Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris

Lau, Yee Ling and Thiruvengadam, Girija and Lee, Wei Wei and Fong, Mun Yik (2011) Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris. Parasitology Research, 109 (3). pp. 871-878. ISSN 0932-0113, DOI do:10.1007/s00436-011-2315-6.

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Abstract

In this study, we successfully expressed a chimerical surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii in Pichia pastoris. Eighty human serum samples, including 60 from confirmed cases of toxoplasmosis, were tested against the purified recombinant SAG1/2 in Western blots. Results of Western blots targeted at Toxoplasma IgG and IgM showed that the recombinant SAG1/2 reacted with all sera from the toxoplasmosis cases but none with the Toxoplasma-negative serum samples. These results showed that the P. pastoris-derived recombinant SAG1/2 was sensitive and specific and suitable for use as antigen for detecting anti-Toxoplasma antibodies. To further investigate the immunological characteristic of the recombinant protein, the recombinant SAG1/2 was injected subcutaneously into BALB/c mice, and their serum was tested against total protein lysate of T. gondii. Mice immunized with the recombinant SAG1/2 reacted specifically with the native SAG1 and SAG2 of T. gondii. Significant proliferation of splenocytes stimulated with tachyzoite total protein lysate was observed in vaccinated BALB/c mice but not in those from negative control mice. Specific production of IFN-gamma, the Th1-type cytokines, was also found in stimulated splenocytes from vaccinated mice. These results show that the chimeric protein recombinant SAG1/2 can elicit a Th1-associated protection against T. gondii infections in mice. Finally, vaccinated mice were significantly protected against lethal challenge with live T. gondii RH strain tachyzoites (P < 0.005), and their survival time increased significantly compared to the negative control.

Item Type:	Article
Funders:	UNSPECIFIED
Additional Information:	Lau, Yee Ling Thiruvengadam, Girija Lee, Wei Wei Fong, Mun Yik
Uncontrolled Keywords:	Toxoplasmosis; Recombinant Antigen; Toxocariasis; SAG2 Gene; Total Protein Lysate
Subjects:	R Medicine
Divisions:	Faculty of Medicine
Depositing User:	Prof. Dr. Mun Yik Fong
Date Deposited:	22 Feb 2012 07:41
Last Modified:	15 Nov 2019 01:44
URI:	http://eprints.um.edu.my/id/eprint/2730

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