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Sci Rep. 2017 Jun 19;7(1):3768. doi: 10.1038/s41598-017-03978-1.

New molecular tools in Neospora caninum for studying apicomplexan parasite proteins.

Author information

1
Laboratory of Immunoparasitology "Dr. Mário Endsfeldz Camargo", Institute of Biomedical Sciences, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais, Brazil.
2
Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, California, USA.
3
Department of Biological Chemistry and Institute of Genomics and Proteomics, University of California, Los Angeles, Los Angeles, California, USA.
4
Laboratory of Immunoparasitology "Dr. Mário Endsfeldz Camargo", Institute of Biomedical Sciences, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais, Brazil. tiago.mineo@ufu.br.
5
Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, California, USA. pbradley@ucla.edu.
6
Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California, USA. pbradley@ucla.edu.

Abstract

The development of molecular genetics has greatly enhanced the study of the biology and pathology associated with parasites of the phylum Apicomplexa. We have established a system specifically designed for Neospora caninum, and used this system as a heterologous platform for the expression of foreign genes. Plasmid constructs containing fluorescent proteins or targeted genes of Toxoplasma gondii, driven by N. caninum promoters, have yielded robust expression and correct trafficking of target gene products as assessed by immunofluorescence assays and Western blot analyses. Using this approach, we here demonstrated that N. caninum expressing T. gondii's GRA15 and ROP16 kinase are biologically active and induced immunological phenotypes consistent with T. gondii strains. N. caninum expressing TgGRA15 differentially disturbed the NF-κB pathway, inducing an increased IL-12 production. On the other hand, N. caninum expressing TgROP16 induced host STAT3 phosphorylation and consequent reduction of IL-12 synthesis. These results indicate that heterologous gene expression in N. caninum is a useful tool for the study of specific gene functions and may allow the identification of antigenic targets responsible for the phenotypic differences observed between these two closely related apicomplexan parasites. Additionally, these observations may prove to be useful for the development of vaccine protocols to control toxoplasmosis and/or neosporosis.

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