Abstract
Toxoplasma gondii (T. gondii) can cause zoonotic toxoplasmosis worldwide. Neutrophil extracellular traps (NETs) have been known as a novel effector mechanism against T. gondii infection in the innate system of humans, cats, and sheep. Dogs are the intermediate host of T. gondii, in which the use of NETs against T. gondii infection remains unclear. Thus, this study aims to examine the effects of T. gondii on NETs release in dogs, and to further investigate the mechanism involved in the process. T. gondii-triggered NETs were analyzed by scanning electron microscopy (SEM) and fluorescence confocal microscopy, and the mechanism of T. gondii-triggered NETs release was determined by using inhibitors and a fluorometric reader. The results showed that T. gondii tachyzoites significantly triggered NETs-like structures, which consisted of DNA decorated with neutrophil elastase (NE) and myeloperoxidase (MPO). Further investigations revealed that reactive oxygen species (ROS)-, NADPH oxidase-, Rac 1- or p38 mitogen-activated protein kinase (MAPK)-signaling pathways were relevant to T. gondii tachyzoites-triggered NETs release. Moreover, zymosan-triggered NETs release was strikingly degraded by T. gondii tachyzoites treatment, indicating that T. gondii may escape from the NETs-based capture strategy. Taken together, promoting NETs release is suggested to limit motility and evade infection of T. gondii in dogs.
Highlights
Toxoplasma gondii is considered as one of the most prevalent zoonotic parasites worldwide (Tenter et al, 2000; Dubey and Jones, 2008; Dubey and Dubey, 2010; Lopes et al, 2014; Pan et al, 2017)
If T. gondii tachyzoites-triggered neutrophils extracellular trap (NET) have similar characteristics to the typical NETs-structures, the constituent proteins in T. gondii tachyzoites-triggered NETs-like network structures were analyzed. These results showed that T. gondii tachyzoites significantly triggered NETs-like structures, and these structures consisted of DNA
Further experiments revealed that T. gondii tachyzoitestriggered NETs release within 60 min which was in a timeindependent manner (P = 0.002, n = 5), and it was of great interest that the increasing of NETs release significantly decreased at the time point of 90 min (P = 0.034, n = 5) and 120 min (P = 0.925, n = 5; Figure 3B)
Summary
Toxoplasma gondii is considered as one of the most prevalent zoonotic parasites worldwide (Tenter et al, 2000; Dubey and Jones, 2008; Dubey and Dubey, 2010; Lopes et al, 2014; Pan et al, 2017). Dogs are the intermediate hosts, and multiple studies have demonstrated the prevalence of T. gondii in dogs in a variety of countries or areas worldwide (Jiang et al, 2015; Zhang et al, 2015; Dubey et al, 2016; Rengifo-Herrera et al, 2017; Zarra-Nezhad et al, 2017). These serological surveys and diagnosis analysis of dogs infected T. gondii have been reported, the interactions between the host dog and the parasite T. gondii remain not fully understood.
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