Abstract: Objective To investigate the changes in the exosomes secreted by mouse dendritic cell line DC2.4 after infection with Toxoplasma gondii and to analyze the possible regulatory mechanisms underlying such changes. Methods The exosomes were extracted by ultracentrifugation from DC2.4 cells at 28 h after infection with Toxoplasma gondii. The morphology of the exosomes was examined with transmission electron microscopy, and the exosome size and density were determined using a nanoparticle tracker. High-throughput sequencing was carried out to identify the differentially expressed small RNAs in the exosomes derived from the infected cells. Results T. gondii infection resulted in a significantly increased density of exosomes secreted by DC2.4 cells. Small RNA sequencing revealed that Toxoplasma infection caused an increase in the number of miRNAs and piRNAs in the exosomes. The significantly up-regulated piRNAs after the infection included piR-mmu-159, piRmmu-1526, piR-mmu-9082, piR-mmu-17405, and piR-mmu-25576. Conclusion Toxoplasma infection causes accumulation and enrichment of exosomes secreted by DC2.4 cells with increased miRNAs and piRNAs in the exosomes.