IN VITRO INACTIVATION OF VIRUSES IN BLOOD COMPONENTS

血液成分中病毒的体外灭活

• 批准号: 2231266
• 负责人: GIRISH N VYAS
• 金额: $ 38.03万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-30 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2231266
• 关键词: CD4 molecule; biotin; blood /lymphatic pharmacology; blood filtration; blood transfusion; blood treatment; disease /disorder prevention /control; disease /disorder proneness /risk; enzyme activity; human immunodeficiency virus; immune complex; leukocytes; polymerase chain reaction; polymers; ribozymes; virion; virus envelope

The risk of transmitting cell-associated human immunodeficiency viruses (HIV- 1/-2), human T cell lymphotropic viruses (HTLV-I/-II), and cytomegaloviruses (CMV), and of transmitting cell-free HIV and hepatitis B and C viruses (HBV and HCV) from fully-screened blood can be reduced by in vitro inactivation/removal (IVIR) of blood-borne viruses (BBV). We propose a three-step cascade process for IVIR of BBV carried out on whole blood during routine blood processing: (1) selective removal of white blood cells, which harbor and permit replication of cell-associated BBV; (2) receptor-mediated removal of the cell-free BBV from leukocyte-free whole blood; and (3) biochemical or chemical inactivation of residual BBV by adding pharmacologically-acceptable agents that are biocompatible with blood. For testing this concept, our experimental design employs HIV as a prototype BBV, because HIV occurs both in cell-associated and cell-free forms and can be monitored quantitatively by complementary in vitro amplifications, biochemically by the polymerase chain reaction (PCR) and biologically by co-cultivation with CD4-positive cells. Because unacceptable platelet loss occurs during leukocyte depletion of whole blood, two different polyester filters are currently used for packed red cells and platelet concentrates. The addition of biotinylated receptor proteins (cD4-CD26) or human antibodies to HIV envelope proteins, bound to streptavidin-coated magnetic beads, will enable specific capture of both cell-free HIV and HIV-immune-complexes from the leukocyte-filtered blood. We will investigate multi-target ribozymes for enzymatic inactivation of HIV RNA and/or ascorbic acid for chemical inactivation of residual virions. Results of the studies with HIV will provide an applicable paradigm for analogous IVIR of cell-free HBV and HCV. Although we do not expect significant changes in membrane structure and function of red cells and platelets, we will perform hematological investigations to validate the safety and effectiveness of blood treatment for IVIR of BBV. Ultimately, our research and development efforts could culminate in a collaboration with manufacturer(s) capable of providing an integrated system suitable for routine IVIR of BBV in transfusion practice.

传播细胞相关的人类免疫缺陷病毒的风险 (HIV-1/-2)，人类T细胞嗜淋巴病毒(HTLV-I/-II)，以及 巨细胞病毒(CMV)以及传播无细胞艾滋病毒和肝炎的病毒 可以减少来自全面筛查血液的B和C病毒(乙肝病毒和丙型肝炎病毒) 通过体外灭活/去除(IVIR)血液传播病毒(BBV)。我们 提出了BBV整体IVIR的三步级联流程 常规血液加工过程中的血液：(1)选择性去除白色 血细胞，容纳并允许复制与细胞相关的BBV； (2)受体介导的无细胞BBV从无白细胞中去除 全血；(3)残留BBV生化或化学灭活 通过添加生物相容的药理上可接受的药物 血。为了测试这一概念，我们的实验设计使用HIV作为 BBV原型，因为HIV既发生在细胞相关的，也发生在无细胞的情况下 形成并可通过体外互补进行定量监测 通过聚合酶链式反应(PCR)生化扩增和 通过与CD4阳性细胞共培养从生物学上讲。因为 在全血白细胞耗尽过程中发生不可接受的血小板丢失 血液，目前有两种不同的聚酯过滤器用于包装红色 细胞和血小板浓缩物。生物素化受体的添加 结合的HIV包膜蛋白(CD4-CD26)或人抗体 链霉亲和素包被的磁珠，将使特定捕获 白细胞过滤后的无细胞HIV和HIV免疫复合体 血。我们将研究酶的多靶点核酶 HIV RNA和/或抗坏血酸的化学灭活 残留的病毒粒子。对艾滋病毒的研究结果将提供一种 无细胞乙肝病毒和丙型肝炎病毒类似IVIR的适用范例。虽然 我们预计膜结构和功能不会发生重大变化 对于红细胞和血小板，我们将进行血液学检查 验证静脉内胰岛素抵抗血液治疗的安全性和有效性 BBV。最终，我们的研究和开发努力可能会达到 与制造商(S)合作，能够提供完整的 该系统适用于输液实践中BBV的常规IVIR。