Characterizing Cystoisospora canis as a Model of Apicomplexan Tissue Cyst Formation and Reactivation
Houk-Miles, Alice Elizabeth
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Cystoisospora canis is an Apicomplexan parasite of the small intestine of dogs. C. canis produces monozoic tissue cysts (MZT) that are similar to the polyzoic tissue cysts (PZT) of Toxoplasma gondii, a parasite of medical and veterinary importance, which can reactivate and cause toxoplasmic encephalitis. We hypothesized that C. canis is similar biologically and genetically enough to T. gondii to be a novel model for studying tissue cyst biology. We examined the pathogenesis of C. canis in beagles and quantified the oocysts shed. We found this isolate had similar infection patterns to other C. canis isolates studied. We were able to superinfect beagles that came with natural infections of Cystoisospora ohioensis-like oocysts indicating that little protection against C. canis infection occurred in these beagles. The C. canis oocysts collected were purified and used for future studies. We demonstrated in vitro that C. canis could infect 8 mammalian cell lines and produce MZT. The MZT were able to persist in cell culture for at least 60 days. We were able to induce reactivation of MZT treated with bile-trypsin solution. In molecular studies, we characterized C. canis genetically using ITS1 and CO1 to build phylogenetic trees and found C. canis was most similar to C. ohioensis-like with ITS1 and more similar to T. gondii than any other coccidia using ITS1 and CO1. We identified genes and proteins involved with virulence, cyst wall structure, and immune evasion of T. gondii and examined the DNA of C. canis for orthologs. C. canis had orthologs with 8 of 20 T. gondii genes examined. Monoclonal and polyclonal antibody and lectin studies demonstrated similar tissue cyst wall proteins on C. canis MZT and T. gondii PZT. Our findings in vitro and using genetic characterization of C. canis indicated the presence of similar genes and proteins, and its close phylogenetic location with T. gondii demonstrate that C. canis may serve as a model to examine tissue cyst biology. The system we described provides a simple model to produce tissue cysts and to study host factors that cause reactivation of tissue cysts.
- Doctoral Dissertations