Compound DNA vaccine encoding SAG1/ SAG3 with A2/B subunit of cholera toxin as a genetic adjuvant protects BALB/c mice against Toxoplasma gondii

Particle and Fibre Toxicology

Shenyi He³  Huaiyu Zhou³  Qunli Zhao³  Ying Li³  Zhiyu Wang¹  Qing Xin²  Min Zhang³  Hua Cong³ 

[1] School of Public Health, Shandong University, No44 wenhuaxi Road, Jinan, Shandong, 250012, P. R. China;School hospital of Shandong University, Jinan, Shandong, 250012, P. R. China;Department of human parasitology, Shandong University School of Medicine, No44 wenhuaxi Road, Jinan, Shandong, 250012, P. R. China

关键词: DNA vaccination;    CTXA₂/B;    SAG3;    SAG1;    Surface antigen;    Toxoplasma gondii;   

Others  :  1228041
DOI  :  10.1186/1756-3305-6-63

 received in 2013-01-11, accepted in 2013-03-11,  发布年份 2013

【 摘 要 】

Background

Intracellular parasites, such as T. gondii, present a plurality of antigens because of the complexity of its life cycle. Compound DNA vaccines bring a new approach and hope for the treatment of toxoplasmosis. In this study, a DNA vaccine encoding two major surface antigens SAG1, SAG3 from T. gondii, with A₂/B subunit of cholera toxin as a genetic adjuvant was constructed.

Methods

BALB/c mice were immunized intramuscularly with PBS, pcDNA3.1, pSAG1, pSAG1/SAG3 and pSAG1/SAG3-CTXA₂/B three times separately. Immunized mice were tested for IgG antibody and IFN-γ and IL-4 production by ELISA. The proliferation of T cells was measured by DNA synthesis assay and the lymphocyte subsets of spleen cells by flow cytometry. All the immunized mice were challenged with 10³ highly virulent RH tachyzoites of Toxoplasma gondii intraperitoneally and the survival times were recorded.

Results

An enhanced production of IgG antibodies, antigen-specific lymphocyte proliferation and IFN-γ production from splenic cells were induced in mice immunized with pSAG1/SAG3 compared to mice immunized with pSAG1 (P<0.05). Introduction of CTXA₂/B further enhanced the Th1 cell-mediated immunity with higher levels of IFN-γ, lymphocyte proliferation activity and percentage of CD8⁺ T-cells. When challenged with lethal doses of T. gondii (1×10³), all control mice (PBS and empty plasmid group) died within 6 days. Mice immunized with pSAG1 died within 8 days. While 20% and 40% survival rate were achieved from mice immunized with pSAG1/SAG3 and pSAG1/SAG3-CTXA₂/B.

Conclusions

This study indicates the compound DNA vaccine encoding T. gondii antigens SAG1, SAG3 with CTXA₂/B gene was a promising DNA vaccine candidate against toxoplasmosis, which could effectively enhance the humoral and cellular immune response and prolong survival time in vaccinated mice.

