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A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening
Published on: January 17, 2014
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Experimental selection of long-term intracellular mycobacteria
Cristina L Vázquez1, Thomas R Lerner, Bahram Kasmapour
1Research Group Phagosome Biology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124, Braunschweig, Germany.
Cellular Microbiology
|May 1, 2014
Summary
Mycobacterium bovis BCG adapts to long-term host cell residence by altering its metabolism and lipid synthesis. These adaptations enhance bacterial survival during re-infection, offering insights into chronic intracellular infections.
Area of Science:
- Microbiology
- Infectious Diseases
- Bacterial Pathogenesis
Background:
- Intracellular bacteria can establish chronic infections lasting decades.
- Adaptations of bacteria during long-term host cell residence are poorly understood.
Purpose of the Study:
- To investigate the adaptations of mycobacteria during a long-term intracellular lifestyle.
- To identify bacterial traits crucial for establishing persistent cellular infections.
Main Methods:
- Continuous culture of Mycobacterium bovis BCG within macrophages.
- Analysis of changes in phenolic glycolipids (PGL) synthesis, carbon source utilization, and lipid accumulation.
- Assessment of bacterial survival during re-infection in macrophages and mice.
- Gene expression analysis of stress- and survival-related genes.
Main Results:
- Adapted Mycobacterium bovis BCG exhibited impaired PGL synthesis.
- Enhanced utilization of glucose as a carbon source was observed.
- Accumulation of neutral lipids occurred in adapted bacteria.
- Increased survival rates in macrophages and mice during re-infection correlated with these changes.
- Specific stress- and survival-related genes were upregulated.
Conclusions:
- Long-term intracellular residence induces significant adaptive changes in mycobacteria.
- Altered PGL synthesis, improved glucose metabolism, and neutral lipid accumulation are key traits for persistent infection.
- These findings provide a framework for understanding bacterial adaptation in fluctuating intracellular environments.
