• 1.

    Dubey JP, 2010. Toxoplasmosis of Animals and Humans. Boca Raton, FL: CRC Press, 313.

  • 2.

    Jackson MH, Hutchison WM, 1989. The prevalence and source of Toxoplasma infection in the environment. Adv Parasitol 28: 55105.

  • 3.

    Bhopale GM, 2003. Development of a vaccine for toxoplasmosis: current status. Microbes Infect 5: 457462.

  • 4.

    Lyons RE, McLeod R, Roberts CW, 2002. Toxoplasma gondii tachyzoite-bradyzoite interconversion. Trends Parasitol 18: 198201.

  • 5.

    Black MW, Boothroyd JC, 2000. Lytic cycle of Toxoplasma gondii. Microbiol Mol Biol Rev 64: 607623.

  • 6.

    Kur J, Holec-Gasior L, Hiszczynska-Sawicka E, 2009. Current status of toxoplasmosis vaccine development. Expert Rev Vaccines 8: 791808.

  • 7.

    Corr M, Tighe H, 1997. Plasmid DNA vaccination: mechanism of antigen presentation. Springer Semin Immunopathol 19: 139145.

  • 8.

    Bessieres MH, Swierczynski B, Cassaing S, Miedouge M, Olle P, Seguela JP, Pipy B, 1997. Role of IFN-gamma, TNF-alpha, IL4 and IL10 in the regulation of experimental Toxoplasma gondii infection. J Eukaryot Microbiol 44: 87S.

    • Search Google Scholar
    • Export Citation
  • 9.

    Gazzinelli RT, Hakim FT, Hieny S, Shearer GM, Sher A, 1991. Synergistic role of CD4+ and CD8+ T lymphocytes in IFN-gamma production and protective immunity induced by an attenuated Toxoplasma gondii vaccine. J Immunol 146: 286292.

    • Search Google Scholar
    • Export Citation
  • 10.

    Nam HW, 2009. GRA proteins of Toxoplasma gondii: maintenance of host-parasite interactions across the parasitophorous vacuolar membrane. Korean J Parasitol 47 (Suppl): S29S37.

    • Search Google Scholar
    • Export Citation
  • 11.

    Capron A, Dessaint JP, 1988. Vaccination against parasitic diseases: some alternative concepts for the definition of protective antigens. Ann Inst Pasteur Immunol 139: 109117.

    • Search Google Scholar
    • Export Citation
  • 12.

    Cesbron-Delauw MF, Capron A, 1993. Excreted/secreted antigens of Toxoplasma gondii–their origin and role in the host-parasite interaction. Res Immunol 144: 4144.

    • Search Google Scholar
    • Export Citation
  • 13.

    Hiszczynska-Sawicka E, Oledzka G, Holec-Gasior L, Li H, Xu JB, Sedcole R, Kur J, Bickerstaffe R, Stankiewicz M, 2011. Evaluation of immune responses in sheep induced by DNA immunization with genes encoding GRA1, GRA4, GRA6 and GRA7 antigens of Toxoplasma gondii. Vet Parasitol 177: 281289.

    • Search Google Scholar
    • Export Citation
  • 14.

    Scorza T, D'Souza S, Laloup M, Dewit J, De Braekeleer J, Verschueren H, Vercammen M, Huygen K, Jongert E, 2003. A GRA1 DNA vaccine primes cytolytic CD8(+) T cells to control acute Toxoplasma gondii infection. Infect Immun 71: 309316.

    • Search Google Scholar
    • Export Citation
  • 15.

    Sun XM, Zou J, Saeed EAA, Yan WC, Liu XY, Suo X, Wang H, Chen QJ, 2011. DNA vaccination with a gene encoding Toxoplasma gondii GRA6 induces partial protection against toxoplasmosis in BALB/c mice. Parasit Vectors 4: 213.

    • Search Google Scholar
    • Export Citation
  • 16.

    Feng P, Park J, Lee BS, Lee SH, Bram RJ, Jung JU, 2002. Kaposi's sarcoma-associated herpesvirus mitochondrial K7 protein targets a cellular calcium-modulating cyclophilin ligand to modulate intracellular calcium concentration and inhibit apoptosis. J Virol 76: 1149111504.

