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In Vitro Assay of Bacterial Adhesion onto Mammalian Epithelial Cells
Published on: May 16, 2011
Assessment of Pasteurella multocida A Lipopolysaccharide, as an Adhesin in an In Vitro Model of Rabbit Respiratory
Carolina Gallego1, Stefany Romero2, Paula Esquinas3
1Laboratory of Veterinary Pathology, Universidad de Ciencias Aplicadas y Ambientales, Calle 222 No. 55-37, Bogotá, Colombia; Faculty of Science, Pontificia Universidad Javeriana, Carrera 7 No. 43-82, Bogotá, Colombia.
Abstract:
The role of the P. multocida lipopolysaccharide (LPS) as a putative adhesin during the early stages of infection with this bacterium in the respiratory epithelium of rabbits was investigated. By light microscopy and double enzyme labeling of nasal septa tissues, the amount of bacteria attached to the respiratory epithelium and the amount of LPS present in goblet cells at different experimental times were estimated. Transmission electron microscopy (TEM) and LPS labeling with colloidal gold particles were also used to determine the exact location of LPS in the cells. Septa that were challenged with LPS of P. multocida and 30 minutes later with P. multocida showed more adherent bacteria and more severe lesions than the other treatments. Free LPS was observed in the lumen of the nasal septum, forming bilamellar structures and adhering to the cilia, microvilli, cytoplasmic membrane, and cytoplasm of epithelial ciliated and goblet cells. The above findings suggest that P. multocida LPS plays an important role in the process of bacterial adhesion and that it has the ability of being internalized into host cells.
Insights
Pasteurella multocida lipopolysaccharide (LPS) acts as an adhesin, promoting bacterial attachment to respiratory tissues. This LPS can also be internalized by host cells, contributing to infection.
Area of Science:
- Microbiology
- Immunology
- Pathogenesis
Background:
- * *Pasteurella multocida* ( *P. multocida* ) is a significant respiratory pathogen.
- * Lipopolysaccharide (LPS) is a key component of Gram-negative bacterial outer membranes.
- * The role of *P. multocida* LPS in early infection stages requires further elucidation.
Purpose of the Study:
- * To investigate the role of *P. multocida* LPS as an adhesin in respiratory infections.
- * To determine the localization and interaction of LPS with host respiratory epithelial cells.
Main Methods:
- * Light microscopy and double enzyme labeling of rabbit nasal septa tissues.
- * Transmission electron microscopy (TEM) with colloidal gold-labeled LPS.
- * Quantitative assessment of bacterial adhesion and LPS presence.
Main Results:
- * *P. multocida* LPS pre-treatment increased bacterial adherence and lesion severity.
- * Free LPS was found in nasal lumen, forming bilamellar structures.
- * LPS adhered to and was internalized by epithelial cells (ciliated and goblet).
Conclusions:
- * *P. multocida* LPS significantly contributes to bacterial adhesion to host cells.
- * LPS internalization by host cells is a potential mechanism in infection.
- * LPS plays a crucial role in the early pathogenesis of *P. multocida* infections.
