title: Use of 8-methoxypsoralen and long wavelength ultraviolet radiation for decontamination of platelet concentrates.
abstract: Transmission of viral diseases through blood products remains a problem in transfusion medicine. 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-methoxypsorlan (8-MOP). This system is capable of inactivating 25-30 logs/hour of bacteria E. coli or S. aureus, 6 logs/hour of bacteriophage fd, 0.9 log/hour of bacteriophage R17, and 1.1 logs/hour of feline leukemia virus (FeLV) in PC. Immediately following 6 hours 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 hours. 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 10(6)) were inactivated by PCD within 30 minutes. 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 of 3H 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.
authors.affiliation: Department of Laboratory Medicine, University of California, San Francisco 94143.
authors.forename: L
authors.initials: L
authors.lastname: Corash
authors.forename: L
authors.initials: L
authors.lastname: Lin
authors.forename: G
authors.initials: G
authors.lastname: Wiesehahn
mesh_terms: D016913:Blood Component Transfusion / Q000009:adverse effects / Q000592:standards*; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology* / Q000528:radiation effects / Q000648:ultrastructure; D001793:Blood Preservation; D002478:Cells, Cultured; D004249:DNA Damage; D004261:DNA Replication / Q000187:drug effects / Q000528:radiation effects; D005347:Fibroblasts / Q000187:drug effects / Q000382:microbiology / Q000528:radiation effects; D006526:Hepatitis C / Q000453:epidemiology / Q000517:prevention & control / Q000635:transmission; D006801:Humans; D015994:Incidence; D007962:Leukocytes / Q000187:drug effects / Q000528:radiation effects; D008730:Methoxsalen / Q000494:pharmacology* / Q000528:radiation effects; D010777:Photochemistry; D010974:Platelet Aggregation / Q000187:drug effects / Q000528:radiation effects; D013929:Thromboxane B2 / Q000096:biosynthesis; D014466:Ultraviolet Rays; D014777:Virus Diseases / Q000517:prevention & control* / Q000635:transmission; D018406:Virus Physiological Phenomena; D014779:Virus Replication / Q000187:drug effects / Q000528:radiation effects; D014780:Viruses / Q000187:drug effects* / Q000528:radiation effects
chemical_list: D013929:Thromboxane B2; D008730:Methoxsalen