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

Particle and Fibre Toxicology

Xing-Quan Zhu³  Qunli Zhao²  Gang Lu²  Hua Cong²  Huaiyu Zhou²  Yang Bai²  Guanghui Zhao²  Shenyi He¹  Min Meng¹ 

[1] State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, Peoples Republic of China;Department of Parasitology, Shandong University School of Medicine, Jinan, Shandong Province, 250012, Peoples Republic of China;College of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan Province, 650201, Peoples Republic of China

关键词: BALB/c mice;    Immunity;    DNA vaccine;    14-3-3;    SAG1;    Toxoplasma gondii;   

Others  :  1228827
DOI  :  10.1186/1756-3305-5-273

 received in 2012-10-28, accepted in 2012-11-14,  发布年份 2012

【 摘 要 】

Background

Toxoplasmosis, caused by an obligate intracellular protozoan parasite Toxoplasma gondii, has been a serious clinical and veterinary problem. Effective DNA vaccines against T. gondii can prevent and control the spread of toxoplasmosis, which is important for both human health and the farming industry. The T. gondii 14-3-3 protein has been proved to be antigenic and immunogenic and was a potential vaccine candidate against toxoplasmosis. In this study, we evaluated the immune responses induced by recombinant plasmids encoding T. gondii surface antigen 1 (SAG1) and 14-3-3 protein by immunizing BALB/c mice intramuscularly.

Methods

In the present study, BALB/c mice were randomly divided into five groups, including three experimental groups (pSAG1, p14-3-3 and pSAG1/14-3-3) and two control groups (PBS and pBudCE4.1), and were immunized intramuscularly three times. The levels of IgG antibodies and cytokine production in mouse sera were determined by enzyme-linked immunosorbent assays (ELISA). Two weeks after the last immunization, all mice were challenged intraperitoneally (i.p.) with 1×10⁴ tachyzoites of T. gondii and the survival time of mice was observed and recorded every day.

Results

Mice vaccinated with pSAG1, p14-3-3 or pSAG1/14-3-3 developed high levels of IgG2a and gamma interferon (IFN-γ) and low levels of interleukin-4 (IL-4) and interleukin-10 (IL-10) compared to control groups (PBS or pBudCE4.1), which suggested a modulated Th1 type immune response (P<0.05). After intraperitoneal challenge with 1×10⁴ tachyzoites of T. gondii (RH strain), the survival time of mice in experimental groups was longer than control groups (P<0.05). Mouse immunized with pSAG1/14-3-3 induced a higher level of IgG antibody response and significantly prolonged the survival time when compared with pSAG1 or p14-3-3 (P<0.05).

Conclusions

The study suggested that T. gondii 14-3-3 protein can induce effective immune responses in BALB/c mice and was a novel DNA vaccine candidate against toxoplasmosis, and the immune protective efficacy elicited by SAG1 gene was also demonstrated. Our results also showed multi-gene vaccine significantly enhanced immune responses and protective efficacy and was superior to the single-gene vaccine.

