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[Characteristics of exosomes secreted by Toxoplasma gondii-infected mouse dendritic DC2.4 cells]
Dongliang Li1, Shujun Yang2, Hongjuan Peng1
1Department of Pathogen Biology, School of Public Health, Southern Medical University. Guangzhou 510515, China.
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, piR-mmu-1526, piR-mmu-9082, piR-mmu-17405, and piR-mmu-25576.
Conclusions:
Toxoplasma infection causes accumulation and enrichment of exosomes secreted by DC2.4 cells with increased miRNAs and piRNAs in the exosomes.
Insights
Toxoplasma gondii infection increases exosome release from dendritic cells. These exosomes become enriched with microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs), suggesting a novel regulatory mechanism.
Area of Science:
- Immunology
- Cell Biology
- Parasitology
Background:
- Dendritic cells (DCs) are crucial for initiating immune responses.
- Exosomes are small extracellular vesicles involved in intercellular communication.
- Toxoplasma gondii is an opportunistic parasite that infects various host cells.
Purpose of the Study:
- To investigate exosome secretion changes in mouse dendritic cell line DC2.4 upon Toxoplasma gondii infection.
- To analyze the regulatory mechanisms of these exosome alterations.
Main Methods:
- Exosome isolation via ultracentrifugation from infected DC2.4 cells.
- Characterization of exosome morphology, size, and density (TEM, nanoparticle tracking).
- High-throughput sequencing for small RNA profiling in exosomes.
Main Results:
- Toxoplasma gondii infection significantly increased exosome density from DC2.4 cells.
- Increased abundance of microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs) in exosomes post-infection.
- Specific piRNAs (e.g., piR-mmu-159, piR-mmu-1526) were significantly upregulated.
Conclusions:
- Toxoplasma gondii infection induces exosome accumulation and enrichment in dendritic cells.
- Infected exosomes show increased miRNA and piRNA content, indicating altered intercellular signaling.
