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外，许多非人类灵长类动物还产生？-防御素，代表第三个防御素亚家族。？-防御素缺乏强大的抗细菌或抗真菌活性，但具有广泛的抗病毒特性。逆转录周期蛋白是一种？-防御素，其结构基于已表达(但未翻译)的人类？-防御素假基因的序列。广泛的、长期的目标：使用逆转录周期素来增强肺对病毒气雾剂的防御。明确的目标。1.检测逆周期蛋白对潜在的病毒生物恐怖因子的作用，包括甲型和乙型流感病毒(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)、痘苗病毒(痘病毒科)、VEE、甲型病毒(Togaviridae)和塔卡里布病毒(Arenaviridae)2.研究逆转录周期蛋白与表面活性蛋白SP-A和SP-D的相互作用3.检测RC2对肺表面活性物质磷脂单层和双层的作用。研究设计。逆转录周期蛋白-2(RC-2)及其类似物将通过固相多肽合成来制备。病毒将通过标准的空斑减少和CPE减少试验进行体外测试。逆周期蛋白对表面活性磷脂膜完整性的影响将通过使用Langmuir-Blodgett槽在单层体系中进行测试，并在双层体系中通过使用由表面活性脂类和相关的表面活性蛋白SP-B和SP-C组成的囊泡来测试。逆周期蛋白与SP-A和SP-D的结合将通过表面等离子体共振进行研究。在标准的空斑减少和CPE减少试验中，将使用从屠宰场获得的猪肺中纯化的蛋白质来研究逆周期蛋白对SP-A和SP-D(反之亦然)内在抗病毒活性的影响。与健康相关和机构使命。此应用程序直接响应PA 04-119。Layer语言摘要。许多自然病毒感染始于暴露在含有病毒的空气中。病毒气雾剂还可能允许生物恐怖分子传播使人丧失能力的病毒。我们建议测试逆周期蛋白(新发现的抗病毒多肽)对潜在的病毒生物恐怖主义因子的活性，并确定它们与ELF-覆盖在肺气囊表面的薄液体层-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.