Abstract
The risk of sepsis through bacterial transmission is one of the most serious problems in platelet transfusion. In processing platelet concentrates (PCs), several methods have been put into practice to minimize the risk of bacterial transmission, such as stringent monitoring by cultivation assays and inactivation treatment by photoirradiation with or without chemical agents. As another potential option, we applied a light-emitting diode (LED) with a peak emission wavelength of 265 nm, which has been shown to be effective for water, to disinfect PCs. In a bench-scale UV-LED exposure setup, a 10-min irradiation, corresponding to an average fluence of 9.2 mJ/cm2, resulted in >2.0 log, 1.0 log, and 0.6 log inactivation (mean, n = 6) of Escherichia coli, Staphylococcus aureus, and Bacillus cereus, respectively, in non-diluted plasma PCs. After a 30-min exposure, platelet counts decreased slightly (18 ± 7%: mean ± SD, n = 7); however, platelet surface expressions of CD42b, CD61, CD62P, and PAC-1 binding did not change significantly (P>0.005), and agonist-induced aggregation and adhesion/aggregation under flow conditions were well maintained. Our findings indicated that the 265 nm UV-LED has high potential as a novel disinfection method to ensure the microbial safety of platelet transfusion.
Highlights
Platelets play a central role in stopping bleeding under either low or high flow rate conditions through surface molecules including glycoproteins, such as GPIb/IX/V (CD42) and GPIIb/IIIa (CD41/CD61), and released molecules in platelet granules, which activate the surrounding platelets
The materials purchased for this study include: (1) phycoerythrin (PE)-conjugated anti-human CD62P, PE-conjugated mouse IgG, and peridinin chlorophyll protein complex (PerCP)-conjugated anti-human CD61 (BD Biosciences, Tokyo); (2) PE-conjugated anti-human CD42b antibody; (3) thrombin receptor-activating peptide (TRAP); (4) adenosine 5’-diphosphate (ADP) (Sigma Aldrich Japan, Tokyo); (5) collagen reagent (Horm collagen; Moriya Sangyo, Tokyo); (6) Escherichia coli (DH-5α; F, F80dlacZΔM15, Δ(lacZYA-argF) U169, deoR, recA1, endA1, hsdR17, phoA, supE44, λ, thi-1, gyrA96, relA1) (Takara Bio, Shiga, Japan); and (7) Staphylococcus aureus (NBRC 3060) and Bacillus cereus (NBRC 3001), gifted from the NITE Biological Resource Centre (Tokyo)
In control experiments without UV-light-emitting diode (LED) irradiation of the three bacterial species (E. coli, S. aureus, and B. cereus), a 100-μL aliquot taken from the spiked platelet concentrates (PCs) mixture at each time interval formed a total of 10–80 colonies on agar plates
Summary
Platelets play a central role in stopping bleeding under either low or high flow rate conditions through surface molecules including glycoproteins, such as GPIb/IX/V (CD42) and GPIIb/IIIa (CD41/CD61), and released molecules in platelet granules, which activate the surrounding platelets. PCs are usually stored at room temperature (20–24 ̊C) with continuous agitation; this storage condition allows any contaminating bacteria to proliferate quickly. Cold-stored platelets have markedly reduced in vivo survival after transfusion, despite enhanced platelet aggregability due to their activation under low temperatures, and room temperature storage consistently shows longer in vivo platelet circulation times [6, 7]. The shelf-life of PC products is relatively short (3 to 5 days) in countries without a bacterial screening system or pathogen inactivation system [8]. To mitigate the risk of bacterial contamination in PCs, two major strategies have been introduced in many advanced countries during the past two decades: culturebased detection of bacteria and pathogen inactivation procedures [8, 10, 11]
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