title: Toxoplasma gondii: expression and characterization of a recombinant protein containing SAG1 and GRA2 in Pichia pastoris.
abstract: Toxoplasma gondii is an obligate intracellular protozoan which infects most species of warm-blooded animals and causes toxoplasmosis. Previous immunological and immunization studies have demonstrated the potential role of T. gondii antigens SAG1 and GRA2 as a vaccine candidate. In the present study, we have cloned, expressed, and purified a recombinant protein SAG1-GRA2 in Pichia pastoris. Results showed that P. pastoris was a robust system producing a large amount of highly purified and biological activity protein. BALB/c mice immunized with SAG1-GRA2 elicited stronger humoral and cellular responses in comparison to control groups. This immunization resulted in an enhanced Th1 immune response as measured by IgG2a antibody production and increased splenocyte IFN-gamma production, whereas no IL-4 was detected. After a lethal challenge with the highly virulent T. gondii RH strain, a prolonged survival time in SAG1-GRA2-immunized mice was observed in comparison to control groups. Our data demonstrate that SAG1-GRA2 triggered a protective response against toxoplasmosis. Therefore, SAG1-GRA2 protein might be a good candidate for the further development of a multiantigenic vaccine.
authors.affiliation: Department of Parasitology, School of Medicine, Shandong University, Jinan, 250012, People's Republic of China.
authors.forename: Huaiyu
authors.initials: H
authors.lastname: Zhou
authors.forename: Qinmin
authors.initials: Q
authors.lastname: Gu
authors.forename: Qunli
authors.initials: Q
authors.lastname: Zhao
authors.forename: Jiaqin
authors.initials: J
authors.lastname: Zhang
authors.forename: Hua
authors.initials: H
authors.lastname: Cong
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Shenyi
authors.initials: S
authors.lastname: He
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D005260:Female; D005786:Gene Expression Regulation; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010843:Pichia / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D013154:Spleen / Q000166:cytology / Q000378:metabolism; D014122:Toxoplasma / Q000235:genetics* / Q000378:metabolism*; D014124:Toxoplasmosis, Animal / Q000276:immunology
chemical_list: D000953:Antigens, Protozoan; C106990:Gra2 protein, Toxoplasma gondii; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C047168:SAG1 antigen, Toxoplasma; D015847:Interleukin-4; D007371:Interferon-gamma