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A Precise Pathogen Delivery and Recovery System for Murine Models of Secondary Bacterial Pneumonia
Published on: September 21, 2019
A Precise Pathogen Delivery and Recovery System for Murine Models of Secondary Bacterial Pneumonia
Timothy R Borgogna1, Adrian Sanchez-Gonzalez2, Kelly Gorham2
1Department of Microbiology and Immunology, Montana State University.
Abstract:
Secondary bacterial pneumonias following influenza infections consistently rank within the top ten leading causes of death in the United States. To date, murine models of co-infection have been the primary tool developed to explore the pathologies of both the primary and secondary infections. Despite the prevalence of this model, considerable discrepancies regarding instillation procedures, dose volumes, and efficacies are prevalent among studies. Furthermore, these efforts have been largely incomplete in addressing how the pathogen may be directly influencing disease progression post-infection. Herein we provide a precise method of pathogen delivery, recovery, and analysis to be used in murine models of secondary bacterial pneumonia. We demonstrate that intratracheal instillation enables an efficient and accurate delivery of controlled volumes directly and evenly into the lower respiratory tract. Lungs can be excised to recover and quantify the pathogen burden. Following excision of the infected lungs, we describe a method to extract high quality pathogen RNA for subsequent transcriptional analysis. This procedure benefits from being a non-surgical method of delivery without the use of specialized laboratory equipment and provides a reproducible strategy to investigate pathogen contributions to secondary bacterial pneumonia.
Insights
This study presents a standardized method for studying secondary bacterial pneumonia in mice. It details precise pathogen delivery and analysis for better understanding bacterial roles in disease progression.
Area of Science:
- Microbiology
- Immunology
- Pathology
Background:
- Secondary bacterial pneumonias post-influenza are a major cause of mortality.
- Murine co-infection models are crucial but lack standardized procedures.
- Existing models inadequately address pathogen-specific disease contributions.
Purpose of the Study:
- To establish a precise and reproducible method for pathogen delivery, recovery, and analysis in murine models of secondary bacterial pneumonia.
- To overcome discrepancies in current co-infection models.
- To facilitate investigation into pathogen-driven disease progression.
Main Methods:
- Intratracheal instillation for accurate, even delivery of pathogens to the lower respiratory tract.
- Lung excision for pathogen burden quantification.
- High-quality pathogen RNA extraction from excised lungs for transcriptional analysis.
- Non-surgical procedure requiring no specialized equipment.
Main Results:
- Demonstrated efficient and accurate pathogen delivery via intratracheal instillation.
- Established a reproducible method for pathogen recovery and RNA extraction.
- Validated a non-surgical approach for reproducible murine pneumonia studies.
Conclusions:
- The described method offers a reproducible strategy for investigating pathogen contributions to secondary bacterial pneumonia.
- This standardized approach enhances the utility of murine models for co-infection research.
- Facilitates deeper understanding of bacterial roles in post-influenza pneumonia pathogenesis.
