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JoVE Journal
Immunology and Infection
Use of the EpiAirway Model for Characterizing Long-term Host-pathogen Interactions
Use of the EpiAirway Model for Characterizing Long-term Host-pathogen Interactions
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Use of the EpiAirway Model for Characterizing Long-term Host-pathogen Interactions

Use of the EpiAirway Model for Characterizing Long-term Host-pathogen Interactions

Full Text
12,156 Views
08:12 min
September 2, 2011

DOI: 10.3791/3261-v

Dabin Ren1, Dayle A. Daines1

1Division of Basic Medical Sciences,Mercer University School of Medicine

Overview

This method allows characterization of extended bacterial co-culture with EpiAirways, a biologically relevant in vitro model using primary human respiratory epithelial tissue. The approach can be utilized with any microbe suitable for long-term co-culture.

Key Study Components

Area of Science

  • Microbiology
  • Respiratory Biology
  • In Vitro Models

Background

  • Nontypeable Hemophilus influenza (NTHI) is a significant pathogen in respiratory infections.
  • Understanding bacterial interactions with respiratory epithelial tissues is crucial for developing treatments.
  • The EpiAirways model mimics the human airway environment.
  • Long-term co-culture studies can provide insights into bacterial survival and behavior.

Purpose of Study

  • To perform long-term co-culture of NTHI with primary human respiratory epithelial tissues.
  • To evaluate the survival of NTHI within the tissues over time.
  • To assess strain-specific characteristics of NTHI survival.

Methods Used

  • Inoculation of the apical surface of epithelial tissues with NTHI.
  • Washing inserts every 24 hours with pre-warmed DPBS to mimic mucus flow.
  • Harvesting tissues at desired time points for analysis.
  • Determining survival of total cell-associated or internalized NTHI.

Main Results

  • Insights into long-term infections with NTHI were gained.
  • Viable bacteria counts were used to evaluate NTHI survival characteristics.
  • The method demonstrated the ability to assess bacterial behavior in a relevant model.
  • Findings could inform future research on respiratory infections.

Conclusions

  • This method provides a valuable tool for studying bacterial co-culture with human respiratory tissues.
  • It enhances understanding of NTHI interactions with epithelial cells.
  • The approach can be adapted for other microbes in similar studies.

Frequently Asked Questions

What is the EpiAirways model?
The EpiAirways model is an in vitro system that mimics human respiratory epithelial tissue grown at the air-liquid interface.
How is NTHI introduced to the epithelial tissues?
NTHI is inoculated onto the apical surface of the tissues to allow adherence and entry into the cells.
What is the significance of washing the inserts?
Washing the inserts every 24 hours helps to mimic normal mucus flow and removes waste products and debris.
What outcomes are measured in this study?
The study measures the survival of total cell-associated or internalized NTHI within the tissues over time.
Can this method be used for other microbes?
Yes, the approach can be adapted for any microbe suitable for long-term co-culture.
What insights can be gained from this method?
The method provides insights into long-term infections and bacterial behavior in a relevant biological model.

This method allows characterization of extended bacterial co-culture with EpiAirways, primary human respiratory epithelial tissue grown at the air-liquid interface, a biologically relevant in vitro model. The approach can be used with any microbe that is amenable to long-term co-culture.

The overall goal of the following experiment is to perform long-term co-culture of Nontypeable Hemophilus influenza or NTHI with primary human respiratory epithelial tissues using the epi airway model. This is achieved by inoculating the apical surface with NTHI to allow the bacteria to adhere to and enter the tissues as a second step. The inserts are washed every 24 hours with prewarm DPBS to mimic normal mucus flow and to remove mucin waste products and any cellular debris.

Next, the tissues are harvested at the desired time point and the survival of either total cell associated or internalized NTHI is determined. Finally, with this method, strain specific characteristics of NTHI survival within the tissues over time based on viable bacteria counts can be evaluated. Though this method can provide insight into long-term infections with nontypeable hemophilus influenza.

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