Mutant tumor necrosis factor receptor associated with tumor necrosis factor receptor-associated periodic syndrome is altered antigenically and is retained within patients' leukocytes.
OBJECTIVE
To investigate the effect of mutations in tumor necrosis factor receptor superfamily member 1A (TNFRSF1A) in TNFR-associated periodic syndrome (TRAPS) on the binding of anti-TNFRSF1A monoclonal antibodies (mAb), and to investigate the subcellular distribution of mutant versus wild-type (WT) TNFRSF1A in patients with TRAPS.


METHODS
HEK 293 cells transfected with WT and/or mutant TNFRSF1A were used to investigate the interaction of anti-TNFRSF1A mAb with the WT and mutant proteins. Monoclonal antibodies that differentially bound to C33Y TNFRSF1A were used to investigate the distribution of WT and mutant TNFRSF1A in TRAPS patients with the C33Y mutation.


RESULTS
We identified a mAb whose binding to TNFRSF1A was completely abolished by the C33Y or C52F TRAPS-associated mutations, whereas other mutations (T50M, C88Y, R92Q) had lesser effects on the binding of this mAb. A different mAb was found to bind efficiently to all of the mutant forms of TNFRSF1A examined as well as to the WT receptor. Exploitation of the differential binding properties of these mAb indicated that mutant (as distinct from WT) TNFRSF1A showed abnormal intracellular retention in the neutrophils of TRAPS patients with the C33Y mutation, with little if any expression of mutant TNFRSF1A on the cell surface or as soluble receptor in plasma.


CONCLUSIONS
TRAPS-associated mutant TNFRSF1A has an antigenically altered structure and shows abnormal retention in the leukocytes of patients with TRAPS, which is consistent with previous findings from in vitro and transgenic model systems. This is consistent with a misfolded protein response contributing to the pathophysiology of TRAPS.
Todd
Ian
I
Division of Immunology, Queen's Medical Center, University of Nottingham, Derby Road, Nottingham, UK. ian.todd@nottingham.ac.uk
Radford
Paul M
PM
Daffa
Noura
N
Bainbridge
Susan E
SE
Powell
Richard J
RJ
Tighe
Patrick J
PJ
D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D000916:Antibody Diversity / Q000235:genetics* / Q000276:immunology; D001665:Binding Sites / Q000235:genetics / Q000276:immunology; D002460:Cell Line; D010505:Familial Mediterranean Fever / Q000235:genetics* / Q000276:immunology; D006801:Humans; D009154:Mutation; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism*; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014162:Transfection
D000911:Antibodies, Monoclonal; D047888:Receptors, Tumor Necrosis Factor, Type I; C506235:TNFRSF1A protein, human