigione
authors.forename: Laura
authors.initials: L
authors.lastname: Obici
authors.forename: Antonella
authors.initials: A
authors.lastname: Meini
authors.forename: Francesco
authors.initials: F
authors.lastname: Zulian
authors.forename: Alessandra
authors.initials: A
authors.lastname: Pontillo
authors.forename: Fabrizia
authors.initials: F
authors.lastname: Corona
authors.forename: Roberto
authors.initials: R
authors.lastname: Barcellona
authors.forename: Marco
authors.initials: M
authors.lastname: Di Duca
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Santamaria
authors.forename: Francesco
authors.initials: F
authors.lastname: Traverso
authors.forename: Paolo
authors.initials: P
authors.lastname: Picco
authors.forename: Maurizia
authors.initials: M
authors.lastname: Baldi
authors.forename: Alessandro
authors.initials: A
authors.lastname: Plebani
authors.forename: Roberto
authors.initials: R
authors.lastname: Ravazzolo
authors.forename: Isabella
authors.initials: I
authors.lastname: Ceccherini
authors.forename: Alberto
authors.initials: A
authors.lastname: Martini
authors.forename: Marco
authors.initials: M
authors.lastname: Gattorno
mesh_terms: D000328:Adult; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology*; D053181:Caspase 8; D020169:Caspases / Q000494:pharmacology; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D008297:Male; D009154:Mutation; D009504:Neutrophils / Q000502:physiology*; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000235:genetics*; D013577:Syndrome; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*
chemical_list: D017304:Annexin A5; D047888:Receptors, Tumor Necrosis Factor, Type I; D014409:Tumor Necrosis Factor-alpha; C505530:CASP8 protein, human; D053181:Caspase 8; D020169:Caspases