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Streptococcus suis interactions with the murine macrophage cell line J774: adhesion and cytotoxicity
Mariela Segura1, Marcelo Gottschalk
1Groupe de Recherche sur les Maladies Infectieuses du Porc (GREMIP) and Canadian Research Network on Bacterial Pathogens of Swine, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada.
Abstract:
Streptococcus suis capsular type 2 is an important etiological agent of swine meningitis, and it is also a zoonotic agent. Since one hypothesis of the pathogenesis of S. suis infection is that bacteria enter the bloodstream and invade the meninges and other tissues in close association with mononuclear phagocytes, the objective of the present study was to evaluate the capacity of S. suis type 2 to adhere to macrophages. An enzyme-linked immunosorbent assay technique was standardized to simply and accurately measure the rate of bacterial attachment to phagocytic cells. Results were confirmed by plate counting. Adhesion was dependent on bacterial concentration and incubation time and was not affected by cytochalasin pretreatment of macrophages. Inhibition studies showed that the sialic acid moiety of the S. suis capsule would be, at least in part, responsible for bacterial recognition by macrophages. Serum preopsonization of bacteria increased adhesion levels. Complement would be partially implicated in the serum-enhanced binding of S. suis to cells. Adhesion varied among different S. suis type 2 isolates. However, high bacterial concentrations of several isolates were cytotoxic for cells, and these cytotoxic effects correlated with suilysin production. Indeed, hemolytic strain supernatants, as well as purified suilysin, reproduced cytotoxic effects observed with live bacteria, and these effects were inhibited by cholesterol pretreatment. Bacterial adhesion and cytotoxicity were confirmed by scanning and transmission electron microscopy. We hypothesize that attachment of bacteria to phagocytes could play an important role in the pathogenesis of S. suis infection by allowing bacterial dissemination and causing a bacteremia and/or septicemia. This interaction could also be related to the activation of the host inflammatory response observed during meningitis.
Insights
Streptococcus suis type 2 bacteria can attach to macrophages, aiding their spread in the body. This adhesion, influenced by the bacterial capsule and serum, may contribute to swine meningitis and zoonotic infections.
Area of Science:
- Veterinary Microbiology
- Immunology
- Pathogenesis
Background:
- Streptococcus suis capsular type 2 is a significant cause of swine meningitis and a zoonotic pathogen.
- Infection pathogenesis is hypothesized to involve bacteria associating with mononuclear phagocytes to invade tissues.
Purpose of the Study:
- To investigate the adherence capacity of Streptococcus suis type 2 to macrophages.
- To understand the mechanisms underlying bacterial-macrophage interaction in S. suis infections.
Main Methods:
- Standardized an enzyme-linked immunosorbent assay (ELISA) to quantify bacterial attachment to phagocytic cells.
- Confirmed adhesion rates using plate counting.
- Utilized scanning and transmission electron microscopy for visual confirmation.
- Investigated the role of sialic acid, serum opsonization, and complement in adhesion.
- Assessed bacterial cytotoxicity and its correlation with suilysin production.
Main Results:
- Bacterial adhesion to macrophages was dependent on bacterial concentration and incubation time.
- The sialic acid component of the S. suis capsule partially mediated recognition by macrophages.
- Serum preopsonization significantly increased bacterial adhesion, with complement playing a partial role.
- Cytotoxicity observed at high bacterial concentrations correlated with suilysin production, which was inhibited by cholesterol.
- Adhesion and cytotoxic effects were visualized using electron microscopy.
Conclusions:
- Bacterial attachment to phagocytes is a potential key factor in Streptococcus suis pathogenesis, facilitating dissemination and leading to bacteremia/septicemia.
- The interaction between S. suis and phagocytes may also contribute to the inflammatory responses observed in meningitis.
- Understanding these interactions is crucial for developing strategies against S. suis infections.