Iranian Journal of Parasitology 2018. 13(2):215-224.

Immunogenicity of Multi-epitope Vaccine Candidate against Toxoplasma gondii Infection in BALB/c Mice
Khalid HAJISSA, Robaiza ZAKARIA, Rapeah SUPPIAN, Zeehaida MOHAMED


Background: Toxoplasma gondii is a widely prevalent intracellular protozoan parasite which causes serious clinical and veterinary problems. Development of an effective vaccine for controlling toxoplasmosis is an extremely important aim. In the present study, the protective efficacy of recombinant multiepitope antigen (USM.TOXO1) expressing nine potential epitopes identified from SAG1, GRA2, and GRA7 of Toxoplasma gondii was evaluated in BALB/c mice.

Methods: Mice were immunized subcutaneously with three doses of USM.TOXO1 antigen (10 µg/ml). Following the immunization, the IgG antibody, IgG subclass, IFN-γ and IL-4 production were evaluated using ELISA, the study was conducted at Animal Research and Service Center (ARASC), USM Health Campus in 2016.

Results: Mice immunized with USM.TOXO1 significantly induced a mixed Th1/Th2 response polarized toward the IgG1 antibody isotype. While the cytokine analysis revealed a significant release of IFN-γ cytokines.

Conclusion: USM.TOXO1 is a potential vaccine candidate that elicits strong immunity in BALB/c mice. The proven immunogenicity of the generated antigen can serve as a premise for further use of epitope-based vaccine in the immunoprevention of human and animal toxoplasmosis.


Toxoplasma gondii, Multiepitope peptide, SAG1, GRA2, GRA7, Vaccine

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Cong H, Yuan Q, Zhao Q, Zhao L, Yin H, Zhou H, He S, Wang Z. Comparative efficacy of a multi-epitope DNA vaccine via intranasal, peroral, and intramuscular delivery against lethal Toxoplasma gondii infection in mice. Parasit Vectors. 2014; 7:145.

Dubey JP. History of the discovery of the life cycle of Toxoplasma gondii. Int J Parasitol. 2009; 39(8):877-82.

Tan TG, Mui E, Cong H et al. Identification of T. gondii epitopes, adjuvants, and host genetic factors that influence protection of mice and humans. Vaccine. 2010; 28(23):3977-89.

Zhang TE, Yin LT, Li RH, Wang HL, Meng XL, Yin GR. Protective immunity induced by peptides of AMA1, RON2 and RON4 containing T-and B-cell epitopes via an intranasal route against toxoplasmosis in mice. Parasit Vectors. 2015; 8:15.

Liu Q, Singla LD, Zhou H. Vaccines against Toxoplasma gondii: status, challenges and future directions. Hum Vaccin Immunother. 2012; 8(9):1305-8.

Mendes ÉA, Fonseca FG, Casério BM, Colina JP, Gazzinelli RT, Caetano BC: Recombinant vaccines against T. gondii: comparison between homologous and heterologous vaccination protocols using two viral vectors expressing SAG1. PloS One. 2013;8(5):e63201.

Zhang NZ, Chen J, Wang M, Petersen E, Zhu XQ. Vaccines against Toxoplasma gondii: new developments and perspectives. Expert Rev Vaccines. 2013; ;12(11):1287-99.

Garcia JL. Vaccination concepts against Toxoplasma gondii. Expert Rev Vaccines. 2009; 8(2):215-25.

Hiszczyńska-Sawicka E, Li H, Xu JB, Holec-Gąsior L, Kur J, Sedcole R, Bickerstaffe R, Stankiewicz M. Modulation of immune response to Toxoplasma gondii in sheep by immunization with a DNA vaccine encoding ROP1 antigen as a fusion protein with ovine CD154. Vet Parasitol. 2011; 183(1-2):72-8.

Wagner A, Schabussova I, Ruttkowski B et al. Prime-boost vaccination with Toxoplasma lysate antigen, but not with a mixture of recombinant protein antigens, leads to reduction of brain cyst formation in BALB/c mice. PloS One. 2014;10(5):e0126334-e0126334.

Pinzan CF, Sardinha-Silva A, Almeida F et al. Vaccination with Recombinant Microneme Proteins Confers Protection against Experimental Toxoplasmosis in Mice. PLoS One. 2015; 10(11):e0143087.

Lu G, Wang L, Zhou A et al. Epitope analysis, expression and protection of SAG5A vaccine against Toxoplasma gondii. Acta Trop. 2015; 146:66-72.

Cao A, Liu Y, Wang J, Li X, Wang S, Zhao Q, Cong H, He S, Zhou H. Toxoplasma gondii: Vaccination with a DNA vaccine encoding T-and B-cell epitopes of SAG1, GRA2, GRA7 and ROP16 elicits protection against acute toxoplasmosis in mice. Vaccine. 2015;33(48):6757-62.

Ben-Yedidia T, Arnon R. Epitope-based vaccine against influenza. Expert Rev Vaccines. 2007; 6(6):939-48.

Saha S, Raghava GP. Prediction of continuous B‐cell epitopes in an antigen using recurrent neural network. Proteins. 2006; 65(1):40-8.

Hajissa K, Zakaria R, Suppian R, Mohamed Z. Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans. Parasit Vectors. 2015; 8:315.

Dziadek B, Gatkowska J, Grzybowski M, Dziadek J, Dzitko K, Dlugonska H. The vaccine potential of three trivalent antigen-cocktails composed of recombinant ROP2, ROP4, GRA4 and SAG1 proteins against chronic toxoplasmosis in BALB/c mice. Exp Parasitol. 2012; 131(1):133-8.

