JeanMarie Houghton (Editor)

  • Hardcover: 268 pages
  • Publisher: --; 2013 edition (September 27, 2012)
  • Language: English
  • ISBN-10: 1627030042
  • ISBN-13: 978-1627030045

  • Presents methods and protocols commonly used to study Helicobacter
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice form the experts

Identifying Helicobacter infection as the leading cause of peptic ulcer disease and gastric cancer has dramatically altered the treatment of these disease states. Over the last several decades, scientists have come to understand that the interplay between the bacteria, the host, and the environment all contribute to the clinical outcome of infection. In Helicobacter Species: Methods and Protocols, expert researchers in the field detail many of the methods and which are now commonly used to study Helicobacter infection. These include protocols and methods that have evolved over time, and standards across the field have been established which are essential for optimal outcomes and to allow comparison of data across different laboratories. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Helicobacter Species: Methods and Protocols seeks to aid scientists in further study of this crucially important research into Helicobacter research.

Preface . . . v
Contributors. . . ix
1 Introduction . . .1
2 Helicobacter pylori: An Overview . . .7
3 Perspectives on Methodology for In Vitro Culture of Helicobacter pylori . . .11
4 Successful Culture Techniques for HelicobacterSpecies: General Culture Techniques for Helicobacter pylori. . .17
5 Successful Culture Techniques for Helicobacter Species: Establishing H. pyloriCultures from Infected Rodents. . .29
6 Successful Culture Techniques for Helicobacter Species: Verification of HelicobacterIdentity Using 16S rRNA Gene Sequence Analysis... 37
7 The Helicobacter pylori cagPathogenicity Island . . .41
8 Genetic Manipulation of a Naturally Competent Bacterium,Helicobacter pylori. . .51
9 A Method for Short-Term Culture of Human Gastric Epithelial Cells to Study the Effects of Helicobacter pylori. . .61
10 Cell Culture-Based Assays to Test for Bacterial Adherence and Internalization ... 69
11 Cell Culture Assays to Evaluate Bacterial Toxicity and Virulence . . .77
12 Rodent Models of HelicobacterInfection, In fl ammation and Disease . . .89
13 Bacterial Culture and Inoculation of Mice (Simple Infection) . . .99
14 Adoptive Transfer of Splenocytes to Immunocompromised Mice. . .109
15 Isolation of Gastric Lamina Propria Leukocytes . . .113
16 Delayed-Type Hypersensitivity Determination. . .117
17 Necropsy, Blood, Tissue Collection and mRNA Isolation for Detection of Host Cytokine Gene Expression . . .119
18 Animal Models of Helicobacter-Induced Disease: Methods to Successfully Infect the Mouse . . .131
19 Verifying and Quantifying Helicobacter pyloriInfection Status of Research Mice. . .143
20 Mouse Models of Helicobacter-Induced Gastric Cancer: Use of Cocarcinogens . . .157
21 Gastric Helicobacterspp. in Animal Models: Pathogenesis and Modulation by Extragastric Coinfections. . . 175
22 Histologic Scoring of Gastritis and Gastric Cancer in Mouse Models . . .189
23 Innate Immune Responses to Helicobacter pyloriInfection: An Overview. . . . . 205
24 Methods for In Vivo and In Vitro Analysis of Innate Immune Responses to Helicobacter pyloriInfection. . . 209
25 Techniques for Following Labeled Cells In Vivo: Use of X/Y FISH, Techniques to Optimize Fluorescent Detection, and Beta-Galactosidase Detection . . . 227
26 In Vivo Measurement of Helicobacter pyloriInfection . . . 239
Index. . . 257

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