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Potentially antigenic RBC membrane proteins in dogs with primary immune‐mediated hemolytic anemia
Author(s) -
Tan Emmeline,
Bienzle Dorothee,
Shewen Patricia,
Kruth Stephen,
Wood Darren
Publication year - 2012
Publication title -
veterinary clinical pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.537
H-Index - 51
eISSN - 1939-165X
pISSN - 0275-6382
DOI - 10.1111/j.1939-165x.2011.00391.x
Subject(s) - immunology , antibody , complement system , antigen , immune system , hemolytic anemia , haptoglobin , epitope , membrane protein , biology , chemistry , biochemistry , membrane
Background Primary immune‐mediated hemolytic anemia ( IMHA ) is an important cause of morbidity and mortality in dogs. The mechanisms underlying autoimmune reactivity remain poorly understood. Objective The aim of this study was to identify membrane proteins of RBC s that could be antigenic in dogs with primary IMHA . Methods Antibodies were eluted with xylene from RBC s of 12 dogs with IMHA , 4 dogs with anemia due to causes other than IMHA , and 2 healthy dogs. Pooled RBC membrane proteins were prepared from blood of 17 healthy dogs. The eluted antibodies were then analyzed by immunoblotting for interactions with the pooled membrane proteins and autologous plasma. Bands present in the 12 dogs with IMHA but not in the 6 other dogs were considered potential autoantigens and were identified by liquid chromatography followed by tandem mass spectrometry. Results RBC eluates from all 18 dogs had reactivity against band 3 protein. Antibodies to 6 additional proteins were uniquely identified in dogs with IMHA . Reactivity to calpain, complement component 3, and peroxiredoxin 2 was identified in 8, 8, and 4 of the 12 samples, respectively, from dogs with IMHA , but in none of the samples from the 6 dogs without IMHA . Conclusions Detection of universal immune reactivity against band 3 protein probably indicates recognition of senescent RBC . Proteins uniquely recognized by antibodies in dogs with IMHA are involved in oxidative stress and apoptosis (calpain), inflammation (complement), and scavenging of reactive oxygen species (peroxiredoxin 2). It remains to be determined if these proteins are important in initiating autoimmunity or if immunoglobulins targeting these proteins develop during IMHA .

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