Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris
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 blo...
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| Format: | Article |
| Language: | English |
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Springer Verlag (Germany)
2011
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| Online Access: | http://eprints.um.edu.my/2730/1/Immunogenic_characterization_of_the_chimeric_surface_antigen_1_and_2__of_Toxoplasma_gondii_expressed_in_the_yeast_Pichia_pastoris.pdf |
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| author | Lau, Yee Ling Thiruvengadam, Girija Lee, Wei Wei Fong, Mun Yik |
| author_facet | Lau, Yee Ling Thiruvengadam, Girija Lee, Wei Wei Fong, Mun Yik |
| author_sort | Lau, Yee Ling |
| collection | UM |
| description | 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. |
| first_indexed | 2024-03-06T05:08:38Z |
| format | Article |
| id | um.eprints-2730 |
| institution | Universiti Malaya |
| language | English |
| last_indexed | 2024-03-06T05:08:38Z |
| publishDate | 2011 |
| publisher | Springer Verlag (Germany) |
| record_format | dspace |
| spelling | um.eprints-27302019-11-15T01:44:06Z http://eprints.um.edu.my/2730/ Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris Lau, Yee Ling Thiruvengadam, Girija Lee, Wei Wei Fong, Mun Yik R Medicine 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. Springer Verlag (Germany) 2011 Article PeerReviewed application/pdf en http://eprints.um.edu.my/2730/1/Immunogenic_characterization_of_the_chimeric_surface_antigen_1_and_2__of_Toxoplasma_gondii_expressed_in_the_yeast_Pichia_pastoris.pdf 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. https://doi.org/10.1007/s00436-011-2315-6 do:10.1007/s00436-011-2315-6 |
| spellingShingle | R Medicine Lau, Yee Ling Thiruvengadam, Girija Lee, Wei Wei Fong, Mun Yik Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris |
| title | Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris |
| title_full | Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris |
| title_fullStr | Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris |
| title_full_unstemmed | Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris |
| title_short | Immunogenic characterization of the chimeric surface antigen 1 and 2 (SAG1/2) of Toxoplasma gondii expressed in the yeast Pichia pastoris |
| title_sort | immunogenic characterization of the chimeric surface antigen 1 and 2 sag1 2 of toxoplasma gondii expressed in the yeast pichia pastoris |
| topic | R Medicine |
| url | http://eprints.um.edu.my/2730/1/Immunogenic_characterization_of_the_chimeric_surface_antigen_1_and_2__of_Toxoplasma_gondii_expressed_in_the_yeast_Pichia_pastoris.pdf |
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