【 授权许可】

   
2012 Meng et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Dubey JP: The history of Toxoplasma gondii–the first 100 years. J Eukaryot Microbiol 2008, 55(6):467-475.
• [2]Sullivan WJ Jr, Jeffers V: Mechanisms of Toxoplasma gondii persistence and latency. FEMS Microbiol Rev 2012, 36(3):717-733.
• [3]Elmore SA, Jones JL, Conrad PA, Patton S, Lindsay DS, Dubey JP: Toxoplasma gondii: epidemiology, feline clinical aspects, and prevention. Trends Parasitol 2010, 26(4):190-196.
• [4]Dubey JP, Lindsay DS, Speer CA: Structures of Toxoplasma gondii tachyzoites, bradyzoites, and sporozoites and biology and development of tissue cysts. Clin Microbiol Rev 1998, 11(2):267-299.
• [5]Tenter AM, Heckeroth AR, Weiss LM: Toxoplasma gondii: from animals to humans. Int J Parasitol 2000, 30(12–13):1217-1258.
• [6]Munoz M, Liesenfeld O, Heimesaat MM: Immunology of Toxoplasma gondii. Immunol Rev 2011, 240(1):269-285.
• [7]Yang N, Mu MY, Li HK, Long M, He JB: Seroprevalence of Toxoplasma gondii infection in slaughtered chickens, ducks, and geese in Shenyang, northeastern China. Parasit Vectors 2012, 5(1):237. BioMed Central Full Text
• [8]Montoya JG, Liesenfeld O: Toxoplasmosis. Lancet 2004, 363(9425):1965-1976.
• [9]Weiss LM, Dubey JP: Toxoplasmosis: A history of clinical observations. Int J Parasitol 2009, 39(8):895-901.
• [10]Cenci-Goga BT, Rossitto PV, Sechi P, McCrindle CM, Cullor JS: Toxoplasma in animals, food, and humans: an old parasite of new concern. Foodborne Pathog Dis 2011, 8(7):751-762.
• [11]Rodriguez JB, Szajnman SH: New antibacterials for the treatment of toxoplasmosis; a patent review. Expert Opin Ther Pat 2012, 22(3):311-333.
• [12]Kur J, Holec-Gasior L, Hiszczynska-Sawicka E: Current status of toxoplasmosis vaccine development. Expert Rev Vaccines 2009, 8(6):791-808.
• [13]Kalinna BH: DNA vaccines for parasitic infections. Immunol Cell Biol 1997, 75(4):370-375.
• [14]Wilkins MF, O'Connell E, Te Punga WA: Toxoplasmosis in sheep III. Further evaluation of the ability of a live Toxoplasma gondii vaccine to prevent lamb losses and reduce congenital infection following experimental oral challenge. N Z Vet J 1988, 36(2):86-89.
• [15]Buxton D, Thomson K, Maley S, Wright S, Bos HJ: Vaccination of sheep with a live incomplete strain (S48) of Toxoplasma gondii and their immunity to challenge when pregnant. Vet Rec 1991, 129(5):89-93.
• [16]Buxton D, Innes EA: A commercial vaccine for ovine toxoplasmosis. Parasitology 1995, 110(Suppl):S11-S16.
• [17]Hiszczynska-Sawicka E, Holec-Gasior L, Kur J: DNA vaccines and recombinant antigens in prevention of Toxoplasma gondii infections–current status of the studies. Wiad Parazytol 2009, 55(2):125-139.
• [18]Jongert E, Roberts CW, Gargano N, Forster-Waldl E, Petersen E: Vaccines against Toxoplasma gondii: challenges and opportunities. Mem Inst Oswaldo Cruz 2009, 104(2):252-266.
• [19]Garcia JL: Vaccination concepts against Toxoplasma gondii. Expert Rev Vaccines 2009, 8(2):215-225.
• [20]Fu H, Subramanian RR, Masters SC: 14-3-3 proteins: structure, function, and regulation. Annu Rev Pharmacol Toxicol 2000, 40:617-647.
• [21]Aitken A, Collinge DB, van Heusden BP, Isobe T, Roseboom PH, Rosenfeld G, Soll J: 14-3-3 proteins: a highly conserved, widespread family of eukaryotic proteins. Trends Biochem Sci 1992, 17(12):498-501.
• [22]Koyama T, Ohsawa T, Shimada S, Omata Y, Xuan X, Inoue N, Maeda R, Mikami T, Saito A: A 14-3-3 protein homologue is expressed in feline enteroepithelial-stages of Toxoplasma gondii. Vet Parasitol 2001, 96(1):65-74.
• [23]Assossou O, Besson F, Rouault JP, Persat F, Brisson C, Duret L, Ferrandiz J, Mayencon M, Peyron F, Picot S: Subcellular localization of 14-3-3 proteins in Toxoplasma gondii tachyzoites and evidence for a lipid raft-associated form. FEMS Microbiol Lett 2003, 224(2):161-168.
• [24]Assossou O, Besson F, Rouault JP, Persat F, Ferrandiz J, Mayencon M, Peyron F, Picot S: Characterization of an excreted/secreted antigen form of 14-3-3 protein in Toxoplasma gondii tachyzoites. FEMS Microbiol Lett 2004, 234(1):19-25.
• [25]Schechtman D, Tarrab-Hazdai R, Arnon R: The 14-3-3 protein as a vaccine candidate against schistosomiasis. Parasite Immunol 2001, 23(4):213-217.
• [26]Menzies PI: Vaccination programs for reproductive disorders of small ruminants. Anim Reprod Sci 2012, 130(3–4):162-172.
