实验基础

In vitro studies of Helicobacter pylori pathogenesis mostly rely on the use of tumor-derived cell lines. Although invaluable, tumor cell lines are not representative of the normal cell physiology. Thus, the use of primary gastric epithelial cell cultures provides an important tool for in ...

Genetic manipulation of Helicobacter pylori facilitates characterization and functional analysis of individual H. pylori genes. This chapter discusses the methods involved in H. pylori chromosomal DNA isolation, mutagenesis of individual genes, and natural transformat ...

The cag pathogenicity island is a well-characterized virulence determinant. It is composed of 32 genes that encode a type IV bacterial secretion system and is linked with a more severe clinical outcome. The following chapters will explore the manipulation of bacterial factors in order to un ...

This chapter describes protocols for the verification of putative Helicobacter species’ identities using 16S rRNA gene sequence analysis.

Half of the world’s population is persistently infected with Helicobacter pylori. The chronicity of this infection ultimately elicits clinical manifestations ranging from gastritis and peptic ulcers to adenocarcinoma and MALT lymphoma. Laboratory research following the i ...

Over the past 25 years, a variety of methods have been developed for culture of Helicobacter pylori in vitro. H. pylori is a capnophilic and microaerophilic organism that is typically cultured using complex culture media. Analysis of H. pylori growth in chemically defined media has provided in ...

Infection with Helicobacter pylori is directly responsible for substantial morbidity and mortality worldwide. This ubiquitous organism causes disease through the interaction of multiple factors including bacterial factors, host immune responses, and environmental f ...

Helicobacter pylori is a well-recognized gastroduodenal pathogen (National Institute of Health Consensus Conference, JAMA 272:65–9, 1994) and a class I carcinogen (International Agency for Research on Cancer, IARC Monograph on the Evaluation of Carcinogenic Risk to Humans 61:17 ...

The redistribution and trafficking patterns of cells to different anatomic sites throughout the body is important during cancer development and metastasis. Interest in the origin and fate of gastric cancer stem cells has recently arisen, as it may explain the underlying mechanism of can ...

It is estimated that half of the world’s population is infected by Helicobacter pylori (H. pylori) (Polk and Peek, Nat Rev Cancer 10:403–414, 2010; Peek et al., Physiol Rev 90:831–858, 2010). Following infection, H. pylori induces a chronic innate immune response that is thought to contribute to gastr ...

Innate immune receptors detect Helicobacter pylori infection and trigger downstream signaling events that result in the production of cytokines and interferon-β. This chapter gives an overview of the receptors and their roles in responding to H. pylori infection and details the down ...

Histopathology is a defining endpoint in mouse models of experimental gastritis and gastric adenocarcinoma. Presented here is an overview of the histology of gastritis and gastric cancer in mice experimentally infected with Helicobacter pylori or H. felis. A modular histopatholo ...

Animal models are used to study complex host, microbial, and environmental influences associated with gastric Helicobacter infection. Evidence that gastric helicobacters are pathogenic in animals first came from ferrets. Felids, nonhuman primates, and many other species also h ...

The human pathogen Helicobacter pylori causes inflammation in the stomach of infected hosts, leading in some cases to the development of gastric cancer. Several mouse models have been developed to study Helicobacter-induced carcinogenesis with similarities to gastric adenoca ...

Mice used to model helicobacter gastritis should be screened by PCR prior to experimental dosing to confirm the absence of enterohepatic Helicobacter species (EHS) that colonize the cecum and colon of mice. Natural infections with EHS are common and impact of concurrent EHS infection on Hel ...

Animal models of microbial diseases in humans are an essential component for determining fulfillment of Koch’s postulates and determining how the organism causes disease, host response(s), disease prevention, and treatment. In the case of Helicobacter pylori, establishing an anim ...

Processing of tissue and blood must be done in a systematic and controlled fashion in order to optimize results and allow comparison of samples between experiments and between laboratories. Here we present our protocols for blood and tissue processing.

Delayed-type hypersensitivity responses in the skin (in the case of mice, in the foot pad) is used to assess cell-mediated immunity (CMI) in vivo. In the case of CMI to Helicobacter infection, the mice are given an injection of cultured Helicobacter organisms into the hind footpad, and induration is m ...

Immune cells recruited to the infected gastric mucosa can be isolated and used for a variety of purposes. Here we describe methods for the isolation and characterization of gastric lamina propria leukocytes.

Analysis of the immune response of mice to Helicobacter infection has been greatly aided by the use of various deficient mouse strains. Here we present protocols for reconstitution of immune-deficient mice with wild-type immune cells and protocols for analysis of the outcome.