| Background Blood transfusion is an important means to save life in early modern medicine,on the other hand,blood transfusion is likely to exist the risk that blood recipient may encounter blood-transmitted pathogen infection,At present,the main method of preventing transfusion-transmitted diseases is blood-transmitted pathogens screening,but because of pathogens gene mutation,immunogenic changes and infected "window period",the small probability of undetected events is unavoidable in the conditions of using conventional detection techniques.Not to mention to the blood test coverage is very limited,apart from AIDS virus (HIV),hepatitis B virus (HBV), hepatitis C virus(HCV) and microspironema pallidum.at least dozens of blood-transmitted pathogens are not included in the detection range,including the human T-cell leukemia virus(HTLV),cytomegalo virus(MCV),EB virus(EBV),therefore,it is impossible to completely eliminate the risk of transfusion-transmitted diseases only rely on blood tests,and blood components pathogen inactivation technology is considered to be most reliable barrier to fundamentally eliminate transfusion-associated pathogens infection and ensure the safety of blood transfusion,thus,the study of blood pathogen inactivation methods and mechanisms become a hot research in transfusion medicine.Blood pathogen inactivation technologies include methylene blue photochemical method,psoralen S-59 method,riboflavin photochemical method,S-303 pathogen inactivation technology,PEN-110 pathogen inactivation technology and so on.Pathogen inactivation in plasma mainly use methylene blue photochemical method, psoralen S-59 method and riboflavin photochemical method are used for pathogen inactivation in platelet.Currently,there is not whole blood pathogen inactivation technology.Allogeneic lymphocytes will proliferate heavy in immunocompromised recipients body which will eventually consider it as foreign body,and form transfusion-associated graft-versus-host disease(TA-GVHD),the mortality rate of the disease up to 90%,therefore allogeneic lymphocytes are considered special pathogens in transfusion medicine.Now we have known riboflavin photochemical method can effectively inactive lymphocytes,but it lack indepth research about whether it have adverse effect on red blood cells.This study was designed to investigate the inactivation of lymphocytes and the effect of red blood cells treated by riboflavin photochemical method under the excitation of visible,providing methodological practice and data accumulation for the eventual establishment of riboflavin photochemical pathogen inactivation of red blood cells even whole blood.Objective We studied the effects on deformability,membrane structure,morphology,metabolic capacity,vitality and function of red blood cells,on lymphocyte cell morphology and activity, proliferation,apoptosis status and cytokine secretion,when they were treated by riboflavin photochemical method under the excitation of visible.Methods 1) The whole blood was divided into two groups:an experimental group and a control group. Riboflavin (final concentration:100μmol/L) was added into experimental group, putting the bag in a 4℃ pathogen inactivation tank, and 420nm visible light irradiated the bag, the irradiation dose was 40J/mL. The control group without riboflavin was not illuminate.2) The potassium levels, hemoglobin oxygen saturation, glucose content, lactic acid content of red blood cells in whole blood were detected by blood gas analyzer in Od,7 d,21d,35d,42d.3) Red blood cells were separated from whole blood and resuspended into a certain percentage preservation solution, red blood cells were detected erythrocyte osmotic fragility, free hemoglobin, hemolysis rate,red blood cell count(Red blood cell count,RBC), hematocrit (Hematocrit,Hct), hemoglobin content(Hemoglobin,Hgb), ATP content and 2,3-DPG content, saving Od,7d,21d,35d,42d.4) Lymphocytes were separated in vitro,and CD3/CD28 antibodies stimulated proliferation.Cell morphology,activity, on inactivation of lymphocytes treated by riboflavin photochemical before and after CD3/CD28 antibody stimulation.Results 1. Riboflavin photochemical methods had no effect on red blood cell osmotic fragility;on red blood cell morphology such as:red blood cell count, hemoglobin, hematocrit;on red blood cell metabolism:glucose and lactic acid content; on red blood cell activity such as ATP;on function:hemoglobin oxygen saturation,2,3-DPG; it had an impact on free hemoglobin and red blood cell K+ concentration, but the rate of hemolysis still meet US,EU and our country standards.2. Riboflavin photochemical method changed morphological, decreased activity,proliferation, enhanced lymphocyte apoptosis and cytokine secretion.Conclusions Riboflavin photochemical method had an effect on the deformability,membrane structure,morphology, metabolic capacity, vitality and function of red blood cells within an acceptable range, and effect of inactivation of lymphocytes was good. |
PDF Full Text Request