Neutrophils from patients with TNFRSF1A mutations display resistance to tumor necrosis factor-induced apoptosis: pathogenetic and clinical implications.
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.
D'Osualdo
Andrea
A
Istituto G. Gaslini, Genoa, Italy.
Ferlito
Francesca
F
Prigione
Ignazia
I
Obici
Laura
L
Meini
Antonella
A
Zulian
Francesco
F
Pontillo
Alessandra
A
Corona
Fabrizia
F
Barcellona
Roberto
R
Di Duca
Marco
M
Santamaria
Giuseppe
G
Traverso
Francesco
F
Picco
Paolo
P
Baldi
Maurizia
M
Plebani
Alessandro
A
Ravazzolo
Roberto
R
Ceccherini
Isabella
I
Martini
Alberto
A
Gattorno
Marco
M
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*
D017304:Annexin A5; D047888:Receptors, Tumor Necrosis Factor, Type I; D014409:Tumor Necrosis Factor-alpha; C505530:CASP8 protein, human; D053181:Caspase 8; D020169:Caspases