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Initial phase of infection with Tyzzer's organism in cultured mouse hepatocytes
S Kawamura1, T Mitsuoka, K Fujiwara
1Department of Biomedical Science, Faculty of Agriculture, University of Tokyo, Japan.
Abstract:
The initial phase of infection with Tyzzer's organisms in cultured mouse hepatocytes was observed using indirect immunofluorescence (IF) and a plaque assay. The organisms adhered poorly to aldehyde- or acetone-fixed cells, but once adhered to host cells, whether methanol-fixed or unfixed, they were not removed by methanol or acetone. By the IF as well as the plaque assay, both of which discriminated intra- and extracellularly located organisms, the number of cell-associated organisms increased linearly up to 3 h post-inoculation. The number of intra-cellular organisms increased rapidly in the first 1 h, followed by linear increase at a much lower rate. Similar results were obtained by the plaque assay.
Insights
Tyzzer
Area of Science:
- Microbiology
- Hepatology
- Infectious Diseases
Background:
- Tyzzer's disease is a significant cause of mortality in various animal species.
- Understanding the initial stages of Tyzzer's organism infection is crucial for developing effective treatments.
- Cultured mouse hepatocytes provide a model system to study host-pathogen interactions.
Purpose of the Study:
- To investigate the initial phase of Tyzzer's organism infection in cultured mouse hepatocytes.
- To characterize the adherence and intracellular entry of Tyzzer's organisms.
- To quantify organism-host cell interactions over time.
Main Methods:
- Indirect immunofluorescence (IF) for visualizing organisms.
- Plaque assay for quantifying viable organisms.
- Use of fixed and unfixed cultured mouse hepatocytes.
Main Results:
- Tyzzer's organisms exhibited poor adherence to aldehyde- or acetone-fixed cells.
- Adhered organisms were resistant to methanol and acetone fixation.
- Cell-associated organism numbers increased linearly up to 3 hours post-inoculation.
- Intracellular organism numbers rapidly increased in the first hour, then showed a slower linear increase.
Conclusions:
- The study elucidates the early adherence and intracellular dynamics of Tyzzer's organisms in hepatocytes.
- Both IF and plaque assays are effective in differentiating intra- and extracellular organisms.
- Findings provide insights into the initial host-pathogen interactions during Tyzzer's disease.