Retrocyclin reinforcement of pulmonary defenses against viral aerosols

简介：除了人类白细胞和上皮细胞产生的防御素和防御素外，许多非人灵长类动物也产生？-防御蛋白，代表第三个防御蛋白亚家族。？-防御素缺乏有效的抗菌或抗真菌活性，但具有广泛的抗病毒特性。反转录细胞周期素是什么？-结构基于表达（但未翻译）人类基因序列的防御蛋白？-defensin伪基因。广泛的长期目标：使用反转录细胞周期蛋白来增强肺部对病毒气溶胶的防御。具体的目标。1. 测试逆转录细胞周期蛋白对潜在的病毒生物恐怖主义制剂的作用，包括甲型流感和乙型流感（正粘病毒科）、HPIV3（副粘病毒科）、西尼罗河和黄热病病毒（黄病毒科）、sars冠状病毒（冠状病毒科）、痘苗（痘病毒科）、VEE、甲病毒（托加病毒科）和塔卡瑞病毒（沙粒病毒科）。观察反转录细胞周期蛋白与表面活性蛋白SP-A和SP-D的相互作用。3. 目的：观察RC2对肺表面活性物质磷脂单层和双层的影响。研究设计。逆转录细胞周期素-2 （RC-2）和选定的类似物将通过固相肽合成制备。病毒将通过标准的空斑减少和CPE减少试验进行体外测试。反转录细胞周期蛋白对表面活性剂磷脂膜完整性的影响将在单层系统中通过Langmuir-Blodgett槽进行测试，在双层系统中通过由表面活性剂脂质和相关表面活性剂蛋白SP-B和SP-C组成的囊泡进行测试。反转录细胞周期蛋白与SP-A和SP-D的结合将通过表面等离子体共振进行研究。反转录细胞周期素对SP-A和SP-D内在抗病毒活性的影响（反之亦然）将在标准斑块减少和CPE减少试验中使用从屠宰场获得的猪肺中纯化的蛋白质进行研究。保健关系和机构使命。本申请是对pa04 -119的直接回应。外行语言总结。许多自然病毒感染始于接触含有病毒的空气。病毒气雾剂也可能让生物恐怖分子传播致残病毒。我们建议测试逆转录细胞周期蛋白（新发现的抗病毒肽）对潜在的病毒生物恐怖主义制剂的活性，并确定它们与覆盖肺气囊表面的薄流体层elf的生物相容性。

DESCRIPTION (provided by applicant): Introduction: In addition to the a and ¿ defensins produced by human leukocytes and epithelial cells, many nonhuman primates produce ?-defensins, representing a third defensin subfamily. ?-defensins lack potent antibacterial or antifungal activity, but have broad antiviral properties. Retrocyclins are ?-defensins whose structures are based on the sequences of expressed (but untranslated) human ?-defensin pseudogenes. Broad, Long term objectives: To use retrocyclins to enhance pulmonary defenses to viral aerosols. Specific aims. 1. To test retrocyclins against potential viral bioterrorism agents, including Influenza A and B (Orthomyxoviridae), HPIV3 (Paramyxoviridae), West Nile and Yellow Fever virus (Flaviviridae) SARS-CoV (Coronaviridae), Vaccinia (Poxviridae), VEE, an alphavirus (Togaviridae), and Tacaribe virus (Arenaviridae) 2. To examine the interactions of retrocyclins with surfactant proteins SP-A and SP-D. 3. To test the effects actions of RC2 on monolayers and bilayers of pulmonary surfactant phospholipids. Research Design. Retrocyclin-2 (RC-2) and selected analogs will be prepared by solid phase peptide synthesis. Viruses will be tested in vitro by standard plaque reduction and CPE reduction assays. The effects of retrocyclins on the integrity of surfactant phospholipid films will be tested in monolayer systems by using a Langmuir-Blodgett trough, and in bilayer systems by using vesicles composed of surfactant lipids and associated surfactant proteins SP-B and SP-C. Binding of retrocyclins to SP-A and SP-D will be studied by surface plasmon resonance. Effects of retrocyclins on the intrinsic antiviral activity of SP-A and SP-D (and vice versa) will be studied using proteins purified from abattoir-obtained porcine lungs in standard plaque reduction and CPE reduction assays. Health relatedness and Agency Mission. This application is in direct response to PA 04-119. Lay language summary. Many natural viral infections begin with exposure to air that contains viruses. Viral aerosols may also allow bioterrorists to deliver incapacitating viruses. We propose to test the activity of retrocyclins (newly discovered antiviral peptides) against potential viral bioterrorism agents, and to determine their biocompatability with ELF-the thin fluid layer that covers the surface of the pulmonary air sacs.