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Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions
Shrilakshmi Hegde1, Cordula Gabriel2, Martin Kragl1
1Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine Vienna, Veterinaerplatz 1, A-1210 Vienna, Austria;
Abstract:
Appropriate infection models are imperative for the understanding of pathogens like mycoplasmas that are known for their strict host and tissue specificity, and lack of suitable cell and small animal models has hindered pathogenicity studies. This is particularly true for the economically important group of ruminant mycoplasmas whose virulence factors need to be elucidated for designing effective intervention strategies. Mycoplasma agalactiae serves as a useful role model especially because it is phylogenetically very close to M. bovis and causes similar symptoms by as yet unknown mechanisms. Here, we successfully prepared and characterized four different primary sheep cell lines, namely the epithelial and stromal cells from the mammary gland and uterus, respectively. Using immunohistochemistry, we identified vimentin and cytokeratin as specific markers to confirm the typical cell phenotypes of these primary cells. Furthermore, M. agalactiae's consistent adhesion and invasion into these primary cells proves the reliability of these cell models. Mimicking natural infections, mammary epithelial and stromal cells showed higher invasion and adhesion rates compared to the uterine cells as also seen via double immunofluorescence staining. Altogether, we have generated promising in vitro cell models to study host-pathogen interactions of M. agalactiae and related ruminant pathogens in a more authentic manner.
Insights
Researchers developed new sheep cell models to study Mycoplasma agalactiae, a pathogen affecting ruminants. These models mimic natural infections, aiding the understanding of mycoplasma pathogenicity and virulence factors for better intervention strategies.
Area of Science:
- Veterinary Microbiology
- Cell Biology
- Infectious Diseases
Background:
- Mycoplasma infections in ruminants pose significant economic challenges.
- Understanding mycoplasma virulence factors requires suitable host models.
- Mycoplasma agalactiae is a relevant model organism due to its similarity to Mycoplasma bovis.
Purpose of the Study:
- To develop and characterize primary sheep cell lines for studying ruminant mycoplasma infections.
- To establish reliable in vitro models for investigating Mycoplasma agalactiae host-pathogen interactions.
- To provide a platform for elucidating virulence factors of economically important ruminant mycoplasmas.
Main Methods:
- Preparation and characterization of four primary sheep cell lines (mammary and uterine epithelial and stromal cells).
- Immunohistochemistry using vimentin and cytokeratin as specific cell markers.
- Assessment of Mycoplasma agalactiae adhesion and invasion in the developed cell models via immunofluorescence staining.
Main Results:
- Successfully established and validated four distinct primary sheep cell lines.
- Confirmed cell phenotypes using specific markers (vimentin and cytokeratin).
- Demonstrated consistent adhesion and invasion of Mycoplasma agalactiae in mammary and uterine cells, with higher rates in mammary cells.
Conclusions:
- Developed reliable in vitro cell models for studying Mycoplasma agalactiae.
- These models accurately mimic natural infection dynamics, showing differential pathogen interaction with mammary and uterine cells.
- The established cell lines offer a promising tool for future research on ruminant mycoplasma pathogenicity and intervention strategies.
