Wagner S J, Storry J R, Mallory D A, Stromberg R R, Benade L E, Friedman L I
Product Development Laboratory, American Red Cross Blood Services, Rockville, Maryland.
Transfusion. 1993 Jan;33(1):30-6. doi: 10.1046/j.1537-2995.1993.33193142306.x.
Several red cell storage properties were evaluated following phototreatment with methylene blue (MB) under conditions that inactivated > or = 6 log10 of added vesicular stomatitis virus. Red cell 2,3 DPG levels were similar to untreated controls throughout conventional 42-day storage at 4 degrees C. Plasma hemoglobin levels were elevated approximately twofold in MB-phototreated samples, and morphology scores were 5 percent lower after 42-day storage. ATP levels declined 30 percent in phototreated samples and in a control sample containing MB and not exposed to light. Lipid peroxidation was not observed in treated or control cells, nor were differences observed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis of ghost membranes derived from phototreated and control samples. Phototreated cells exhibited enhanced ion permeability; sodium and potassium levels approached equilibrium with the suspending medium within 4 to 7 days after treatment. Direct agglutination tests using rabbit anti-human IgG or rabbit anti-human serum albumin on MB-phototreated cells indicated that serum proteins had absorbed to the surface of treated red cells. Plasma depletion by washing red cells prior to phototreatment did not prevent protein binding upon subsequent addition of untreated autologous or group AB plasma. To a much smaller extent, phototreatment of plasma resulted in IgG association with untreated red cells. The addition of glutathione to red cell suspensions prevented IgG binding to phototreated red cells but did not prevent enhanced ion permeability. Taken together, these data suggest that the red cell surface is altered by virucidal MB phototreatment of vesicular stomatitis virus.
在能使添加的水泡性口炎病毒失活≥6 log10的条件下,用亚甲蓝(MB)进行光处理后,评估了几种红细胞储存特性。在4℃常规储存42天的整个过程中,红细胞2,3-二磷酸甘油酸(2,3-DPG)水平与未处理的对照相似。MB光处理样本中的血浆血红蛋白水平升高了约两倍,42天储存后形态学评分降低了5%。光处理样本和含有MB但未暴露于光的对照样本中,三磷酸腺苷(ATP)水平下降了30%。在处理的或对照的细胞中均未观察到脂质过氧化,来自光处理和对照样本的空壳细胞膜的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳也未观察到差异。光处理的细胞表现出离子通透性增强;处理后4至7天内,钠和钾水平与悬浮介质接近平衡。使用兔抗人免疫球蛋白(IgG)或兔抗人血清白蛋白对MB光处理细胞进行直接凝集试验表明,血清蛋白已吸附到处理过的红细胞表面。在光处理前洗涤红细胞以去除血浆,并不能阻止在随后添加未处理的自体或AB型血浆时蛋白质的结合。在小得多的程度上,血浆光处理导致IgG与未处理的红细胞结合。向红细胞悬液中添加谷胱甘肽可阻止IgG与光处理的红细胞结合,但不能阻止离子通透性增强。综上所述,这些数据表明,对水泡性口炎病毒进行杀病毒的MB光处理会改变红细胞表面。