Potentially antigenic RBC membrane proteins in dogs with primary immune-mediated hemolytic anemia
Potentially antigenic RBC membrane proteins in dogs with primary immune-mediated hemolytic anemia
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DOI：
10.1111/j.1939-165x.2011.00391.x
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发表时间：
2012-03-01
影响因子：
1.2
通讯作者：
Wood, Darren
中科院分区：
文献类型：
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作者：
Tan, Emmeline;Bienzle, Dorothee;Wood, Darren
背景：原发性免疫介导的溶血性贫血（IMHA）是犬发病和死亡的重要原因。自身免疫反应性的机制仍然知之甚少。Objective：本研究的目的是确定红细胞膜蛋白，可能是抗原性的狗与原发性IMHA.Methods：用二甲苯洗脱抗体从红细胞的12只狗IMHA，4只狗贫血，由于其他原因比IMHA，和2只健康狗。从17只健康狗的血液中制备合并的RBC膜蛋白。然后通过免疫印迹分析洗脱的抗体与合并的膜蛋白和自体血浆的相互作用。带存在于12只狗IMHA，但不是在其他6只狗被认为是潜在的自身抗原，并确定了液相色谱串联mass spectrometry.Results：从所有18只狗的红细胞洗脱液对带3蛋白的反应性。在患有IMHA的犬中唯一鉴定了6种其他蛋白质的抗体。钙蛋白酶，补体成分3，和过氧化物酶2的反应被确定在8，8，和4的12个样品，分别从狗IMHA，但没有从6只狗没有IMHA.Conclusions的样品：检测普遍免疫反应对带3蛋白可能表明识别衰老红细胞。在患有IMHA的犬中，抗体唯一识别的蛋白质参与氧化应激和细胞凋亡（钙蛋白酶）、炎症（补体）和活性氧（过氧化物氧还蛋白2）的清除。这些蛋白质在引发自身免疫中是否重要，或者IMHA期间是否产生靶向这些蛋白质的免疫球蛋白，仍有待确定。
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 RBCs that could be antigenic in dogs with primary IMHA.Methods: Antibodies were eluted with xylene from RBCs 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.