A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.
Eloise J LockyerFrancesca TorelliSimon ButterworthOk-Ryul SongSteven HowellAnne WestonPhilip EastMoritz TreeckPublished in: PLoS biology (2023)
Toxoplasma gondii secretes protein effectors to subvert the human immune system sufficiently to establish a chronic infection. Relative to murine infections, little is known about which parasite effectors disarm human immune responses. Here, we used targeted CRISPR screening to identify secreted protein effectors required for parasite survival in IFNγ-activated human cells. Independent screens were carried out using 2 Toxoplasma strains that differ in virulence in mice, leading to the identification of effectors required for survival in IFNγ-activated human cells. We identify the secreted protein GRA57 and 2 other proteins, GRA70 and GRA71, that together form a complex which enhances the ability of parasites to persist in IFNγ-activated human foreskin fibroblasts (HFFs). Components of the protein machinery required for export of Toxoplasma proteins into the host cell were also found to be important for parasite resistance to IFNγ in human cells, but these export components function independently of the identified protein complex. Host-mediated ubiquitination of the parasite vacuole has previously been associated with increased parasite clearance from human cells, but we find that vacuoles from GRA57, GRA70, and GRA71 knockout strains are surprisingly less ubiquitinated by the host cell. We hypothesise that this is likely a secondary consequence of deletion of the complex, unlinked to the IFNγ resistance mediated by these effectors.
Keyphrases
- toxoplasma gondii
- immune response
- plasmodium falciparum
- dendritic cells
- endothelial cells
- protein protein
- escherichia coli
- trypanosoma cruzi
- amino acid
- induced pluripotent stem cells
- single cell
- type iii
- cell therapy
- binding protein
- genome wide
- pseudomonas aeruginosa
- staphylococcus aureus
- adipose tissue
- pluripotent stem cells
- insulin resistance
- bone marrow
- dna methylation
- genome editing
- candida albicans