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首页> 外文期刊>Acta Neuropathologica >Differentiation of prion protein glycoforms from naturally occurring sheep scrapie, sheep-passaged scrapie strains (CH1641 and SSBP1), bovine spongiform encephalopathy (BSE) cases and Romney and Cheviot breed sheep experimentally inoculated with BSE using two monoclonal antibodies
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Differentiation of prion protein glycoforms from naturally occurring sheep scrapie, sheep-passaged scrapie strains (CH1641 and SSBP1), bovine spongiform encephalopathy (BSE) cases and Romney and Cheviot breed sheep experimentally inoculated with BSE using two monoclonal antibodies

机译：two病毒蛋白糖型与天然羊瘙痒病，传代绵羊瘙痒病菌株（CH1641和SSBP1），牛海绵状脑病（BSE）病例以及使用两种单克隆抗体实验接种BSE的罗姆尼和雪维特种羊的区别

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A panel of ruminant brain tissues were subjected to a Western immunoblotting technique using two monoclonal antibodies (mAbs). The resultant prion protein (PrP) glycoforms showed that three distinctions can be made between natural ovine scrapie cases and sheep experimentally inoculated with bovine spongiform encephalopathy (BSE). Differentiation between BSE-infected cattle and natural cases of sheep scrapie was also possible using these two antibodies. There were subtle differences in the molecular weight positions of the di-glycosylated, mono-glycosylated and unglycosylated forms of the abnormal PrP (PrPSc) associated with these ruminant transmissible spongiform encephalopathies. In particular, a distinct difference for the unglycosylated protein band was observed. For ovine scrapie samples, this band was noticeably of a higher molecular weight than that found for brain samples from the Romney and Cheviot breed sheep infected with BSE and, to a lesser degree, higher than that observed for bovine BSE samples. Using the comparison of glycoform ratios, the technique provided a distinction between the sheep experimentally infected with BSE and natural cases of sheep scrapie but did not provide a distinction between natural cases of bovine BSE and ovine scrapie. The sheep-passaged CH1641 scrapie strain gave molecular weights similar to, but not identical to BSE, and a glycoform ratio similar to ovine scrapie cases. The SSBP1 experimental scrapie strain gave molecular weights that were akin to natural scrapie cases but the glycoform ratio was different to that found for all the other samples. When mAb P4 was substituted for mAb 6H4 in the technique, only the natural scrapie samples and SSBP1 gave strong signals. BSE in sheep and the CH1641 strain gave weak reactions and PrPSc from BSE-infected cattle could not be detected at all. The results suggest that this combination of molecular weight and glycoform ratio analyses, and differentiation with two specific antibodies could be used to provide a possible screening test for BSE in the UK sheep flock, if confirmed as accurate by bioassay and lesion profile analysis in mice inoculated with brain tissue from suspect field cases.
机译：使用两个单克隆抗体（mAb）对一组反刍动物的脑组织进行了Western免疫印迹技术。产生的病毒蛋白（PrP）糖型显示，天然羊瘙痒病病例与实验接种牛海绵状脑病（BSE）的绵羊之间可以有三个区别。使用这两种抗体，也有可能在BSE感染的牛和羊瘙痒病的自然病例之间进行区分。与反刍动物可传播的海绵状脑病相关的异常PrP（PrPSc ）的二糖基化，单糖基化和未糖基化形式的分子量位置存在细微差异。特别地，观察到未糖基化蛋白带的明显差异。对于绵羊瘙痒病样品，该条带的分子量明显高于感染BSE的罗姆尼和Cheviot品种绵羊的脑部样品的分子量，并且在较小程度上高于牛BSE样品的分子量。通过比较糖型比率，该技术在实验感染BSE的绵羊和羊瘙痒病的自然病例之间进行了区分，但没有在牛BSE和羊瘙痒症的自然病例之间进行区分。绵羊传代的CH1641瘙痒病菌株的分子量与BSE相似但不相同，而糖型比与绵羊瘙痒病病例相似。 SSBP1实验性瘙痒病菌株的分子量类似于自然瘙痒病病例，但糖型比不同于所有其他样品的分子量。当在该技术中用mAb P4代替mAb 6H4时，只有天然的瘙痒病样品和SSBP1发出强信号。绵羊中的疯牛病和CH1641株反应较弱，根本无法检测到被疯牛病感染的牛的PrPSc 。结果表明，这种分子量和糖型比率分析的组合，以及与两种特异性抗体的区分，可以被用来为英国绵羊群中的疯牛病提供可能的筛查试验，如果通过生物测定和病灶分布分析在接种的小鼠中被证实是准确的话带有可疑野外病例的脑组织。

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