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以超声震荡及高速离心法裂解空泡毒素基因阳性 (VacA+ )HP临床分离株 ;采用Hela细胞体外培养法分析此HP菌体蛋白的细胞毒活性及其免疫产卵母鸡制备的IgY抗体的保护作用。HP菌体抗原制备物经SDS PAGE显示至少有十余个蛋白区带 ,主要区带位于约 940 0 0和 6 70 0 0。以此HP菌体抗原包被的ELISA显示 ,球部溃疡患者血清IgG抗体滴度明显增高。体外培养中HP菌体蛋白制备物对Hela细胞具有明显的细胞毒活性 ,LD50 小于 12 μg/ml。以此抗原免疫产卵母鸡 ,提取的IgY抗体浓度依赖地阻断HP菌体蛋白的Hela细胞毒活性 ;提示局部应用抗体制剂做被动免疫 ,有可能成为治疗HP感染性疾病的有效方法
The vacuolar toxin gene-positive (VacA +) HP clinical isolates were lysed by ultrasonication and high-speed centrifugation. The cytotoxic activity of the HP bacterial protein and the protection of IgY antibodies prepared from the immunopositive hens were analyzed by Hela cell culture in vitro effect. HP mycoplasma antigen preparations showed at least a dozen protein bands by SDS PAGE with major bands at about 940 and 060. ELISA with HP cell antigen coated ELISA showed that serum IgG antibody titers in patients with ulcer were significantly increased. HP bacterial protein preparations in vitro has obvious cytotoxic activity on Hela cells with LD50 less than 12 μg / ml. The antigen was used to immunize the laying hens, and the extracted IgY antibody blocked the Hela cytotoxic activity of the HP bacterial protein in a concentration-dependent manner. It suggests that the local application of the antibody preparation as a passive immunization may be an effective method for treating HP infectious diseases