title: Toxoplasma gondii: cloning, expression and immunoreactivity of recombinant ROP5 and ROP18 antigens.
abstract: Early diagnosis and determining the infective stage are critical for effective therapy of toxoplasmosis. Owing to the progress in biotechnology, commonly used native, non-standardized diagnostic antigens should be replaced by genetically engineered antigens. The recombinant proteins are also promising components of subunit vaccines against Toxoplasma gondii infections. A strategic biological role of rhoptry proteins (ROP) in parasitophorous vacuole biogenesis and virulence of the parasite creates a necessity for an intensive study on the serological activity and immunogenicity of newly developed recombinant ROP antigens. Our findings indicate that all generated preparations of recombinant ROP5 and ROP18 antigens, expressed in Escherichia coli bacteria, are recognized by specific antibodies produced during acute and chronic infections in inbred laboratory mice. We noticed, for the first time, that ROP5 IgM antibodies are an early and sensitive marker of T. gondii infection. The proven immunoreactivity of the obtained preparations has become a premise for a further study on their utility in routine diagnosis of human and animal toxoplasmosis as well as in the immunoprevention of T. gondii infection (as the main or supplementary component of the vaccine).
authors.affiliation: Department of Immunoparasitology, Faculty of Biology and Environmental Protection, University of Łódź, Łódź, Poland. Electronic address: marcin@biol.uni.lodz.pl.
authors.forename: Marcin M
authors.initials: MM
authors.lastname: Grzybowski
authors.affiliation: Department of Immunoparasitology, Faculty of Biology and Environmental Protection, University of Łódź, Łódź, Poland.
authors.forename: Bożena
authors.initials: B
authors.lastname: Dziadek
authors.affiliation: Institute of Medical Biology, Polish Academy of Sciences, Łódź, Poland.
authors.forename: Jarosław
authors.initials: J
authors.lastname: Dziadek
authors.affiliation: Department of Immunoparasitology, Faculty of Biology and Environmental Protection, University of Łódź, Łódź, Poland.
authors.forename: Justyna
authors.initials: J
authors.lastname: Gatkowska
authors.affiliation: Department of Immunoparasitology, Faculty of Biology and Environmental Protection, University of Łódź, Łódź, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Dzitko
authors.affiliation: Department of Immunoparasitology, Faculty of Biology and Environmental Protection, University of Łódź, Łódź, Poland.
authors.forename: Henryka
authors.initials: H
authors.lastname: Długońska
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004926:Escherichia coli; D015971:Gene Expression Regulation, Enzymologic; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000276:immunology; D015800:Protozoan Proteins; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014122:Toxoplasma / Q000201:enzymology / Q000235:genetics / Q000276:immunology*
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D017346:Protein Serine-Threonine Kinases; C556793:ROP18 protein, Toxoplasma gondii
keywords: E. coli expression system; Murine toxoplasmosis; ROP18; ROP5; Recombinant antigen; Toxoplasma gondii