Cytokine array analysis of mediators produced by human macrophages stimulated with Trichomonas tenax

Trichomonas tenax is one of two oral protozoans that colonize dental plaque, periodontal pockets, and the surfaces of the teeth and gingiva in persons with poor oral hygiene. Its prevalence varies widely in the human population, ranging from a high of 56% in Chile to a low of 3% in Kenya, with a global pooled prevalence of 17% (Eslahi et al., 2021). The second broadly disseminated oral protozoan is Entamoeba gingivalis, which has a pooled prevalence of 37% (Badri et al., 2021). Once considered a commensal or saprophytic organism, T. tenax is gaining recognition as a potential oral pathogen. Benabdelkader et al. (2019) showed that clinical isolates of T. tenax are clustered into three groups, and that strains from two groups are closely associated with severe periodontitis. Differences in virulence factors between the groups might explain why some are more closely aligned with periodontitis than others, e.g., clinical isolates differ in the tissue-damaging proteases that they produce (El Sibaei et al., 2012). Further, certain isolates of T. tenax might affect oral health by inducing formation of apoptotic bodies in mammalian cells, a mechanism that has been demonstrated for the protozoan in vitro (Ribeiro et al., 2015), or by altering the composition of the oral microbiome in favor of pathogenic microorganisms (Marty et al., 2017).

The association between T. tenax oral colonization and periodontitis warrants further investigation. Advanced periodontitis elicits a proinflammatory cytokine response that contributes locally to alveolar bone loss and systemically to inflammatory conditions like arthritis and coronary heart disease (Chen et al., 2018). A previous study in our laboratory showed that at low protozoan-to-macrophage ratios (no more than 1 protozoan per 5 macrophages), T. tenax elicited only modest interleukin-8 production from the human macrophage cell line THP-1, but did not elicit IL-1β, IL-10, or tumor necrosis factor α (TNFα) (Govro and Stuart, 2016). In the current study, THP-1 macrophages were cultured with T. tenax or the urogenital pathogen T. vaginalis at a ratio of 5 protozoans to one macrophage to mimic a more advanced infection, and the breadth of the cytokine panel was increased from 4 to 36 cytokines. Here we report the results of cytokine array analysis performed on conditioned media harvested from THP-1 macrophages stimulated with live or sonicated trichomonads.

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