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目的 建立可特异性识别IL - 8抗原并具有拮抗IL - 8生物学活性的单抗 ,观察抗体对炎症性损伤的保护作用。方法 采用常规方法免疫和细胞融合 ,经间接ELISA检测和有限稀释亚克隆、筛选 ,建立稳定分泌抗人IL - 8的杂交瘤细胞株。通过趋化阻断实验 ,观察抗体在体外拮抗IL - 8生物学活性的作用 ;以家兔为模型 ,通过皮肤注射LPS诱导局部产生炎症反应 ,同时静脉注射IL - 8抗体 ,观察抗体对炎症性损伤的保护作用。结果 经筛选获得 3株稳定分泌IL - 8单抗的杂交瘤细胞株 ,一株为IgG1(SI98) ,另两株为IgM (SI96、SI97)。这 3株单抗在体外具有阻断IL - 8对中性粒细胞的趋化作用 ;在体内 ,皮肤局部注射LPS产生的炎症反应 ,导致大量中性粒细胞的浸润 ,体内预先注射IL - 8抗体后 ,可明显阻断中性粒细胞在炎症部位的浸润。结论 3株单抗均可作为拮抗IL - 8生物学功能的拮抗剂 ,阻断炎症反应过程中中性粒细胞的浸润。
Objective To establish a monoclonal antibody that can specifically recognize IL - 8 antigen and antagonize the biological activity of IL - 8, and to observe the protective effect of antibody against inflammatory injury. Methods Immunization and cell fusion were performed by conventional methods. Subclones were screened by indirect ELISA and limited dilution to establish a hybridoma cell line secreting anti - human IL - 8. The effect of blocking the biological activity of IL - 8 in vitro was observed by chemoattractant blocking assay. The rabbit model was induced by local injection of LPS, and the IL - 8 antibody was also injected intravenously. Injury protection. Results Three hybridoma cell lines stably secreting IL - 8 mAb were screened. One was IgG1 (SI98) and the other two were IgM (SI96, SI97). The three monoclonal antibodies blocked the chemotactic effect of IL - 8 on neutrophils in vitro. Inflammatory reaction induced by local injection of LPS into the skin in vivo resulted in a large number of neutrophil infiltration. In vivo, pretreatment with IL - 8 Antibodies, can significantly block neutrophil infiltration in the inflammatory sites. Conclusion All three monoclonal antibodies can antagonize the biological function of IL - 8 and block the infiltration of neutrophils during inflammation.