Immunogenicity of Chitosan, Montanide and Aluminum hydroxide as an adjuvants in Toxoplasma gondii experimental vaccine in sheep

Document Type : Full Research Paper

Authors

1 Department of Veterinary Medicine, Faculty of Veterinary Medicine, Kazerun Branch, Islamic Azad University, Kazerun, Iran

2 Razi Serum and Vaccine Research Institute, Agricultural Research, Education and Extension Organization, Shiraz, Iran.

3 Department of Parasitology, Faculty of Veterinary Medicine, Kazerun Branch, Islamic Azad University, Kazerun, Iran

4 Department of veterinary medicine, Faculty of veterinary medicine, Kazerun Branch, Islamic Azad university, Kazerun, Iran

5 Razi vaccine and serum research institute, Shiraz Branch, Agricultural Research, Education and Extension Organization (AREEO), Shiraz, Iran

Abstract

Toxoplasma gondii is a zoonoses protozoan parasite that causes toxoplasmosis. This parasite causes severe economic losses in the sheep industry. The aim of this study was to evaluate the immunogenicity of the live attenuated vaccine with different adjuvants in sheep. 20 two-month-old pregnant Iranian Qashqaeian serum-negative sheep for Toxoplasma gondii were randomly divided into five groups of four. Group one injected with media and served as control. Three experimental groups were immunized subcutaneously with 1 ml of 25 ×106 inactivate tachyzoites formulated with Montanide, alum, and chitosan, respectively. Last group was inoculated with live attenuated tachyzoite with the same dose. Immunization was performed once. After 21 days, blood samples were collected for ELISA test to evaluate the humoral and cellular immune response. The highest antibody titer was obtained in the group immunized with montanide adjuvant, which was significantly different from other groups (P <0.05). However, the highest cellular immunity according to gamma interferon measurements was related to live attenuated vaccine which was significantly different from other groups, followed by chitosan or aluminum hydroxide adjuvant. It should b notice that all immunized groups had a significant difference with the control group (P <0.05). According to the successful immune response in the live attenuated vaccine, we concluded that this strain can be used for immunization of sheep as the target host and use in higher numbers of sheeps.

