Kamishina Hiroaki, Oji Takashi, Cheeseman Jennifer A, Clemmons Roger M
Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL, USA.
Vet Clin Pathol. 2008 Jun;37(2):217-20. doi: 10.1111/j.1939-165X.2008.00020.x.
Detection of intrathecal IgG synthesis is important in evaluating inflammatory diseases in the central nervous system. Isoelectric focusing (IEF) is currently the most sensitive method to demonstrate intrathecal IgG synthesis and may have diagnostic value for German Shepherd degenerative myelopathy (GSDM).
The objective of this study was to adapt a modified IEF and immunofixation method for the detection of oligoclonal bands in cerebrospinal fluid (CSF) from dogs with GSDM.
Serum and lumbar CSF samples were collected from 6 German Shepherd dogs clinically diagnosed with GSDM. Samples were also collected from 6 clinically healthy dogs for comparison. The concentration of IgG was measured by quantitative ELISA and the concentration of total protein was measured by the Bradford protein assay. The presence of oligoclonal bands was evaluated by use of modified IEF followed by immunofixation.
The concentrations of total protein and IgG, and the IgG/total protein ratio in CSF samples, were not significantly different between GSDM patients and control dogs. Four GSDM patients had oligoclonal bands in their CSF based on IEF-immunofixation. No oligoclonal bands were found in CSF from control dogs.
The results suggest that the detection of oligoclonal bands by IEF-immunofixation may have diagnostic value for GSDM, and support the idea that humoral immune responses may play a role in the pathogenesis of GSDM.
鞘内IgG合成的检测对于评估中枢神经系统的炎症性疾病很重要。等电聚焦（IEF）是目前证明鞘内IgG合成最敏感的方法，可能对德国牧羊犬退行性脊髓病（GSDM）具有诊断价值。
本研究的目的是采用改良的IEF和免疫固定方法检测GSDM犬脑脊液（CSF）中的寡克隆带。
从6只临床诊断为GSDM的德国牧羊犬采集血清和腰椎CSF样本。还从6只临床健康犬采集样本用于比较。通过定量ELISA测定IgG浓度，通过Bradford蛋白测定法测定总蛋白浓度。采用改良的IEF随后进行免疫固定来评估寡克隆带的存在。
GSDM病犬和对照犬CSF样本中的总蛋白和IgG浓度以及IgG/总蛋白比值无显著差异。基于IEF-免疫固定，4只GSDM病犬的CSF中有寡克隆带。对照犬的CSF中未发现寡克隆带。
结果表明，IEF-免疫固定检测寡克隆带可能对GSDM具有诊断价值，并支持体液免疫反应可能在GSDM发病机制中起作用的观点。
