【摘要】 目的探讨用核黄素光化学法预防输血相关移植物抗宿主病(TA-GVHD)的可行性。方法随机从的全血中收集淋巴细胞,将淋巴细胞悬浮于核黄素终浓度为50μmol/L的1640培养液中,注入440 nm处透光率为80%的PVC透光塑料袋内。用400—500 nm的可见光照射,照射量为8.8 J/ml。对照组为相同来源、悬浮于不含核黄素的1640培养液中,未接受光照处理的淋巴细胞。以植物血凝素(PHA)、CD3单抗、CD28单抗等同步刺激实验组和对照组淋巴细胞,用酶标方法检测细胞因子分泌量。结果经过核黄素光化学法处理后的淋巴细胞,接受PHA刺激后,实验组淋巴细胞IL-1β、IL-2、IL-4、IL-6、IL-8、IL-10、TNF-α和INF-γ的分泌量比对照组细胞分别降低了(99.47±0.80)%、(99.81±0.46)%、(84.26±24.20)%、(99.86±1.23)%、(92.30±11.04)%、(89.50±15.66)%、(99.98±0.06)%和100%;接受CD3单抗刺激后,实验组淋巴细胞IL-1β、IL-6、IL-8、IL-10、TNF-α和INF-γ的分泌量比对照组细胞分别降低了(99.32±1.30)%、(99.37±1.16)%、(93.31±7.86)%、(93.04±14.16)%、(84.70±10.22)%和100%。接受CD3单抗和CD28单抗联合刺激后,实验组淋巴细胞IL-1β、IL-6、IL-8、TNF-α和INF-γ的分泌量比对照组细胞分别降低了(96.88±7.25)%、(98.55±3.61)%、(94.35±4.93)%、(88.69±10.42)%和100%。结论核黄素光化学法处理可使淋巴细胞失去生成GVHD相关细胞因子的能力,初步证明核黄素光化学法能在细胞水平有效阻断淋巴细胞通过接受抗原诱导表达细胞因子参与TA-GVHD病理改变的通路。更多还原

【Abstract】 Objective To measure the effect of riboflavin photochemical treatment(RPT) on lymphocyte cytokine production in vitro and to discuse the feasibility of preventing transfusion-associated graft versus host disease(TA-GVHD) via RPT.Methods Lymphocytes,collected from random blood donors,and riboflavin,with a final concentration of 50 μmol/L,were added together into PVC transparent bags and then exposed to visible light,With the wave length ranged from 400 nm to 500 nm,under a final dose of 8.8 J/ml.Control lymphocytes were exposed to light in the absence of riboflavin.Lymphocytes exposed to riboflavin photochemical treatment(RPT-lymphocytes) and control lymphocytes were tested for the production of several cytokines in response to PHA and anti-CD3+anti-CD28.Results When RPT-lymphocytes were stimulated by PHA,the production of IL-1β,IL-2,IL-4,IL-6,IL-8,IL-10,TNF-α,and INF-γ were inhibited by(99.47±0.80)%,(99.81±0.46)%,(84.26±24.20)%,(99.86±1.23)%,(92.30±11.04)%,(89.50±15.66)%,(99.98±0.06)% and 100%,respectively.In anti-CD3 stimulated RPT-lymphocytes,the production of IL-1β,IL-6,IL-8,IL-10,TNF-α and INF-γ were inhibited by(99.32±1.30)%,(99.37±1.16)%,(93.31±7.86)%,(93.04±14.16)%,(84.70±10.22)% and 100%,respectively.When RPT-lymphocytes were stimulated by anti-CD3+anti-CD28,the production of IL-1β,IL-6,IL-8,TNF-α and INF-γ were inhibited by(96.88±7.25)%,(98.55±3.61)%,(94.35±493)%,(88.69±10.42)% and 100%,respectively,by RPT.The levels of cytokines present in the supernatant of RPT-lymphocytes after stimulation by antigens were significantly lower than those present in the corresponding supernatant of non-treated lymphocytes.Conclusion Riboflavin photochemical treatment with visible light inactivates lymphocytes and inhibits the production of cytokines.It appears to be a promising method to prevent immune reactions such as TA-GVHD.更多还原