Keywords


1. Abbasifar, A., Namavari, M., & Rezayian, A. (2017). Evaluation of Razi attenuated variety of Toxoplasma gondii in Balb/c mice. Veterinary Researches & Biological Products, 30(4), 107-113. (In persian)
2. Buxton, D. (1998). Protozoan infections (Toxoplasma gondii, Neospora caninum and Sarcocystis spp.) in sheep and goats: recent advances. Veterinary research, 29(3-4), 289-310.
3. Buxton, D., & Innes, E. A. (1995). A commercial vaccine for ovine toxoplasmosis. Parasitology, 110(S1), S11-S16.
4. Dunay, I. R., Gajurel, K., Dhakal, R., Liesenfeld, O., & Montoya, J. G. (2018). Treatment of toxoplasmosis: historical perspective, animal models, and current clinical practice. Clinical microbiology reviews, 31(4), e00057-17.
5. Fulton, J. D., & Sutton, R. N. P. (1962). Pure suspension of Toxoplasma gondii for use in complement-fixation test. Immunology, 5(5), 621-6.
6. Gazzinelli, R. T., Wysocka, M., Hayashi, S., Denkers, E. Y., Hieny, S., Caspar, P., ... & Sher, A. (1994). Parasite-induced IL-12 stimulates early IFN-gamma synthesis and resistance during acute infection with Toxoplasma gondii. The Journal of Immunology, 153(6), 2533-2543.
7. Guo, J., Sun, X., Yin, H., Wang, T., Li, Y., Zhou, C., ... & Cong, H. (2018). Chitosan microsphere used as an effective system to deliver a linked antigenic peptides vaccine protect mice against acute and chronic toxoplasmosis. Frontiers in cellular and infection microbiology, 8, 163.
8. Ibrahim, E. E. S., Gamal, W. M., Hassan, A. I., Mahdy, S. E. D., Hegazy, A. Z., & Abdel-Atty, M. M. (2015). Comparative study on the immunopotentiator effect of ISA 201, ISA 61, ISA 50, ISA 206 used in trivalent foot and mouth disease vaccine. Veterinary World, 8(10), 1189.
9. Ismael, A. B., Sekkai, D., Collin, C., Bout, D., & Mévélec, M. N. (2003). The MIC3 gene of Toxoplasma gondii is a novel potent vaccine candidate against toxoplasmosis. Infection and immunity, 71(11), 6222-6228.
10. Jang, S. I., Lillehoj, H. S., Lee, S. H., Lee, K. W., Lillehoj, E. P., Bertrand, F., ... & Deville, S. (2011). Montanide™ ISA 71 VG adjuvant enhances antibody and cell-mediated immune responses to profilin subunit antigen vaccination and promotes protection against Eimeria acervulina and Eimeria tenella. Experimental parasitology, 127(1), 178-183.
11. Kang, H. J., Kim, M. J., Chu, K. B., Lee, S. H., Moon, E. K., & Quan, F. S. (2021). Passive Immunity and Antibody Response Induced by Toxoplasma gondii VLP Immunization. Vaccines, 9(5), 425.
12. Meshkini, E., Aminpour, A., Tappeh, K. H., Seyyedi, S., & Shokri, M. (2021). Evaluation of Adjuvant Effectiveness of Alum-Propranolol Mixture on the Immunogenicity of Excreted/Secreted Antigens of Toxoplasma gondii RH Strain. Advanced Pharmaceutical Bulletin, 11(3), 570.
13. Petersen, E., Nielsen, H. V., Christiansen, L., & Spenter, J. (1998). Immunization with E. coli produced recombinant T. gondii SAG1 with alum as adjuvant protect mice against lethal infection with Toxoplasma gondii. Vaccine, 16(13), 1283-1289.
14. Rémy, V., Zöllner, Y., & Heckmann, U. (2015). Vaccination: the cornerstone of an efficient healthcare system. Journal of market access & health policy, 3(1), 27041.
15. Robert-Gangneux, F., & Dardé, M. L. (2012). Epidemiology of and diagnostic strategies for toxoplasmosis. Clinical microbiology reviews, 25(2), 264-296.
16. Şahar, E. A., Can, H., İz, S. G., Döşkaya, A. D., Kalantari-Dehaghi, M., Deveci, R., ... & Döşkaya, M. (2020). Development of a hexavalent recombinant protein vaccine adjuvanted with Montanide ISA 50 V and determination of its protective efficacy against acute toxoplasmosis. BMC Infectious Diseases, 20(1), 1-16.
17. Sánchez-Sánchez, R., Ferre, I., Regidor-Cerrillo, J., Gutiérrez-Expósito, D., Ferrer, L. M., Arteche-Villasol, N., ... & Benavides, J. (2019). Virulence in mice of a Toxoplasma gondii type II isolate does not correlate with the outcome of experimental infection in pregnant sheep. Frontiers in cellular and infection microbiology, 8, 436.
18. Setasimy, A., & Namavari, M. (2016). Use of chicken embryonated eggs for evaluating the virulence of Toxoplasma gondii. Journal of Parasitic Diseases, 40(4), 1223-1225.
19. Shokri, M., Roohvand, F., Alimohammadian, M. H., Ebrahimirad, M., & Ajdary, S. (2016). Comparing Montanide ISA 720 and 50-V2 adjuvants formulated with LmSTI1 protein of Leishmania major indicated the potential cytokine patterns for induction of protective immune responses in BALB/c mice. Molecular immunology, 76, 108-115.
20. Sun, B., Yu, S., Zhao, D., Guo, S., Wang, X., & Zhao, K. (2018). Polysaccharides as vaccine adjuvants. Vaccine, 36(35), 5226-5234.
21. Wang, J. L., Zhang, N. Z., Li, T. T., He, J. J., Elsheikha, H. M., & Zhu, X. Q. (2019). Advances in the development of anti-Toxoplasma gondii vaccines: challenges, opportunities, and perspectives. Trends in parasitology, 35(3), 239-253.
22. Yang, J., Yang, C., Qian, J., Li, F., Zhao, J., & Fang, R. (2020). Toxoplasma gondii α‐amylase deletion mutant is a promising vaccine against acute and chronic toxoplasmosis. Microbial biotechnology, 13(6), 2057-2069.
23. Yap, G. S., & Sher, A. (1999). Effector cells of both nonhemopoietic and hemopoietic origin are required for interferon (IFN)-γ–and tumor necrosis factor (TNF)-α–dependent host resistance to the intracellular pathogen, Toxoplasma gondii. The Journal of experimental medicine, 189(7), 1083-1092.
24. Zhang, N. Z., Wang, M., Xu, Y., Petersen, E., & Zhu, X. Q. (2015). Recent advances in developing vaccines against Toxoplasma gondii: an update. Expert Review of Vaccines, 14(12), 1609-1621.
25. Zhuo, X., Du, K., Ding, H., Lou, D., Zheng, B., & Lu, S. (2021). A Carbamoyl Phosphate Synthetase II (CPSII) Deletion Mutant of Toxoplasma gondii Induces Partial Protective Immunity in Mice. Frontiers in microbiology, 3549.