抗氨基端脂多糖结合蛋白单链抗体的生物学活性研究  

Bioactivity of scFv antibody against human NH-lipopolysaccharide binding protein in vitro and in vivo

作　　者：章容[1] 王长松[1] 刘友生[1] 李红[1] 

机构地区：[1]第三军医大学西南医院病理科,重庆400038

出　　处：《第三军医大学学报》2005年第24期2406-2409,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目(30271342)~~

摘　　要：目的观察制备的抗NHLBP单链抗体的体内外生物学活性`。方法采用流式细胞术观察抗NHLBP单链抗体阻断LPS与U937细胞的结合能力;采用ELISA法检测抗体干预后不同时相点的烧伤合并内毒素血症小鼠血浆中炎性细胞因子TNFα、IL6含量的变化;同时观察小鼠部分内脏器官的病理形态学改变。结果经抗NHLBP单链抗体干预后,U937细胞表面的荧光强度明显下降。烧伤合并内毒素血症小鼠经scFv抗体干预后,与模型组相比肝、肺组织的炎性改变明显减轻、浸润的炎细胞减少;肾组织的炎性改变不明显;小鼠血浆中TNFα浓度在烧伤合并内毒素处理后即显著增高,模型组与对照组相比,差异有显著性(P<0.01,P<0.05),干预组与相应时相点的模型组相比,在最初3h内差异有显著性(P<0.05)。IL6含量在建模3h以后与对照组相比差异有显著性(P<0.01,P<0.05);干预后IL6的含量下降比较明显。结论抗人源化氨基端脂多糖结合蛋白的单链抗体对内毒素血症有一定的保护作用;进一步明确了LBP在内毒素生物学效应中的作用。Objective To observe the bioactivity of recombinant human scFv antibody against NH-lipopolysaccharide binding protein in vitro and in vivo. Methods KM mice, weighing 20 -25 g, were burnt on the back to the third degree covering 20% total body surface area. The mice in only burn group were intraperitoneally injected with 1 μg/g LPS and those in scFv-treated group with 1 μg/g LPS and 0.5 μg/g scFv after burn. The mice without any treatment served as controls. All mice were sacrificed in 1,3,6, 12, 24 h after all treatment and the whole blood, the tissues of liver, lung and kidney were collected. The serum concentration of TNF-α, IL-6 in burnt mice with endotoxemia was detected by ELISA and the pathological changes of liver, lung and kidney tissues were observed by light microscope. The inhibition of scFv antibody of different concentrations against NH-lipopolysaccharide binding protein on FITC-LPS binding with U937 cells was assayed by FCM. Results The histopathological changes of liver, lung and kidney in mice of scFv-treated group revealed that less inflammatory cell infiltration, less degenerated cells, weaker congestion in tissue as compared with control and only burnt mice. The serum concentration of TNF-α in only burnt mice increased significantly than that of the controls （P 〈 0.01 and P 〈 0.05 ） , and the serum level of TNF-α in scFv-treated group in 1 h and 3 h was significantly lower than that of only burnt group （P 〈 0.05 ）. The content of IL-6 in the only burnt mice was significantly higher than that of controls after 3 h （ P 〈 0.01 and P 〈 0.05 ）. The IL-6 level of scFv-treated group decreased significantly than that of only burnt group （ P 〈 0.01 ）. The fluorescence intensity on U937 cells was weaken after treated by the scFv antibody. Conclusion The recombinant scFv antibody against NHLBP plays a potential protective role in the endotoxemia. In addition, result also revealed that LBP plays a key role in the pathogesis of LPS.

关 键 词：NH—LBP 单链抗体 内毒素血症 生物学活性 

分 类 号：R341.6[医药卫生—基础医学] R392.11