IN VITRO INACTIVATION OF VIRUSES IN BLOOD COMPONENTS

血液成分中病毒的体外灭活

• 批准号: 2519450
• 负责人: GIRISH N VYAS
• 金额: $ 41.28万
• 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/2519450
• 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.

传播细胞相关人类免疫缺陷病毒的风险 (HIV-1/-2)、人类 T 细胞嗜淋巴细胞病毒 (HTLV-I/-II) 和 巨细胞病毒 (CMV)，以及传播无细胞 HIV 和肝炎 经过全面筛查的血液中的 B 型和 C 型病毒（HBV 和 HCV）可以减少 通过血源性病毒（BBV）的体外灭活/去除（IVIR）。我们 提出了 BBV IVIR 整体实施的三步级联流程 血液常规处理时的血液：（1）选择性去除白细胞 血细胞，含有并允许细胞相关 BBV 的复制； (2) 受体介导从无白细胞中去除无细胞 BBV 全血； (3)残留BBV的生化或化学灭活 通过添加与生物相容的药理学上可接受的试剂 血。为了测试这个概念，我们的实验设计采用 HIV 作为 BBV 原型，因为 HIV 既存在于细胞相关的细胞中，也存在于细胞外的细胞中 形成并可以通过补充体外定量监测 通过聚合酶链式反应 (PCR) 进行生化扩增，以及 通过与 CD4 阳性细胞共培养进行生物学分析。因为 在整个白细胞耗尽期间发生不可接受的血小板损失 血液，目前使用两种不同的聚酯过滤器来包装红色 细胞和血小板浓缩物。添加生物素化受体 蛋白质 (CD4-CD26) 或 HIV 包膜蛋白的人类抗体，结合 链霉亲和素包被的磁珠，将能够特异性捕获 来自白细胞过滤的无细胞 HIV 和 HIV 免疫复合物 血。我们将研究用于酶促作用的多靶点核酶 HIV RNA 灭活和/或抗坏血酸化学灭活 残留病毒粒子。 HIV 研究结果将提供 无细胞 HBV 和 HCV 的类似 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 红细胞和血小板，我们将进行血液学检查 验证 IVIR 血液治疗的安全性和有效性 BBV。最终，我们的研究和开发工作可能会达到顶峰 与能够提供集成的制造商的合作 系统适用于输血实践中 BBV 的常规 IVIR。