特异性抗幽门螺杆菌卵黄抗体的制备及其抗菌作用。
Preparation of specific anti-Helicobacter pylori yolk antibodies and their antibacterial effects.
作者信息
Wang Baoning, Yang Jing, Cao Suizhong, Wang Hongren, Pan Xing, Zhu Jie, Zhou Yongjun, Gao Lizhen, Li Wanyi, Li Mingyuan
机构信息
Department of Microbiology, West China of Preclinical and Forensic Medicine, Sichuan University Chengdu 610041, China.
Department of Microbiology, West China of Preclinical and Forensic Medicine, Sichuan University Chengdu 610041, China ; Sichuan Vaccine Technology Co., Ltd. Chengdu 610041, Sichuan, China ; Department of Medical Microbiology, School of Preclinical Medicine, Hubei University of Medicine Shiyan 442000, China.
出版信息
Int J Clin Exp Pathol. 2014 Sep 15;7(10):6430-7. eCollection 2014.
OBJECTIVE
To study immunization procedures and preparation methods of specific IgY antibodies (IgY-Hp, IgY-IB) produced by hens immunized with Helicobacter pylori (Hp) bacterial antigen and recombinant Hp specific antigen IB, detect the inhibition effects on Hp growth and Hp urease activity, and study the effects of oral administration for treating Hp infection.
METHODS
By using recombinant cholera toxin subunit B (rCTB) as an adjuvant, hens received intramuscular injection immunization for continuous 7 times at an interval of 14 days. Then, the eggs were collected; IgY was purified.
RESULTS
On day 49 after hens were immunized, levels of two antibodies all reached 1:12800; after they were purified by Ammonium sulfate precipitation, their purity was over 80%. IgY-Hp could inhibit Hp growth and inhibit Hp urease activity; although in vitro, IgY-IB could not inhibit Hp growth but could inhibit Hp urease activity. The experiments in vivo found that when IgY-Hp or IgY-IB with sucralfate dual oral therapy was used to treat Hp infected mouse model, the cure rate all could reach 83.3%.
CONCLUSION
According to immunization procedure, high titer specific IgY antibody (1:12800) can be obtained in 49 days and its titer remains stable. Oral administration of the specific IgY antibodies in Hp infected mice can reach a cure rate of 83.3%, and the antibodies are expected to become new drugs and therapeutic methods of targeted therapy against Hp infection.
目的
研究用幽门螺杆菌（Hp）细菌抗原和重组Hp特异性抗原IB免疫母鸡所产生的特异性IgY抗体（IgY-Hp、IgY-IB）的免疫程序及制备方法，检测其对Hp生长及Hp尿素酶活性的抑制作用，并研究口服给药治疗Hp感染的效果。
方法
以重组霍乱毒素B亚单位（rCTB）为佐剂，母鸡每隔14天进行1次肌肉注射免疫，连续免疫7次。然后收集鸡蛋；纯化IgY。
结果
母鸡免疫后第49天，两种抗体水平均达到1:12800；经硫酸铵沉淀法纯化后，纯度均超过80%。IgY-Hp可抑制Hp生长并抑制Hp尿素酶活性；虽然在体外，IgY-IB不能抑制Hp生长，但可抑制Hp尿素酶活性。体内实验发现，当用IgY-Hp或IgY-IB联合硫糖铝口服治疗Hp感染小鼠模型时，治愈率均可达83.3%。
结论
按照免疫程序，49天可获得高滴度特异性IgY抗体（1:12800）且滴度保持稳定。对Hp感染小鼠口服特异性IgY抗体治愈率可达83.3%，有望成为抗Hp感染靶向治疗的新药及治疗方法。
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