Structures and Activities of Prions and Prion Proteins

朊病毒和朊病毒蛋白的结构和活性

• 批准号: 8555782
• 负责人: BYRON CAUGHEY
• 金额: $ 44.65万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8555782
• 关键词: Antibodies; Apoptosis; Binding Proteins; Bovine Spongiform Encephalopathy; Brain; Cells; Central Nervous System Infections; Chronic Wasting Disease; Complement; Complement 1q; Creutzfeldt-Jakob Syndrome; Deposition; Disease; Goals; Immunohistochemistry; In Vitro; Infection; Molecular; Mus; Neurodegenerative Disorders; Pathogenesis; Phosphatidylserines; PrPSc Proteins; Prion Diseases; Prions; Role; Scrapie; Serum; Staining method; Stains; Structure; annexin A5; disease transmission; in vivo; transmission process

Accomplishments for FY2012: We have identified scrapie strain-dependent interactions of PrPSc with complement factors in vitro and in vivo. Roles of complement factors in prion infection of the central nervous system remain unclear. In this study, we assessed the strain-dependent reactivity of complement factors in prion infections of Neuro2a (N2a) cells and mouse brains. N2a cells persistently infected with either Chandler or 22L scrapie strains were cultured in the presence of normal mouse serum (NMS), followed by staining with the phosphatidylserine binding protein and early apoptosis marker Annexin V. The proportion of Annexin V positive cells was increased both in Chandler- and 22L-infected cells. Preincubation of NMS with anti-C1q, C3 and/or C9 antibodies reduced Annexin V positive cells in Chandler-infected cells, while only anti-C3 antibodies were effective on 22L-infected cells. Immunohistochemistry showed that deposition of C1q and C3 was different between Chandler- and 22L-infected mouse brains. These results indicate that the reactivity of complement factors differs between prion strains both in vitro and in vivo.

2012 财年的成就：我们在体外和体内鉴定了 PrPSc 与补体因子的痒病菌株依赖性相互作用。补体因子在中枢神经系统朊病毒感染中的作用仍不清楚。在这项研究中，我们评估了 Neuro2a (N2a) 细胞和小鼠大脑的朊病毒感染中补体因子的菌株依赖性反应性。将持续感染 Chandler 或 22L 痒病菌株的 N2a 细胞在正常小鼠血清 (NMS) 存在下培养，然后用磷脂酰丝氨酸结合蛋白和早期凋亡标记膜联蛋白 V 染色。钱德勒和 22L 感染的细胞中膜联蛋白 V 阳性细胞的比例均增加。 NMS 与抗 C1q、C3 和/或 C9 抗体预孵育可减少 Chandler 感染细胞中的膜联蛋白 V 阳性细胞，而仅抗 C3 抗体对 22L 感染细胞有效。免疫组织化学显示，Chandler 和 22L 感染的小鼠大脑中 C1q 和 C3 的沉积不同。这些结果表明，在体外和体内，朊病毒株之间的补体因子的反应性不同。