Characterization of Extracellular Vesicles from Streptococcus thermophilus 065 and Their Potential to Modulate the Immune Response
2025
Ortiz Camargo, Angela Rocio | van Mastrigt, Oscar | Gouw, Joost W. | Liu, Yue | Bongers, Roger S. | van Bergenhenegouwen, Jeroen | Knol, Jan | Abee, Tjakko | Smid, Eddy J.
Bacteria can release membrane-derived nanoparticles made of lipid bilayers, so-called extracellular vesicles (EVs), which can carry diverse cargo and are important for microbe-microbe and microbe-host interactions. Here, we studied the production of EVs by Streptococcus thermophilus 065, the protein composition of the EVs, and how the produced EVs impact the immune response in vitro. Cultures of S. thermophilus grown for 6 h at 40 °C in M17 broth with 2% lactose reached high biomass yields and a high level of EVs quantified by lipophilic fluorescent dye staining. Proteome analysis of the isolated EVs revealed a high abundance of membrane-associated binding proteins of ABC transporters, ribosomal proteins, and glycolytic enzymes. In addition, phage proteins were found to be present in the EVs, which suggests a low-level expression of prophage genes during growth most likely supporting the release of EVs without causing cell lysis. The role of prophage activation was confirmed in an experiment with the addition of mitomycin C resulting in the expression of phage proteins including holin and endolysin causing a drop in culture OD and concomitant EV release. Subsequent in vitro immune assays using non-activated and activated human peripheral blood mononuclear cells (PBMCs) showed immune regulation in both cases upon exposure to S. thermophilus EVs and producer cells. This study shows the capacity of S. thermophilus EVs to act as immune modulators and opens the possibility for their use as postbiotics.
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