抗P-选择素功能域单抗体外抗黏附作用的实验研究

目的 观察针对P-选择素的抗P-选择素ktin-EGF功能域单抗(PsL-EGFmAh)体外抗黏附功能。方法 常规培养或制备人脐静脉内皮细胞(ECV-304)、中性粒细胞及肾小管上皮细胞(HK2细胞)，并对ECV-304和HK2细胞行炎症因子TNF-d刺激，然后观察和比较经PsL-EGFmAh和抗P-选择素全长单抗以及配体拮抗剂Ⅳ一去硫酸肝素处理前后，中性粒细胞与ECV-304细胞黏附变化，以及HK2细胞的黏附分子(P-选择素)和共刺激分子(CD80)表达情况。结果 经PsL-EGFmAb等三种药物处理后，中性粒细胞与受刺激ECV-304细胞黏附量随药物浓度均下降，受刺激后HK2细胞的P-选择素表达及CD 80mRNA转录水平也均受到抑制，其中尤以PsL-EGFmAh抗黏附抑制效应更明显。结论 抗P-选择素功能域单抗较其他两种药物更具体外抗黏附靶向抑制效应，为进一步研究针对P-选择素抗黏附作用提供了基础。

Anti-adhesive function of anti-P-selectin lectin-EGF domain monoclonal antibody in vitro

Objective To investigate the function of anti-P-selectin lectin-EGF domain monoclonal antibody ( PsL-EGFmAb) in vitro. Methods Human umbilical vein endothelial cells ( ECV-304 ), neutrophils and renal tubular epithelial cells ( HK2 cells) were cultured and prepared. ECV-304 and HK2 cells were stimulated by TNF-a. We processed these cells with PsL-EGFmAb, anti-P-selectin monoclonal antibody ( PsmAb ) and N-desulfated heparin. Then we observed the adhesion between neutrophils and ECV-304, the expression of P-selectin and CD80 in HK2 cells, and compared this effect. Results The adhesion between neutrophils and ECV-304 stimulated was decreased according to the concentration of agents in three treatment groups than TNF-a group. The expression of P-selectin and CD80 mRNA in HK2 cells was suppressed obviously. The anti-adhesive effect of PsL-EGFmAb was stronger than others. Conclusion PsL-EGFmAb had marked anti-adhesive function in vitro, and its effect was stronger than PsmAb and N-Desulfated heparin, which is the basis for further anti-adhesive study.