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目的 研究甲氧基聚乙二醇 苯并三唑 (mPEG BTC)修饰淋巴细胞表面人类白细胞抗原 (HLA)对细胞有无损伤。方法 利用微量淋巴细胞毒性实验和单向混合淋巴细胞培养来检测mPEG BTC的修饰效果 ;利用电镜观察修饰前后淋巴细胞的形态 ;通过淋巴细胞转化实验及培养上清液的IL 2含量的检测对淋巴细胞的增殖、分化及分泌功能进行评价 ;检测淋巴细胞表面CD分子评价淋巴细胞的抗原识别功能 ;体外保存淋巴细胞检测其寿命 ;对淋巴细胞染色体进行分析 ,评价mPEG对细胞遗传物质的影响。结果修饰后淋巴细胞的微量淋巴细胞毒性实验结果 ,淋巴细胞转化率 ,淋巴细胞相对转化指数 ,IL 2分泌含量分别由修饰前的 (8.0± 0 )分 ,(6 3.6± 7.8) % ,(1.0 7±0 .2 9) ,(38± 11)ng·L- 1降至 (1.1± 0 .3)分 ,(0 .2±1.6 ) % ,(0 .3± 0 .11) ,(11± 3)ng·L- 1。CD2 +,CD4 +,CD8+,CD5 8+分子荧光强度亦有不同程度降低 ,而mPEG BTC修饰对淋巴细胞的形态、寿命及遗传物质无明显改变。结论 mPEG BTC修饰可阻断HLA介导的特异性免疫反应 ,降低淋巴细胞的增殖、抗原分泌能力及分泌 ,但对其形态、结构、遗传物质及寿命无明显影响。
Objective To investigate the effect of mPEG BTC modified lymphocyte surface human leukocyte antigen (HLA) on cell injury. Methods The effects of mPEG BTC on the proliferation of lymphocytes were detected by MTT assay and unidirectional mixed lymphocyte culture. The morphological changes of lymphocytes were observed by electron microscopy. The levels of lymphocytes in lymphocytes were detected by lymphocyte transformation assay and IL- The proliferation, differentiation and secretion of cells were evaluated. CD molecules on lymphocytes were evaluated for antigen recognition of lymphocytes. Lymphocytes were preserved in vitro to measure their life span. Lymphocyte chromosomes were analyzed to evaluate the effect of mPEG on cytogenetic material. Results The results of lymphocyte cytotoxicity assay, lymphocyte transformation rate, lymphocyte relative transformation index and IL-2 secretion of modified lymphocytes were (8.0 ± 0), (3.6 ± 7.8)%, (1.0 7 ± 0.29), (38 ± 11) ng · L-1 decreased to (1.1 ± 0.3), (0.2 ± 1.6)%, (0.3 ± 0.11), (11 ± 3) ng · L -1. Fluorescence intensity of CD2 +, CD4 +, CD8 +, CD5 8+ molecules also decreased to some extent, while mPEG BTC modification had no obvious change on the morphology, life span and genetic material of lymphocytes. Conclusion mPEG BTC modification can block HLA-mediated specific immune response, reduce lymphocyte proliferation, antigen secretion and secretion, but have no significant effect on its morphology, structure, genetic material and life span.