    • Search Google Scholar
    • Export Citation
  • 17.

    Tilley M, Fichera ME, Jerome ME, Roos DS, White MW, 1997. Toxoplasma gondii sporozoites form a transient parasitophorous vacuole that is impermeable and contains only a subset of dense-granule proteins. Infect Immun 65: 45984605.

    • Search Google Scholar
    • Export Citation
  • 18.

    Zhou H, Gu Q, Zhao Q, Zhang J, Cong H, Li Y, He S, 2007. Toxoplasma gondii: expression and characterization of a recombinant protein containing SAG1 and GRA2 in Pichia pastoris. Parasitol Res 100: 829835.

    • Search Google Scholar
    • Export Citation
  • 19.

    Cao A, Liu Y, Wang J, Li X, Wang S, Zhao Q, Cong H, He S, Zhou H, 2015. 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 33: 67576762.

    • Search Google Scholar
    • Export Citation
  • 20.

    Liu S, Shi L, Cheng YB, Fan GX, Ren HX, Yuan YK, 2009. Evaluation of protective effect of multi-epitope DNA vaccine encoding six antigen segments of Toxoplasma gondii in mice. Parasitol Res 105: 267274.

    • Search Google Scholar
    • Export Citation
  • 21.

    Naserifar R, Ghaffarifar F, Dalimi A, Sharifi Z, Solhjoo K, Hosseinian Khosroshahi K, 2015. Evaluation of immunogenicity of cocktail DNA vaccine containing plasmids encoding complete GRA5, SAG1, and ROP2 antigens of Toxoplasma gondii in BALB/C mice. Iran J Parasitol 10: 590598.

    • Search Google Scholar
    • Export Citation
  • 22.

    Xue M, He S, Cui Y, Yao Y, Wang H, 2008. Evaluation of the immune response elicited by multi-antigenic DNA vaccine expressing SAG1, ROP2 and GRA2 against Toxoplasma gondii. Parasitol Int 57: 424429.

    • Search Google Scholar
    • Export Citation
  • 23.

    Ching XT, Fong MY, Lau YL, 2016. Evaluation of immunoprotection conferred by the subunit vaccines of GRA2 and GRA5 against acute toxoplasmosis in BALB/c mice. Front Microbiol 7: 609.

    • Search Google Scholar
    • Export Citation
  • 24.

    Ching X-T, Lau Y-L, Fong M-Y, 2015. Heterologous expression of Toxoplasma gondii dense granule protein 2 and 5. Southeast Asian J Trop Med Public Health 46: 375387.

    • Search Google Scholar
    • Export Citation
  • 25.

    Kaplan EL, Meier P, 1958. Nonparametric estimation from incomplete observations. J Am Stat Assoc 53: 457481.

  • 26.

    Kim K, Bulow R, Kampmeier J, Boothroyd JC, 1994. Conformationally appropriate expression of the Toxoplasma antigen SAG1 (p30) in CHO cells. Infect Immun 62: 203209.

    • Search Google Scholar
    • Export Citation
  • 27.

    Abdizadeh R, Maraghi S, Ghadiri AA, Tavalla M, Shojaee S, 2015. Cloning and expression of major surface antigen 1 gene of Toxoplasma gondii RH strain using the expression vector pVAX1 in Chinese hamster ovary cells. Jundishapur J Microbiol 8: e22570.

    • Search Google Scholar
    • Export Citation
  • 28.

    Sonaimuthu P, Ching XT, Fong MY, Kalyanasundaram R, Lau YL, 2016. Induction of protective immunity against toxoplasmosis in BALB/c mice vaccinated with Toxoplasma gondii Rhoptry-1. Front Microbiol 7: 808.

    • Search Google Scholar
    • Export Citation
  • 29.

    Parthasarathy S, Fong MY, Ramaswamy K, Lau YL, 2013. Protective immune response in BALB/c mice induced by DNA vaccine of the ROP8 gene of Toxoplasma gondii. Am J Trop Med Hyg 88: 883887.

    • Search Google Scholar
    • Export Citation
  • 30.

    Ihara F, Nishikawa Y, 2014. Starvation of low-density lipoprotein-derived cholesterol induces bradyzoite conversion in Toxoplasma gondii. Parasit Vectors 7: 248.

