Enhanced Characterization of Serum Autoantibody Reactivity Following HSP 60 Immunization in a Rat Model of Experimental Autoimmune Glaucoma
Enhanced Characterization of Serum Autoantibody Reactivity Following HSP 60 Immunization in a Rat Model of Experimental Autoimmune Glaucoma
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DOI：
10.3109/02713683.2010.495829
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发表时间：
2010-10-01
影响因子：
2
通讯作者：
Grus, F. H.
中科院分区：
文献类型：
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作者：
Joachim, S. C.;Wax, M. B.;Grus, F. H.
目的：在几项研究中，已经在人青光眼患者的血清中鉴定出针对热休克蛋白的抗体，并且在实验性自身免疫性青光眼的动物模型中，用热休克蛋白60（HSP 60）免疫导致视网膜神经节细胞（RGC）损失。本研究的目的是观察血清中抗视网膜抗体增加的时间过程，并表征识别伴随HSP 60免疫大鼠模型的实验性自身免疫性glaucoma.Methods的突出自身抗体：免疫后8周HSP 60视网膜flatmounts制备和RGCs计数在8个预定义的领域，并与对照组相比。免疫前以及免疫后4周和8周收集的血清用于使用Western印迹技术检测针对牛视网膜抗原的抗体模式。这些模式进行了分析，多元统计方法。通过质谱法进一步鉴定显著增加的自身抗体。结果：8周后，HSP 60免疫的动物表现出显著的视网膜平坦区RGC丢失（P = 0.02），这是眼内压无关的。检测到抗体谱的早期变化，其中许多显著上调。四周后抗体反应性显著上调的抗原被鉴定为组蛋白H2 B 1型、S-抑制蛋白、胶质细胞酸性蛋白、波形蛋白和热休克蛋白60。这些上调的自身抗体在其初始上调后四周恢复到正常水平。另一方面，抗体对视网膜醛结合蛋白1成为上调后8 weeks.Conclusion：这是第一次研究，以确定视网膜自身抗体的出现和消失在大鼠血清中的几个时间点后，他们的初始上调响应HSP 60免疫的实验性自身免疫性青光眼模型。
Purpose: Antibodies against heat shock proteins have been identified in sera of human glaucoma patients in several studies and immunization with heat shock protein 60 (HSP 60) causes retinal ganglion cell (RGC) loss in an animal model of experimental autoimmune glaucoma. The aim of this study was to observe the time course of increased anti-retina antibody appearance in the serum and characterize the identification of prominent autoantibodies that accompany HSP 60 immunization in a rat model of experimental autoimmune glaucoma.Methods: Eight weeks after immunization with HSP 60 retinal flatmounts were prepared and RGCs were counted in eight predefined areas and compared to controls. Serum collected before, as well as four and eight weeks after, immunization was used to detect antibody patterns against bovine retinal antigens using Western blotting techniques. These patterns were analyzed by multivariate statistical methods. Autoantibodies that were prominently increased were further identified through mass spectrometry. Intraocular pressure was measured throughout the study.Results: After eight weeks, animals immunized with HSP 60 showed significant RGC loss of retinal flatmounts (P = 0.02), which was intraocular pressure independent. Early changes in antibody profiles, many of them significant upregulations, were detected. Antigens with significantly upregulated antibody reactivity after four weeks were identified as histone H2B type 1, S-arrestin, glial fibrillary acidic protein, vimentin, and heat shock protein 60. These upregulated autoantibodies returned to normal levels four weeks following their initial upregulation. Antibodies against retinaldehyde binding protein 1 on the other hand became upregulated after eight weeks.Conclusion: This is the first study to identify the appearance and disappearance of retinal autoantibodies in the serum of rats at several time-points following their initial upregulation in response to HSP 60 immunization in a model of experimental autoimmune glaucoma.