Use of 8-methoxypsoralen and long wavelength ultraviolet radiation for decontamination of platelet concentrates.
Use of 8-methoxypsoralen and long wavelength ultraviolet radiation for decontamination of platelet concentrates.
复制标题
使用 8-甲氧基补骨脂素和长波紫外线辐射对浓缩血小板进行净化。
DOI：
10.1117/12.137506
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Wiesehahn,G
中科院分区：
文献类型：
--
作者：
Corash,L;Lin,L;Wiesehahn,G
通过血液制品传播病毒性疾病仍然是输血医学中的一个问题。已经开发了多种方法来灭活血浆和血浆组分中的病毒病原体，包括：干加热、湿加热、溶剂-去污剂处理和免疫亲和纯化。虽然其中一些方法成功地灭活了病原病毒，但灭活可能不完全或导致不稳定的血浆蛋白和细胞受损。我们开发了一种用于浓缩血小板 (PC) 的光化学净化系统 (PCD)，利用长波紫外线辐射 (UVA，320 - 400 nm) 和 8-甲氧基补骨脂素 (8-MOP) 进行处理。该系统能够以 25 - 30 log/hr 的速度灭活 PC 中的细菌大肠杆菌或金黄色葡萄球菌、6 log/hr 的噬菌体 fd、0.9 log/hr 的噬菌体 R17 和 1.1 log/hr 的猫白血病病毒 (FeLV)。 PCD治疗6小时后，PCD治疗和对照PC的血小板完整性和功能相当。过夜储存后，PCD 处理的 PCD 和对照 PC 血小板特性相同，但与对照相比，PCD 处理的 PC 的 TXB-2 产量略有减少。 PCD处理后，PC储存48至96小时。 PCD处理和对照PC的血小板计数、形态评分、细胞外LDH水平、聚集反应、致密体(db)含量和α颗粒((α)g)含量具有可比性。我们评估了 PCD 技术灭活细胞内和细胞外病毒的能力，量化了污染淋巴细胞中 DNA 加合物形成的程度，并测量了聚合酶链式反应 (PCR) 介导的细胞内 DNA 扩增的抑制作用。添加到人血小板浓缩物（最终浓度 106）中的高滴度无细胞鼠巨细胞病毒在 30 分钟内被 PCD 灭活。将高水平感染猫鼻气管炎病毒 (FeRTV) 的猫肾成纤维细胞接种到 PC 中，然后进行 PCD 处理，在 10 分钟内灭活 4.8 个对数的 FeRTV。将纯化的人淋巴细胞接种到PC中并在3 H 8-MOP存在下用PCD处理。六小时的 PCD 处理导致每 1000 个碱基对 (bp) DNA 形成 9.3 至 12.8 个 8-MOP 加合物。检查了 HLA-DQ(α) 基因座 242 bp 片段的 PCR 扩增。 PCR DNA 扩增的抑制取决于形成的 8-MOP 加合物的数量，当每 1000 bp 形成超过 12 个加合物时，不存在扩增。这些研究表明，PCD 可以有效灭活接种到标准人 PC 中的高滴度细胞相关病毒和无细胞病毒。 DNA 加合物形成的效率可以定量，并且污染 PC 的淋巴细胞中 8-MOP 加合物形成的水平与报道的不存在血小板的细胞 DNA 中加合物形成的水平相当。
Transmission of viral diseases through blood products remains a problem in transfusion medicine. A number of methods have been developed to inactivate viral pathogens in plasma and plasma fractions, including: dry heating, wet heating, solvent-detergent treatment, and immunoaffinity purification. While some of these methods successfully inactivate pathogenic viruses, inactivation may be incomplete or result in damage to labile plasma proteins and cells. We have developed a photochemical decontamination system (PCD) for platelet concentrates (PC) utilizing treatment with long wavelength ultraviolet radiation (UVA, 320 - 400 nm) and 8-methoxypsoralen (8-MOP). This system is capable of inactivating 25 - 30 logs/hr of bacteria E. coli or S. aureus, 6 logs/hr of bacteriophage fd, 0.9 log/hr of bacteriophage R17 and 1.1 logs/hr of feline leukemia virus (FeLV) in PC. Immediately following 6 hrs of PCD treatment, platelet integrity and function of PCD treated and control PC were equivalent. After overnight storage PCD treated and control PC platelet properties were equal, but there was a slight reduction in TXB-2 production of PCD treated PC compared to controls. Following PCD treatment, PC were stored for 48 to 96 hrs. Platelet counts, morphology scores, extracellular LDH levels, aggregation response, dense body (db) content, and alpha granule ((alpha) g) content of PCD treated and control PC were comparable. We assessed the ability of the PCD technique to inactivate intracellular and extracellular virus, quantified the degree of DNA adduct formation in contaminating lymphocytes, and measured the inhibition of polymerase chain reaction (PCR) mediated amplification of intracellular DNA. High titers of cell-free murine cytomegalovirus added to human platelet concentrates (final concentration 106) were inactivated by PCD within 30 min. Cat renal fibroblasts infected at high levels with feline rhinotracheitis virus (FeRTV) were seeded into PC followed by PCD treatment with inactivation of 4.8 logs of FeRTV within 10 minutes. Purified human lymphocytes were seeded into PC and treated with PCD in the presence of3H 8-MOP. Six hours of PCD treatment resulted in the formation of 9.3 to 12.8 8-MOP adducts per 1000 base pairs (bp) of DNA. PCR amplification of a 242 bp segment at the HLA-DQ(alpha) locus was examined. Inhibition of PCR DNA amplification was dependent on the numbers of 8-MOP adducts formed, and no amplification was present when greater than 12 adducts per 1000 bp were formed. These studies indicate that PCD can effectively inactivate high titers of cell-associated and cell-free virus seeded into standard human PC. The efficiency of DNA adduct formation can be quantitated, and the level of 8-MOP adduct formation in lymphocytes contaminating PC is comparable to the level of adduct formation in cellular DNA reported in the absence of platelets.