IN VITRO INACTIVATION OF VIRUSES IN BLOOD COMPONENTS

传播细胞相关人类免疫缺陷病毒的风险 (HIV-1/-2）、人T细胞嗜淋巴细胞病毒（HTLV-I/-II），和 巨细胞病毒（CMV），以及传播无细胞艾滋病毒和肝炎 可减少来自全面筛查血液的B和C病毒（HBV和HCV） 通过体外灭活/清除（IVIR）血源性病毒（BBV）。我们 提出了一个整体进行的BBV IVIR的三步级联工艺 常规血液处理过程中的血液：（1）选择性去除白色 血细胞，其携带并允许细胞相关BBV的复制; (2)受体介导的无细胞BBV从无白细胞的 全血;和（3）残留BBV的生物化学或化学灭活 通过加入药理学上可接受的药剂， 血为了测试这一概念，我们的实验设计采用艾滋病毒作为 一个原型BBV，因为艾滋病毒发生在细胞相关和无细胞 形成，并可通过体外互补 扩增，通过聚合酶链式反应（PCR）进行生物化学扩增， 生物学上通过与CD 4阳性细胞共培养。因为 在全血白细胞耗竭期间发生不可接受的血小板损失。 血液，两种不同的聚酯过滤器目前用于包装红 细胞和血小板浓缩物。加入生物素化受体 蛋白（CD 4-CD 26）或HIV包膜蛋白的人抗体，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 血我们将研究多靶点核酶用于酶促反应， 用于化学灭活HIV RNA和/或抗坏血酸灭活 残留的病毒体对艾滋病毒的研究结果将提供一个 适用于无细胞HBV和HCV的类似IVIR的范例。虽然 我们并不期望膜结构和功能发生显著变化， 红细胞和血小板，我们将进行血液学研究 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高潮， 与能够提供集成的制造商合作 适用于输血实践中BBV常规IVIR系统。

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.