Identification of Cytauxzoon felis antigens via protein microarray and assessment of expression library immunization against cytauxzoonosis

通过蛋白质微阵列鉴定猫胞囊虫抗原及表达文库对猫胞囊虫病免疫效果的评估

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作者:Megan E Schreeg, Henry S Marr, Jaime L Tarigo, Meredith K Sherrill, Hilton K Outi, Elizabeth H Scholl, David M Bird, Adam Vigil, Chris Hung, Rie Nakajima, Li Liang, Angela Trieu, Denise L Doolan, Jennifer E Thomas, Michael G Levy, Mason V Reichard, Philip L Felgner, Leah A Cohn, Adam J Birkenheuer

Background

Cytauxzoonosis is a disease of felids in North America caused by the tick-transmitted apicomplexan parasite Cytauxzoon felis. Cytauxzoonosis is particularly virulent for domestic cats, but no vaccine currently exists. The parasite cannot be cultivated in vitro, presenting a significant limitation for vaccine development.

Conclusions

Protein microarray facilitated high-throughput identification of novel antigens, substantially increasing the pool of characterized C. felis antigens. These antigens should be considered for development of C. felis vaccines, diagnostics, and therapeutics.

Methods

Recent sequencing of the C. felis genome has identified over 4300 putative protein-encoding genes. From this pool we constructed a protein microarray containing 673 putative C. felis proteins. This microarray was probed with sera from C. felis-infected and naïve cats to identify differentially reactive antigens which were incorporated into two expression library vaccines, one polyvalent and one monovalent. We assessed the efficacy of these vaccines to prevent of infection and/or disease in a tick-challenge model.

Results

Probing of the protein microarray resulted in identification of 30 differentially reactive C. felis antigens that were incorporated into the two expression library vaccines. However, expression library immunization failed to prevent infection or disease in cats challenged with C. felis. Conclusions: Protein microarray facilitated high-throughput identification of novel antigens, substantially increasing the pool of characterized C. felis antigens. These antigens should be considered for development of C. felis vaccines, diagnostics, and therapeutics.

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