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Monocytes are among the major myeloid cells that respond to  Toxoplasma , a ubiquitous foodborne that infects ≥ 1 billion people worldwide, in human peripheral blood. As such, a molecular understanding of human monocyte- Toxoplasma  interactions can expedite the development of novel human toxoplasmosis control strategies. Current molecular studies on monocyte- Toxoplasma  interactions are based on average cell or parasite responses across bulk cell populations. Although informative, population-level averages of monocyte responses to  Toxoplasma  have sometimes produced contradictory results, such as whether CCL2 or IL12 define effective monocyte responses to the parasite. Here, we used single-cell dual RNA sequencing (scDual-Seq) to comprehensively define, for the first time, the monocyte and parasite transcriptional responses that underpin human monocyte- Toxoplasma  encounters at the single cell level. We report extreme transcriptional variability between individual monocytes. Furthermore, we report that  Toxoplasma -exposed and unexposed monocytes are transcriptionally distinguished by a reactive subset of CD14 + CD16 -  monocytes. Functional cytokine assays on sorted monocyte populations show that the infection-distinguishing monocytes secrete high levels of chemokines, such as CCL2 and CXCL5. These findings uncover the  Toxoplasma -induced monocyte transcriptional heterogeneity and shed new light on the cell populations that largely define cytokine and chemokine secretion in human monocytes exposed to  Toxoplasma .

Single-cell RNA-seq reveals CD16- monocytes as key regulators of human monocyte transcriptional response to Toxoplasma