The Cryptosporidium parvum gp60 glycoprotein expressed in the ciliate Tetrahymena thermophila is immunoreactive with sera of calves infected with Cryptosporidium oocysts

• Published in: Veterinary parasitology 2019-07, Vol.271, p.45-50
• Main Authors: Elguero, María E., Tomazic, Mariela L., Montes, María G., Florin-Christensen, Mónica, Schnittger, Leonhard, Nusblat, Alejandro D.
• Format: Article
• Language: English
• Subjects: Animals; Antigens, Protozoan - genetics; Antigens, Protozoan - immunology; Cattle; Cattle Diseases - immunology; Cattle Diseases - prevention & control; Cryptosporidiosis; Cryptosporidiosis - immunology; Cryptosporidiosis - prevention & control; Cryptosporidium parvum; Cryptosporidium parvum - genetics; Glycoproteins - genetics; Glycoproteins - immunology; Gp60; Heterologous expression; Immunoreactivity; Tetrahymena thermophila; Tetrahymena thermophila - genetics; Vaccines, Synthetic - blood; Vaccines, Synthetic - genetics
• ISSN: 0304-4017; 1873-2550
• DOI: 10.1016/j.vetpar.2019.06.008

[Display omitted] •C. parvum gp60 was efficiently expressed in the ciliate T. thermophila.•Gp60 was localized at the ciliate cell surface and oral apparatus.•Sera of calves infected with C. parvum reacted with gp60 expressed in the ciliate. Cryptosporidium parvum is a protozoan parasite of the phylum Apicomplexa responsible for cryptosporidiosis in calves, a disease that causes significant diarrhea and impairs gain of body weight, generating important production losses. As to now, no effective drugs or vaccines are available for the treatment or prevention of bovine cryptosporidiosis. Several reports suggest that development of a vaccine to prevent cryptosporidiosis is feasible, but relatively few vaccine candidates have been characterized and tested. The most prominent C. parvum antigen is gp60, an O-glycosylated mucin-like protein tethered to the parasite membrane by a glycosylphosphatidylinositol (GPI) anchor. Gp60 has been shown to be involved in essential mechanisms for the survival of C. parvum, such as recognition, adhesion to, and invasion of host cells. This work was aimed at expressing gp60 in Tetrahymena thermophila, a ciliated protozoon with numerous advantages for the heterologous expression of eukaryotic proteins, as a first approach for the development of a recombinant vaccine for bovine cryptosporidiosis. T. thermophila-expressed gp60 localized to the protozoon cell surface and oral apparatus, and partitioned into the Triton X-114 detergent phase. This indicates that the protein entered the reticuloendothelial system of the ciliate, and suggests it contains a GPI-anchor. Homogenates of gp60-expressing T. thermophila cells were recognized by sera from calves naturally infected with C. parvum demonstrating their immunoreactivity. In summary, the heterologous expression of gp60, a C. parvum-encoded GPI-anchored protein, has been successfully demonstrated in the ciliate T. thermophila.