【 授权许可】

   
2013 Cong et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Zhou P, Chen Z, Li HL, Zheng H, He S, Lin R, Zhu XQ: Toxoplasma gondii infection in humans in China. Parasit Vectors 2011, 4:165. BioMed Central Full Text
• [2]McLeod R, Boyer K, Roizen N: The child with congenital toxoplasmosis. Curr Clin Top Infect Dis 2000, 20:189-208.
• [3]Vittecoq M, Elguero E, Lafferty KD, Roche B, Brodeur J, Gauthier-Clerc M, Missé D, Thomas F: Brain cancer mortality rates increase with Toxoplasma gondii seroprevalence in France. Infect Genet Evol 2012, 12(2):496-498.
• [4]Walle F, Kebede N, Tsegaye A, Kassa T: Seroprevalence and risk factors for Toxoplasmosis in HIV infected and non-infected individuals in Bahir Dar. Northwest Ethiopia. Parasit Vectors 2013, 6:15. BioMed Central Full Text
• [5]Buxton D, Innes EA: A commercial vaccine for ovine toxoplasmosis. Parasitol 1995, 110:11-16.
• [6]Lunden A, Lovgren K, Ugglaand A, Araujo FG: Immune response and resistance to T. gondii in mice immunized with antigens of the parasite incorporated into immunostimulation complex. Infect Immun 1993, 61(6):2639.
• [7]Angus CW: Nucleic acid vaccination against Toxoplasma gondii in mice. J Eukaryot Microbiol 1996, 43(5):117-118.
• [8]Fachado A, Rodriguez A, Angel SO: Protective effect of a naked DNA vaccine cocktail against lethal toxoplasmosis in mice. Vaccine 2003, 21(13–14):1327-1335.
• [9]Sun XM, Zou J, Elashram Saeed AA, Yan WC, Liu XY, Suo X, Wang H, Chen QJ: DNA vaccination with a gene encoding Toxoplasma gondii GRA6 induces partial protection against toxoplasmosis in BALB/c mice. Parasit Vectors 2011, 4:213. BioMed Central Full Text
• [10]Calvin J, Cleo Y, Lee F, Michael EG: The SAS superfamily of Toxoplasma surface proteins. Int J Parasitol 2004, 34:285-296.
• [11]Meng M, He S, Zhao G, Bai Y, Zhou H, Cong H, Lu G, Zhao Q, Zhu XQ: 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. BioMed Central Full Text
• [12]Mévéle MN, Bout D, Desolme B: Evaluation of protective effect of DNA vaccination with genes encoding antigens GRA4 and SAG1 associated with GM-CSF plasmid, against acute, chronical and congenital toxoplasmosis in mice. Vaccine 2005, 23(36):4489-4499.
• [13]Rabie MM, Fumie A, Mei C: Induction of protective immunity by DNA vaccination with Toxoplasma gondii HSP70, HSP30 and SAG1 genes. Vaccine 2003, 21:2852-2861.
• [14]Cesbron-Delauw MF, Tomavo S, Beauchamps P, Fourmaux MP, Camus D, Capron A, Dubremetz JF: Similarities between the primary structures of two distinct major surface proteins of Toxoplasma gondii. J Biol Chem 1994, 269:16217-16222.
• [15]Tomavo S: The major surface proteins of Toxoplasma gondii: structures and functions. Curr Top Microbiol Immunol 1996, 219:45-54.
• [16]Lekutis C, Ferguson DJ, Grigg ME, Camps M, Boothroyd JC: Surface antigens of Toxoplasma gondii: variations on a theme. Int J Parasitol 2001, 31(12):1285-1292.
• [17]Chuang SC, Ko JC, Chen CP, Du JT, Yang CD: Induction of long-lasting protective immunity against Toxoplasma gondii in BALB/c mice by recombinant surface antigen 1 protein encapsulated in poly (lactide-co-glycolide) microparticles. Parasit Vectors 2013, 6:34. BioMed Central Full Text
• [18]Jacquet A, Coulon L, De Neve J, Daminet V, Haumont M, Garcia L, Bollen A, Jurado M, Biemans R: The surface antigen SAG3 mediates the attachment of Toxoplasma gondii to cell-surface proteoglycans. Mol Biochem Parasitol 2001, 116:35-44.
• [19]Dzierszinski F, Mortuaire M, Cesbron-Delauw MF, Tomavo S: Targeted disruption of the glycosylphosphatidylinositol-anchored surface antigen SAG3 gene in Toxoplasma gondii decreases host cell adhesion and drastically reduces virulence in mice. Mol Microbiol 2000, 37:574-582.
• [20]Angus CW, Klivington-Evans D, Dubey JP: Immunization with a DNA plasmid encoding the SAG1 (P30) protein of Toxoplasma gondii is immunogenic and protective in rodents. Infect Dis 2000, 181:317-324.
• [21]Nielsen HV, Lauemoller SL, Christiansen L, Buus S, Fomsgaard A, Petersen E: Complete protection against lethal Toxoplasma gondii infection in mice immunized with a plasmid encoding the SAG1 gene. Infect Immun 1999, 67(12):6358-6363.
• [22]Couper KN, Nielsen HV, Petersen E, Roberts F, Roberts CW, Alexander J: DNA vaccination with the immunodominant tachyzoite surface antigen (SAG-1) protects against adult acquired Toxoplasma gondii infection but does not prevent maternofoetal transmission. Vaccine 2003, 21(21–22):2813-2820.
• [23]Lee YH, Shin DW, Lee JH, Nam HW, Ahn MH: Vaccination against murine Toxoplasmosis using recombinant Toxoplasma gondii SAG3 antigen alone or in combination with Quil A. Yonsei Med J 2007, 48(3):396-404.
• [24]Lycke N: The mechanism of cholera toxin adjuvanticity. Res Immunol 1997, 148(8–9):504.
• [25]Eriksson K, Fredriksson M, Nordstrom I, Holmgren J: Cholera toxin and its B subunit promote dendritic cell vaccination with different influences on Th1 and Th2 development. Infect Immun 2003, 71(4):1740-1747.
• [26]Gazzinelli RT, Hakim FT, Hieny S, Shearer GM, Sher A: Synergistic role of CD4+ and CD8+ T lymphocytes in IFN-gamma production and protective immunity induced by an attenuated Toxoplasma gondii vaccine. J Immunol 1991, 146:286-292.
• [27]Jongert E, Lemiere A, Van Ginderachter J, De Craeye S, Huygen K, D’Souza S: Functional characterization of in vivo effector CD4(+) and CD8(+) T cell responses in acute Toxoplasmosis: an interplay of IFN-gamma and cytolytic T cells. Vaccine 2010, 28:2556-2564.
• [28]Wang X, Claflin J, Kang H, Suzuki Y: Importance of CD8(+)Vbeta8(+) T cells in IFN-gamma-mediated prevention of toxoplasmic encephalitis in genetically resistant BALB/c mice. J Interferon Cytokine Res 2005, 25(6):338-344.
• [29]Rezende-Oliveira K, Silva NM, Mineo JR, Rodrigues JV: Cytokines and chemokines production by mononuclear cells from parturient women after stimulation with live Toxoplasma gondii. Placenta 2012, 33(9):682-687.
• [30]Zou J, Huang XX, Yin GW, Ding Y, Liu XY, Wang H, Chen QJ, Suo X: Evaluation of Toxoplasma gondii as a live vaccine vector in susceptible and resistant hosts. Parasit Vectors 2011, 4:168. BioMed Central Full Text
• [31]Wu XN, Lin J, Lin X, Chen J, Chen ZL, Lin JY: Multicomponent DNA vaccine-encoding Toxoplasma gondii GRA1 and SAG1 primes: anti-Toxoplasma immune response in mice. Parasitol Res 2012, 111(5):2001-2009.
• [32]Wang JL, Liu XL, Jia B, Lu H, Peng S, Piao X, Hou N, Cai P, Yin J, Jiang N, Chen Q: A comparative study of small RNAs in Toxoplasma gondii of distinct genotypes. Parasit Vectors 2012, 5:186. BioMed Central Full Text
• [33]Wang PY, Yuan ZG, Petersen E, Li J, Zhang XX, Li XZ, Li HX, Lv ZC, Cheng T, Ren D, Yang GL, Lin RQ, Zhu XQ: Protective efficacy of a Toxoplasma gondii rhoptry protein 13 plasmid DNA vaccine in mice. Clin Vaccine Immunol 2012, 19(12):1916-1920.