• [27]Fachado A, Rodriguez A, Angel SO, Pinto DC, Vila I, Acosta A, Amendoeira RR, Lannes-Vieira J: Protective effect of a naked DNA vaccine cocktail against lethal toxoplasmosis in mice. Vaccine 2003, 21(13–14):1327-1335.
• [28]Beghetto E, Nielsen HV, Del Porto P, Buffolano W, Guglietta S, Felici F, Petersen E, Gargano N: A combination of antigenic regions of Toxoplasma gondii microneme proteins induces protective immunity against oral infection with parasite cysts. J Infect Dis 2005, 191(4):637-645.
• [29]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.
• [30]Siachoque H, Guzman F, Burgos J, Patarroyo ME, Gomez Marin JE: Toxoplasma gondii: immunogenicity and protection by P30 peptides in a murine model. Exp Parasitol 2006, 114(1):62-65.
• [31]Liu KY, Zhang DB, Wei QK, Li J, Li GP, Yu JZ: Biological role of surface Toxoplasma gondii antigen in development of vaccine. World J Gastroenterol 2006, 12(15):2363-2368.
• [32]Dautu G, Munyaka B, Carmen G, Zhang G, Omata Y, Xuenan X, Igarashi M: Toxoplasma gondii: DNA vaccination with genes encoding antigens MIC2, M2AP, AMA1 and BAG1 and evaluation of their immunogenic potential. Exp Parasitol 2007, 116(3):273-282.
• [33]Gong H, Zhou J, Liao M, Hatta T, Harnnoi T, Umemiya R, Inoue N, Xuan X, Fujisaki K: Characterization of a carboxypeptidase inhibitor from the tick Haemaphysalis longicornis. J Insect Physiol 2007, 53(10):1079-1087.
• [34]Zhou H, Min J, Zhao Q, Gu Q, Cong H, Li Y, He S: Protective immune response against Toxoplasma gondii elicited by a recombinant DNA vaccine with a novel genetic adjuvant. Vaccine 2012, 30(10):1800-1806.
• [35]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):5631-5636.
• [36]Cui X, Lei T, Yang D, Hao P, Li B, Liu Q: Toxoplasma gondii immune mapped protein-1 (TgIMP1) is a novel vaccine candidate against toxoplasmosis. Vaccine 2012, 30(13):2282-2287.
• [37]Pulendran B: Modulating TH1/TH2 responses with microbes, dendritic cells, and pathogen recognition receptors. Immunol Res 2004, 29(1–3):187-196.
• [38]Dupont CD, Christian DA, Hunter CA: Immune response and immunopathology during toxoplasmosis. Semin Immunopathol 2012. [Epub ahead of print]
• [39]Suzuki Y, Orellana MA, Schreiber RD, Remington JS: Interferon-gamma: the major mediator of resistance against Toxoplasma gondii. Science 1988, 240(4851):516-518.
• [40]Sher A, Collazzo C, Scanga C, Jankovic D, Yap G, Aliberti J: Induction and regulation of IL-12-dependent host resistance to Toxoplasma gondii. Immunol Res 2003, 27(2–3):521-528.
• [41]Liesenfeld O, Kosek J, Remington JS, Suzuki Y: Association of CD4+ T cell-dependent, interferon-gamma-mediated necrosis of the small intestine with genetic susceptibility of mice to peroral infection with Toxoplasma gondii. J Exp Med 1996, 184(2):597-607.
• [42]Liesenfeld O, Kang H, Park D, Nguyen TA, Parkhe CV, Watanabe H, Abo T, Sher A, Remington JS, Suzuki Y: TNF-alpha, nitric oxide and IFN-gamma are all critical for development of necrosis in the small intestine and early mortality in genetically susceptible mice infected perorally with Toxoplasma gondii. Parasite Immunol 1999, 21(7):365-376.
• [43]Liu Q, Shang L, Jin H, Wei F, Zhu XQ, Gao H: The protective effect of a Toxoplasma gondii SAG1 plasmid DNA vaccine in mice is enhanced with IL-18. Res Vet Sci 2010, 89(1):93-97.
• [44]Angus CW, Klivington-Evans D, Dubey JP, Kovacs JA: Immunization with a DNA plasmid encoding the SAG1 (P30) protein of Toxoplasma gondii is immunogenic and protective in rodents. J Infect Dis 2000, 181(1):317-324.
• [45]Sun XM, Zou J, AE A, 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
• [46]Henriquez FL, Woods S, Cong H, McLeod R, Roberts CW: Immunogenetics of Toxoplasma gondii informs vaccine design. Trends Parasitol 2010, 26(11):550-555.
• [47]Dzierszinski FS, Hunter CA: Advances in the use of genetically engineered parasites to study immunity to Toxoplasma gondii. Parasite Immunol 2008, 30(4):235-244.
• [48]Schneider J, Gilbert SC, Blanchard TJ, Hanke T, Robson KJ, Hannan CM, Becker M, Sinden R, Smith GL, Hill AV: Enhanced immunogenicity for CD8+ T cell induction and complete protective efficacy of malaria DNA vaccination by boosting with modified vaccinia virus Ankara. Nat Med 1998, 4(4):397-402.