Zhao G, Zhou A, Lu G et al. Identification and characterization of Toxoplasma gondii aspartic protease 1 as a novel vaccine candidate against toxoplasmosis. Parasit Vectors. 2013; 6:175.

Khan AM, Miotto O, Heiny A et al. A systematic bioinformatics approach for selection of epitope-based vaccine targets. Cell Immunol. 2006; 244(2):141-147

Cong H, Zhang M, Xin Q, Wang Z, Li Y, Zhao Q, Zhou H, He S. Compound DNA vaccine encoding SAG1/SAG3 with A2/B subunit of cholera toxin as a genetic adjuvant protects BALB/c mice against Toxoplasma gondii. Parasit Vectors. 2013;6:63.

Lim SS-Y, Othman RY: Recent Advances in Toxoplasma gondii Immunotherapeutics. Korean J Parasitol. 2014, 52(6):581-593.

Wang Y, Yin H. Research progress on surface antigen 1 (SAG1) of Toxoplasma gondii. Parasit Vectors. 2014; 7:180.

Min J, Qu D, Li C, Song X, Zhao Q, Li XA, Yang Y, Liu Q, He S, Zhou H. Enhancement of protective immune responses induced by Toxoplasma gondii dense granule antigen 7 (GRA7) against toxoplasmosis in mice using a prime-boost vaccination strategy. Vaccine. 2012; 30(38):5631-6.

Sun X, Wang Z, Li J, Wei F, Liu Q. Evaluation of an indirect ELISA using recombinant granule antigen GRA1, GRA7 and soluble antigens for serodiagnosis of Toxoplasma gondii infection in chickens. Res Vet Sci. 2015; 100:161-4.

Ching XT, Lau YL, Fong MY, Nissapatorn V. Evaluation of Toxoplasma gondii-recombinant dense granular protein (GRA2) for serodiagnosis by western blot. Parasitol Res. 2013; 112(3):1229-36.

Dziadek B, Gatkowska J, Grzybowski M, Dziadek J, Dzitko K, Dlugonska H. Toxoplasma gondii: The vaccine potential of three trivalent antigen-cocktails composed of recombinant ROP2, ROP4, GRA4 and SAG1 proteins against chronic toxoplasmosis in BALB/c mice. Exp Parasitol. 2012; 131(1):133-138.

Sayles PC, Gibson GW, Johnson LL. B cells are essential for vaccination-induced resistance to virulent Toxoplasma gondii. Infect Immun. 2000; 68(3):1026-33.

Couper KN, Roberts CW, Brombacher F, Alexander J, Johnson LL. Toxoplasma gondii-specific immunoglobulin M limits parasite dissemination by preventing host cell invasion. Infect Immun. 2005; 73(12):8060-8.

Meng M, He S, Zhao G et al. Evaluation of protective immune responses induced by DNA vaccines encoding Toxoplasma gondii surface antigen 1 (SAG1) and 14-3-3 protein in BALB/c mice. Parasit Vectors. 2012; 5:273.

Hiszczyńska-Sawicka E, Li H, Boyu Xu J et al. Induction of immune responses in sheep by vaccination with liposome-entrapped DNA complexes encoding Toxoplasma gondii MIC3 gene. Pol J Vet Sci. 2012; 15(1):3-9.

Quan J-H, Chu J-Q, Ismail HAHA, Zhou W, Jo E-K, Cha G-H, Lee Y-H. Induction of protective immune responses by a multiantigenic DNA vaccine encoding GRA7 and ROP1 of Toxoplasma gondii. Clin Vaccine Immunol. 2012,19(5):666-674.

Tao Q, Fang R, Zhang W et al. Protective immunity induced by a DNA vaccine-encoding Toxoplasma gondii microneme protein 11 against acute toxoplasmosis in BALB/c mice. Parasitol Res. 2013; 112(8):2871-7.

Dziadek B, Gatkowska J, Brzostek A, Dziadek J, Dzitko K, Dlugonska H. Toxoplasma gondii: the immunogenic and protective efficacy of recombinant ROP2 and ROP4 rhoptry proteins in murine experimental toxoplasmosis. Exp Parasitol. 2009; 123(1):81-9.

Sun X, Mei M, Zhang X et al. The extracellular matrix protein mindin as a novel adjuvant elicits stronger immune responses for rBAG1, rSRS4 and rSRS9 antigens of Toxoplasma gondii in BALB/c mice. BMC Infect Dis. 2014; 14:429.

Khorshidvand Z, Shahabi S, Mohamadzade H, Daryani A, Hazrati Tappeh K. Mixture of Alum–Naloxone and Alum–Naltrexone as a novel adjuvant elicits immune responses for Toxoplasma gondii lysate Antigen in BALB/c mice. Exp Parasitol. 2016; 162:28-34.

Garcia J, Innes EA, Katzer F. Current progress toward vaccines against Toxoplasma gondii. Vaccine Devel Ther. 2014;4:23-37.

Dziadek, B, Gatkowska, J, Grzybowski, Dziadek, J, Dzitko, K, Dlugonska H. Toxoplasma gondii: The vaccine potential of three trivalent antigen-cocktails composed of recombinant ROP2, ROP4, GRA4 and SAG1 proteins against chronic toxoplasmosis in BALB/c mice. Exp Parasitol. 2012, 131(1), 133-138.

Yin H, Zhao L, Wang T, Zhou H, He S, Cong H. A Toxoplasma gondii vaccine encoding multistage antigens in conjunction with ubiquitin confers protective immunity to BALB/c mice against parasite infection. Parasit Vectors. 2015;8(1):1-11.


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