IL-8单抗对炎症性中性粒细胞浸润的阻断作用

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。

Inhibition Effect of Monoclonal Antibody against Interleukin-8 on LPS Induced Neutrophil Infiltration

Objective To establish a monoclonal antibody against Interleukin-8 (IL-8) and investigate its roles in inflammatory injury.Methods Balb/c mice were immunized with human IL-8 by routine methods. IL-8 hybridoma was screened by an indirect Enzyme linked Immuno-Sorbed Assay (ELISA). Interleukin-8 has chemotactic activity on PMN in vitro, and the anti-chemotactic activity of the monoclonal antibody against IL-8 was examined using a modified-multiwell Boyden chamber system. To investigate its roles when inflammation, LPS was injected into rabbit skin for inducing inflammation, neutrophil infiltration was examinated by routine histological method after inhibited by IL-8 antibody.Results Three anti-IL-8 mAbs were obtained, SI96 and SI97 are IgM mAbs, SI98 belongs to IgG 1. Three mAbs have the blocking effect on IL-8 in vitro, and the chemotactic effect of IL-8 was completely inhibited when the concentration of antibody was increased. After injecting LPS 4 h, lots of neutrophils were infiltrated in inflammatory skin, but obviously decreased when protected by IL-8 antibody.Conclusion Three IL-8 mAbs have the blocking effect against IL-8 in vivo, which can be used to inhibit inflammatory injury in vivo.