PHOTOCHEMICAL DECONTAMINATION OF PLATELET CONCENTRATES

目前估计有500-700万的治疗性血小板输注 每年单位，随着器官使用的扩大，预计还会增加 移植和强化癌症化疗。健康风险 与血小板输注相关的主要原因是 血源性病毒感染剂传播的可能性 尽管进行了输血前的血液产品检测。这样做的目的是 建议是开发一种技术来灭活乙肝病毒(乙肝病毒)， 人类感染的非甲、非乙型肝炎病毒和巨细胞病毒 具有保存血小板功能的浓缩血小板(PC)。 来自该项目的信息将普遍适用于未来 对血液制品中其他病毒灭活的研究，以 这些过程对细胞和血浆成分的影响，以及 最终达到预防输血相关病毒疾病的目的。 开发了一种光化学去污(PCD)技术 利用8-甲氧基补骨脂素(8-MOP)和长波紫外线 辐照(UVA)灭活PC中的广谱病毒 体外保存血小板功能。这一项目旨在 优化PCD技术灭活细菌的效率 PC中细胞外和细胞内病毒的体外检测 为了量化病毒灭活的效率，开发动物 研究病毒灭活的模型，以证明其有效性 PCD在体内外对PC去污作用的研究 检测，并测定经PCD处理的血小板在体内的存活率 在灵长类动物中。[1]。黄曲霉毒素的光化学失活动力学 细胞外和细胞内人CN4V和小鼠CMV在PC中的应用 体外细胞培养系统和小鼠传染性模型将是 量过了。胞外和胞外的光化学灭活动力学 作为人类乙肝病毒模型的细胞内鸭肝炎病毒(DHBV)将被 在PC中用鸭肝细胞和鸡蛋感染性模型测定。 猫鼻气管炎病毒(FeRTV)将被用作 细胞内病毒，因为高水平的传染性可以在 带有此代理的PC。红细胞、淋巴细胞和血浆的作用 脂类污染对PC中病毒灭活的影响将被确定。在……里面 除8-MOP外，其他补骨脂素衍生物[4，5‘，8-三甲基补骨脂素 (TMP)和4‘-氨甲基-4，5’，8-三甲基补骨脂素(AMT)] 对PCD工艺进行了优化研究。[2]。细胞内化程度 将测量光加合物的形成以评估 前病毒DNA/RNA交联和聚合酶链式反应 将使用扩增试验来量化抑制效率。 核转录和病毒DNA转录。[3]。在确定...之后 基于病毒灭活研究的最佳灭活条件 这些条件对PCD处理的PC Will体外功能的影响 被测量。血小板功能将通过测量进行评估：血小板 产量、胞外pH、形态评分、聚集反应、 分泌、细胞外乳酸脱氢酶、血栓素B-2 产量、致密体含量和颗粒物含量。[4]。个人电脑，转到 添加了哪些已知数量的乙肝病毒和NANBV，并在 在各种PCD条件下，将被注入黑猩猩体内 对肝炎发病时间的评估。[5]。体内恢复， 经PCD治疗的食蟹猴的存活和止血功能 在正常和血小板减少的食蟹猴身上进行测量。

Therapeutic platelet transfusion is currently estimated at 5-7 million units annually, and is expected to increase with expanded use of organ transplantation and intensive cancer chemotherapy. The health risks associated with platelet transfusion are significant primarily because of the potential for transmission of blood-home viral infectious agents despite pretransfusion blood product testing. The objective of this proposal is to develop a technique to inactivate hepatitis B virus(HBV), hepatitis non-A, non-B virus(NANBV), and cytomegalovirus(CMV) in human platelet concentrates(PC) with preservation of platelet function. Information from this project will be generally applicable to future studies of the inactivation of other viruses in blood products, to the effects of these processes on cellular and plasma components, and ultimately to the prevention of transfusion-associated viral disease. A photochemical decontamination(PCD) technique has been developed utilizing 8-methoxypsoralen (8-MOP) and long wavelength ultraviolet irradiation (UVA) to inactivate a wide spectrum of viruses in PC with preservation of in vitro platelet function. This project seeks to optimize the efficiency of the PCD technique for inactivation of extracellular and intracellular viruses in PC, to develop in vitro assays to quantify the efficiency of viral inactivation, to develop animal models for study of viral inactivation, to demonstrate the effectiveness of the PCD treatment for decontamination of PC using in vitro and in vivo assays, and to determine the in vivo survival of PCD treated platelets in primates. [1]. The photochemical inactivation kinetics of extracellular and intracellular human CN4V and murine CMV in PC using in vitro cell culture systems and a murine infectivity model will be measured. The photochemical inactivation kinetics of extracellular and intracellular duck hepadna virus(DHBV), a model for human HBV, will be determined in PC using duck hepatocyte and egg infectivity models. Feline rhinotracheitis virus(FeRTV) will be used as a model of intracellular virus, since high levels of infectivity can be achieved in PC with this agent. The effects of red cell, lymphocyte, and plasma lipid contamination on viral inactivation in PC will be determined. In addition to 8-MOP, other psoralen derivatives [4, 5', 8-trimethylpsoralen (TMP) and 4'-aminomethyl-4, 5', 8-trimethylpsoralen (AMT)] will be studied to optimize the PCD technique. [2]. The degree of intracellular photoadduct formation will be measured to assess the efficiency of proviral DNA/RNA crosslinking, and the polymerase chain reaction (PCR) amplification assay will be used to quantify the efficiency of inhibition of nuclear and viral DNA transcription. [3]. After determination of optimal inactivation conditions based on viral inactivation studies, the effects of these conditions on in vitro function of PCD treated PC will be measured. Platelet function will be assessed by measuring: platelet yield, extracellular pH, morphology score, aggregation response, secretion, extracellular lactate dehydrogenase, thromboxane B-2 production, dense body content and granule content. [4]. Human PC, to which known amounts of HBV and NANBV have been added, treated under various PCD conditions, will be transfused into chimpanzees followed by evaluation for the onset of hepatitis. [5]. The in vivo recovery, survival and hemostatic function of PCD treated cynomolgus PC will be measured in normal and thrombocytopenic cynomolgus monkeys.