Experimental Studies On The Virus And Cytokines Inhibition In Platelet Suspension Via Ribolfavin Photochemical Treatment

Pathogen sterilization of blood and the inhibition of leukocyte derived cytokinesin blood component is an important clinical tool to ensure transfusion safety. Thetechnology of pathogen disinfection is relatively available in plasma and somemeasures have been applied clinically. However, no approach has been reported inthe red blood cells. Previous investigations showed that methylene blue photochemicaland psoralen derivatives S-59can effectively inactivate virus and leukocytes inplatelets, but the platelets treated with the MB photochemical were negativelyaffected in FⅧ and fibrinogen and additionally, MB is to some extent, toxic. Specialequipment is needed to remove the disinfectant for the blood treated by S-59.Therefor both methods have flaws for platelets sterilization. Riboflavin is anessential water-soluble vitamin which exists extensively in human organs, and afteractivation by UV/visible light leads to the fragmentation of nucleic acid throughelectron transfer and covalent binding to the guanine of the pathogen nucleic acid.The present work,used Riboflavin plus UV light treatment to sterilized selectedvirus in apheresis platelet suspension, and at the same time, inhibits the productionof cytokines by residual leukocytes during storage.Objective To explore the safety and efficacy of riboflavin UV-light joint inactivationof virus, and the inhibition of leukocytes derived cytokines in platelets preservation andtransfusionMethods In experimental group, human cytomegalovirus (HCMV AD169) was mixedwith the platelets suspension in final concentration of150μmol/L of riboflavin solution.The sample was put into the apheresis platelets, followed by UV-light sterilization, andthe viral titer was tested in platelet suspension. The production of cytokines were detected by ELISA on0d,3d, and5d after irradiation, and the change of plateletsparameter was observed. Negative control group was composed of fresh apheresisplatelets from the same collection without any treatments, cytokines weresynchronously detected. Two groups of platelets were subjected to phytohaemagglutinin(PHA) for synchronous stimulation.Results A dose of1500mJ/cm2UV light combined with riboflavin gave rise to aeffective sterilization of virus in the platelets suspension, however the slight increase ofthe expression of leukocyte-derived cytokines was noted after day3and day5post-treatment(P<0.05). The production of cytokines were dramatically enhanced afterPHA administration in negative control groups（P<0.05）.Conclusions Riboflavin plus UV light treatment could significantly syerilize the virus,inhibit the cytokines production of residual leukocytes of PLT during storage, and invitro apheresis platelets’parameters showed no significant difference compared withnegative control.