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幽门螺杆菌感染蒙古沙鼠模型中对一种26千道尔顿蛋白质——烷基氢过氧化物还原酶的免疫反应。

Immune response to a 26-kDa protein, alkyl hydroperoxide reductase, in Helicobacter pylori-infected Mongolian gerbil model.

作者信息

Yan J, Kumagai T, Ohnishi M, Ueno I, Ota H

机构信息

Department of Laboratory Medicine, Shinshu University School of Medicine, Matsumoto, Nagano, Japan.

出版信息

Helicobacter. 2001 Dec;6(4):274-82. doi: 10.1046/j.1523-5378.2001.00038.x.

Abstract

BACKGROUND

The host immune response is thought to play an important role in the outcome of Helicobacter pylori infection. The successful development of the H. pylori-infected Mongolian gerbil model that mimics human disease has enabled study of the antibody response against H. pylori antigens.

MATERIALS AND METHODS

Serum samples from ulcer and carcinogenesis models of H. pylori-infected gerbils were used to screen for H. pylori antigens that cause a humoral immune response in the infected hosts. H. pylori alkyl hydroperoxide reductase (AhpC) is one such antigen on which we report here. The tsaA gene encoding AhpC was amplified by PCR from H. pylori ATCC 43504 strain, cloned into pMAL(TM)-c2 expression vector and expressed in Escherichia coli. Maltose-binding protein fusion protein (MBP-AhpC) was purified by a MBP affinity column. Using purified recombinant AhpC protein as an antigen, the antibody response and changes of antibody levels against AhpC in the gerbil models were studied by Western blotting and ELISA.

RESULTS

Antibody against AhpC was negative in the early stages of infection, and became positive in the gerbils with the emergence of gastric diseases such as chronic active gastritis, gastric ulcer and gastric cancer. The antibody levels (ELISA) increased gradually over time and were higher in gerbils with gastric ulcer than that in gerbils without ulcers.

CONCLUSIONS

Use of the gerbil model that mimics human H. pylori infection is likely to provide insights into the role of H. pylori-specific antigens possibly related to the subsequent development of gastric diseases.

摘要

背景

宿主免疫反应被认为在幽门螺杆菌感染的结局中起重要作用。成功建立模仿人类疾病的幽门螺杆菌感染蒙古沙鼠模型,使得针对幽门螺杆菌抗原的抗体反应研究成为可能。

材料与方法

使用幽门螺杆菌感染沙鼠的溃疡和致癌模型的血清样本,筛选在感染宿主中引发体液免疫反应的幽门螺杆菌抗原。幽门螺杆菌烷基过氧化氢还原酶(AhpC)就是我们在此报告的一种此类抗原。通过PCR从幽门螺杆菌ATCC 43504菌株中扩增编码AhpC的tsaA基因,克隆到pMAL(TM)-c2表达载体中,并在大肠杆菌中表达。麦芽糖结合蛋白融合蛋白(MBP-AhpC)通过MBP亲和柱纯化。以纯化的重组AhpC蛋白为抗原,通过蛋白质印迹法和酶联免疫吸附测定法研究沙鼠模型中针对AhpC的抗体反应及抗体水平变化。

结果

感染早期针对AhpC的抗体为阴性,在出现慢性活动性胃炎、胃溃疡和胃癌等胃部疾病的沙鼠中变为阳性。抗体水平(酶联免疫吸附测定法)随时间逐渐升高,有胃溃疡的沙鼠中的抗体水平高于无溃疡的沙鼠。

结论

使用模仿人类幽门螺杆菌感染的沙鼠模型,可能有助于深入了解幽门螺杆菌特异性抗原在胃部疾病后续发展中可能发挥的作用。

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