Host Resistance Model to an Intracellular Pathogen

Ling Cao1

1 Dartmouth‐Hitchcock Medical Center, Lebanon, New Hampshire
Publication Name:  Current Protocols in Toxicology
Unit Number:  Unit 18.10
DOI:  10.1002/0471140856.tx1810s27
Online Posting Date:  March, 2006
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Abstract

This unit describes a model for the determination of host resistance in mice using a Gram‐positive intracellular bacterium, Listeria monocytogenes (LM), and discusses its broad use in immunotoxicological studies. The provides detailed procedures for LM infection and sample analysis, including preparation and infection of mice with LM, recording of sickness behaviors in infected mice, and determination of viable LM numbers in the tissues from infected animals. The protocol also describes use of serum and tissue homogenates for assessment of cytokines. Additionally, background information on LM and LM infection is provided for better understanding and utilization of this model. The LM infection model is useful for the initial screening of possible modulation of immune responses, particularly innate immunity and type‐1 cell‐mediated immunity (CMI), by environmental factors, as well as for further investigation of the underlying mechanisms of environmental factor–induced changes in immunity.

Keywords: Listeria monocytogenes (LM); Intracellular pathogen; Host resistance

     
 
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Table of Contents

  • Strategic Planning
  • Basic Protocol 1: Determination of Host Resistance to LM Infection in Mice
  • Reagents and Solutions
  • Commentary
  • Literature Cited
     
 
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Materials

Basic Protocol 1: Determination of Host Resistance to LM Infection in Mice

  Materials
  • 100‐mm 5% (v/v) horse or sheep blood agar plates (BD Biosciences; store at 4°C)
  • Frozen (−80°C) stock of virulent LM, with known concentration (Campbell et al., ; Harding, )
  • Saline (0.9% w/v NaCl)
  • Mice to be infected (see )
  • 70% ethanol
  • Tissue lysis buffer (see recipe)
  • Nonidet P‐40 (NP‐40)
  • Permanent marker, dark color
  • 12 × 75–mm culture tubes (polypropylene or polystyrene culture tubes, or autoclaved borosilicate glass tubes), sterile
  • Noncoated or silicone‐coated Vacutainer blood collection tube (BD Biosciences)
  • Balance accurate to 0.01 g to weigh the mice
  • 1‐ml syringe with 27‐G needle (one per mouse preferred; for intravenous injection of the live mice)
  • Sealable plastic bags to accommodate petri dishes
  • Tissue homogenizer (Potter‐Elvejem or Polytron‐type) and appropriate homogenizer tubes
  • 1‐ml syringe with 25‐G needle (one per mouse preferred; for drawing blood from mice by cardiac puncture)
  • Refrigerated centrifuge
  • Additional reagents and equipment for intravenous (i.v.) injection of the mouse via the tail vein (Donovan and Brown, ) and euthanasia of the mouse by CO 2 asphyxiation (Donovan and Brown, ), blood collection from the mouse via cardiac puncture (Donovan and Brown, ), and ELISA (optional; unit 18.7).
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Figures

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Literature Cited

Literature Cited
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