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目的 鉴定抗 L PS m Ab对不同种属细菌及其 L PS的交叉反应性和保护活性 .方法 以间接 EL ISA,Western印迹试验鉴定 m Ab对不同种属细菌及其 L PS的交叉反应性 ;检测不同浓度 m Ab对 L PS诱导的淋巴细胞分泌 TNF- α和IL- 6的抑制作用 ;以大肠埃希菌、铜绿色假单胞菌、鼠伤寒沙门菌建立小鼠菌血症模型 ,鉴定 m Ab的保护作用 .结果 在EL ISA试验中 ,抗 L PS m Ab C3A2和 H3D3可与鉴定所用的18株革兰阴性菌产生交叉反应 ;在 Western印迹试验中可与鉴定所用的全部 7个种属的 L PS产生交叉反应 ;随 m Ab浓度的提高 ,L PS诱导淋巴细胞分泌 TNF- α和 IL- 6的作用受到抑制 ;在动物保护试验中 ,相对于生理盐水对照组 ,存活率提高30 %～ 6 0 % .结论 抗 L PS m Ab C3A2和 H3D3不仅对众多革兰阴性菌 L PS具有广泛交叉反应性 ,而且具有高保护活性
Objective To identify the cross-reactivity and protective activity of anti-L PS m Ab against different species of bacteria and their L PS.Methods The cross-reactivity of m Ab to bacteria and their L PS was evaluated by indirect ELISA and Western blotting. The inhibitory effect of different concentrations of m Ab on the secretion of TNF-α and IL-6 by LPS-induced lymphocytes was tested. The model of bacteremia was established by using Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium. m Ab Results Results In the EL ISA assay, anti-LPS m Ab C3A2 and H3D3 cross-reacted with 18 Gram-negative bacteria identified for use in identification; all 7 species used in the Western blotting assay L PS produced a cross-reaction; with the increase of m Ab concentration, LPS-induced lymphocyte secretion of TNF-α and IL-6 was inhibited in animal protection experiments, compared with the saline control group, the survival rate increased by 30 % ~ 60% .CONCLUSION Anti-L PS m Ab C3A2 and H3D3 not only have extensive cross-reactivity with many gram-negative bacteria L PS, but also have high protective activity