THIAZINE DYE MEDIATED PHOTOKILLING OF HIV 1 VIRUSES

描述（改编自申请人摘要） 研究人员发现，光敏染料亚甲基蓝（MB） 加光（MB+L）非常有效地灭活HIV-1病毒， 培养的人外周血单个核细胞。 MB非常低（约24 nM），所需的光也很低 短时间内的强度。 此外，MB具有良好的 用于血液成分的吸收光谱。 内容因此 对这种方法的改进可能有助于抑制HIV病毒 在血液成分中。 MB+L对HIV-1的抑制作用极有可能 传播是由单线态氧机制介导的， 致命的损伤发生在非常特定的目标上。相对于其他 可能的光敏方法，其中作用机制似乎 基于对MB+L的广泛机制研究， 在申请人的实验室中杀死RNA病毒，研究人员 我认为HIV-1病毒的MB+L致死靶点是RNA， 二级结构区，可能位于特定的必需RNA/蛋白质 交互界面 申请人希望进行基础研究 这将有助于我们了解MB+L介导的机制基础， 灭活艾滋病毒，并利用所获得的知识来设计和测试 这些策略将实现病毒灭活的改善， 对血液成分如血小板造成严重损害 浓缩物。 研究人员提出了一系列的实验 旨在： 了解MB+L对HIV传播的损伤部位和性质。 申请人计划确定是否通过以下方式降低HIV感染性： 用MB+L处理分离的HIV病毒体。 如果病毒体受损， 将确定病毒RNA是否受损，并确定 致命病变发生的特定区域（残留物），并确定 如果它的形成是由单线态氧介导的话。 还将 确定MB+L是否损伤病毒蛋白或病毒包膜脂质， 确定所涉及的机制以及 损害 还将确定细胞中是否存在HIV RNA （组装前）被MB+L损坏。 研究人员还将设计并测试一系列亚甲基蓝 基于（噻嗪）的衍生物，以努力提高特异性， 消除艾滋病毒感染风险的有效性。 最后，研究人员计划制定一个程序， 在上述基础研究中获得的知识将进一步 消除血液成分存在时感染艾滋病毒的风险 而不会严重损害血液成分。

DESCRIPTION (Adapted from the applicant's abstract) The investigators have found that the photoactive dye methylene blue (MB) plus light (MB+L) very effectively inactivates the HIV-1 virus in cultured human peripheral blood mononuclear cells.The effective dose of MB is very low (about 24 nM) and the light required is also of low intensity for a short duration. In addition, MB has a favorable absorption spectra for use with blood components. Therefore an adaptation of this approach may be useful to inactivate the HIV virus in blood components. It is highly likely that MB+L inhibition of HIV-1 propagation is mediated by a singlet oxygen mechanism and that the lethal lesion occurs at a very specific target. In contrast to other possible photoactive approaches where the mechanism of action appears to be at the viral membrane, based on extensive mechanistic studies on MB+L killing of RNA viruses in the applicant's laboratory, the investigators think that the MB+L lethal target of the HIV-1 virus is at a RNA secondary structure area, probably at a specific essential RNA/protein interaction interface. The applicants want to conduct basic studies which will help us understand the mechanistic basis to MB+L mediated inactivation of HIV and utilize the knowledge gained to design and test strategies that will achieve improvement in viral inactivation without causing significant damage to blood components, such as platelet concentrates. The investigators propose a series of experiments designed to: Understand the site and nature of MB+L damage to HIV propagation. Applicants plan to determine whether HIV infectivity is diminished by treating isolated HIV virions with MB+L. If the virions are damaged, it will be determined if the viral RNA is damaged and ascertain the specific region (residue) where the lethal lesion occurs and determine if its formation is mediated by singlet oxygen. It will also be determined if MB+L damages viral proteins or viral envelope lipids and ascertain the mechanisms involved as well as the specificity of the damage. It will also be determined if the HIV RNA present in cells (prior to assembly) is damaged by MB+L. The investigators will also design and test a series of methylene blue based (thiazine) derivatives in an effort to improve the specificity and efficacy in eliminating the risk of HIV infections. Finally the investigators plan to develop a procedure to apply the knowledge obtained in the above basic studies that will further eliminate the risk of HIV infection in the presence of blood components without significantly damaging the blood components.