IN VITRO INACTIVATION OF VIRUSES IN BLOOD COMPONENTS

血液成分中病毒的体外灭活

• 批准号: 2231258
• 负责人: DANIEL MERUELO
• 金额: $ 35.08万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2231258
• 关键词: Retroviridae; SDS polyacrylamide gel electrophoresis; antiviral agents; blood transfusion; chemical structure function; covalent bond; crosslink; cyclic compound; drug screening /evaluation; erythrocytes; polymerase chain reaction; virus RNA; virus envelope; virus protein; western blottings

DESCRIPTION (Adapted from the applicant's abstract) This application proposes studies to evaluate the potential for development of hypericin (HY) and of numerous structurally related aromatic polycyclic diones, as virus inactivators in red cell concentrates for transfusion. The studies are based on encouraging preliminary data which show lack of adverse effects to red blood cells (RBC) at effective virucidal doses. They aim to define the optimal conditions for obtaining complete inactivation of very high titers of cell free virus in a blood environment. Hypericin is an effective virucidal agent which directly inactivates a broad range of enveloped viruses. Hypericin is a photodynamic molecule, which is also active in vivo, presumably due to a low red/ox potential. Thus, HY is currently being evaluated in clinical trials in AIDS patients, and phase I of the trials confirmed the transfusibility of HY. The problem of dual transmission of HIV infection via cell-free virus and through latently infected PBMC will be addressed by evaluating combinations of HY and leukocyte filtration in achieving complete sterility, and the possibility of inflicting photodynamic lesions to infected PBMC. The application will address aspects of the mechanism of the virucidal action of HY. The modifications in viral proteins HIV p55gag, p24 and RT, which are targeted for photodynamic cross-linking by HY will be characterized, and other affected virus proteins identified. The potential involvement of viral RNA in RNA- protein covalent complexes will be evaluated. The investigators propose clarify whether the action of HY covalently cross links viral genomic RNA, with RT and other core proteins. Amplification of RNA from HY treated virions using RNA PCR will be utilized to identify genomic regions cross linked by HY. The question whether HY itself or only photodynamically generated excited oxygen species affect the capsids will be examined by isolation of intact HIV cores and examination of their susceptibility to HY and light. These studies have implications for potential inactivation of non enveloped viruses (such as B19 parvovirus) in blood by HY. RT activity and capsid protein migration patterns on SDS-PAGE and Western blots will be used as assays. Structural HY analogs with methyl group substitutions and alterations in the polycyclic aromatic skeleton will be evaluated for potentially improved efficacies and for reduced binding to RBC. The virucidal activities of HY ion pairs with cationic metals, basic amines and amino acids, and their interactions with physiological transport proteins, will be studied in comparison with the standard HY-Na+. Potential for adverse effects to blood components including clinically significant parameters of red cell surface membrane integrity, (2,3-DPG, hemolysis, normal expression of ABO, Rh, minor blood group antigens and possible deposition of IgG or complement on RBC), by treatment with HY will be examined. Potential methods for removal of HY from blood after virus inactivation, incorporating hydrophobic resins and other methods will be evaluated. The applicant organization will collaborate with the following sites: 1) New York Blood Center, 2) Oklahoma Blood Institution, and 3) Weizmann Institute of Science.

描述（根据申请人的摘要改编）此申请 提出的研究以评估金丝桃素的发展潜力 （hy）和许多与结构相关的芳族芳族多环狄酮的 作为红细胞浓缩物中输血的病毒灭活剂。这 研究基于令人鼓舞的初步数据，这些数据表明缺乏 在有效的病毒剂量下对红细胞（RBC）的不利影响。 他们旨在定义获得完整的最佳条件 血液中非常高的无细胞病毒滴度灭活 环境。金丝桃素是一种有效的病毒剂，直接 使广泛的包膜病毒失活。 Hyperinin是A。 光动力分子在体内也很活跃，大概是由于 低红/牛电位。因此，目前正在评估HY AIDS患者的临床试验，该试验的第一阶段证实 Hy的吸收性。 HIV感染双重传播的问题 通过无细胞病毒和通过潜在感染的PBMC解决 通过评估HY和白细胞过滤的组合 完全不育，并可能导致光动力 感染PBMC的病变。该应用程序将解决 Hy病毒作用的机制。病毒的修饰 蛋白质HIV P55GAG，P24和RT，针对光动力学 通过hy的交联，将表征其他受影响的病毒 蛋白质确定。病毒RNA参与RNA- 将评估蛋白质共价复合物。调查人员建议 澄清Hy的作用是否共价交叉链接病毒基因组 RNA，带有RT和其他核心蛋白。从HY扩增RNA 使用RNA PCR处理的病毒体将用于鉴定基因组 由hy连接的区域交叉。问题是hy本身还是 光动力生成的激发氧影响衣壳 将通过隔离完整的艾滋病毒核心检查并检查 他们对hy and Light的敏感性。这些研究有含义 潜在的非包膜病毒的失活（例如B19 细小病毒）在血液中。 RT活性和衣壳蛋白迁移 SDS-PAGE和Western印迹上的图案​​将用作测定。 与甲基取代的结构类似物和变化 将评估多环芳族骨骼的潜在 提高效力并减少与RBC的结合。病毒性 与阳离子金属，碱性胺和氨基配对的活动 酸及其与生理转运蛋白的相互作用， 将与标准HY-NA+相比进行研究。潜力 对血液成分的不利影响，包括临床意义 红细胞表面膜完整性的参数（2,3-DPG，溶血， ABO，RH，次要血型抗原的正常表达和可能 IgG的沉积或在RBC上的补充），通过使用Hy处理将是 检查。病毒后从血液中取出HY的潜在方法 灭活，结合疏水树脂和其他方法将 进行评估。 申请人组织将与以下网站合作： 1）纽约血液中心，2）俄克拉荷马州的血液机构，3）Weizmann 科学研究所。