    • Search Google Scholar
    • Export Citation
  • 31.

    Kunert R, Vorauer-Uhl K, 2012. Strategies for efficient transfection of CHO-cells with plasmid DNA. Methods Mol Biol 801: 213226.

  • 32.

    Hartley JL, 2012. Why proteins in mammalian cells? Methods Mol Biol 801: 112.

  • 33.

    Babaie J, Sadeghiani G, Golkar M, 2011. Construction and in vitro expression analyses of a DNA plasmid encoding dense granule GRA5 antigen of Toxoplasma gondii. Avicenna J Med Biotechnol 3: 135141.

    • Search Google Scholar
    • Export Citation
  • 34.

    Golkar M, Shokrgozar MA, Rafati S, Sadaie MR, Assmar M, 2005. Construction, expression and preliminary immunological evaluation of a DNA plasmid encoding the GRA2 protein of Toxoplasma gondii. Iran Biomed J 9: 18.

    • Search Google Scholar
    • Export Citation
  • 35.

    Zhou H, Min J, Zhao Q, Gu Q, Cong H, Li Y, He S, 2012. Protective immune response against Toxoplasma gondii elicited by a recombinant DNA vaccine with a novel genetic adjuvant. Vaccine 30: 18001806.

    • Search Google Scholar
    • Export Citation
  • 36.

    Chen J, Huang SY, Li ZY, Yuan ZG, Zhou DH, Petersen E, Zhang NZ, Zhu XQ, 2013. Protective immunity induced by a DNA vaccine expressing eIF4A of Toxoplasma gondii against acute toxoplasmosis in mice. Vaccine 31: 17341739.

    • Search Google Scholar
    • Export Citation
  • 37.

    Hiszczynska-Sawicka E, Li H, Xu JB, Oledzka G, Kur J, Bickerstaffe R, Stankiewicz M, 2010. Comparison of immune response in sheep immunized with DNA vaccine encoding Toxoplasma gondii GRA7 antigen in different adjuvant formulations. Exp Parasitol 124: 365372.

    • Search Google Scholar
    • Export Citation
  • 38.

    Ismael AB, Sekkai D, Collin C, Bout D, Mevelec MN, 2003. The MIC3 gene of Toxoplasma gondii is a novel potent vaccine candidate against toxoplasmosis. Infect Immun 71: 62226228.

    • Search Google Scholar
    • Export Citation
  • 39.

    Qu D, Han J, Du A, 2013. Evaluation of protective effect of multiantigenic DNA vaccine encoding MIC3 and ROP18 antigen segments of Toxoplasma gondii in mice. Parasitol Res 112: 25932599.

    • Search Google Scholar
    • Export Citation
  • 40.

    Tao Q, Fang R, Zhang W, Wang Y, Cheng J, Li Y, Fang K, Khan MK, Hu M, Zhou Y, Zhao J, 2013. Protective immunity induced by a DNA vaccine-encoding Toxoplasma gondii microneme protein 11 against acute toxoplasmosis in BALB/c mice. Parasitol Res 112: 28712877.

    • Search Google Scholar
    • Export Citation
  • 41.

    Wu XN, Lin J, Lin X, Chen J, Chen ZL, Lin JY, 2012. Multicomponent DNA vaccine-encoding Toxoplasma gondii GRA1 and SAG1 primes: anti-Toxoplasma immune response in mice. Parasitol Res 111: 20012009.

    • Search Google Scholar
    • Export Citation
  • 42.

    Yuan ZG, Zhang XX, Lin RQ, Petersen E, He S, Yu M, He XH, Zhou DH, He Y, Li HX, Liao M, Zhu XQ, 2011. Protective effect against toxoplasmosis in mice induced by DNA immunization with gene encoding Toxoplasma gondii ROP18. Vaccine 29: 66146619.

    • Search Google Scholar
    • Export Citation
  • 43.

    Yuan ZG, Zhang XX, He XH, Petersen E, Zhou DH, He Y, Lin RQ, Li XZ, Chen XL, Shi XR, Zhong XL, Zhang B, Zhu XQ, 2011. Protective immunity induced by Toxoplasma gondii rhoptry protein 16 against toxoplasmosis in mice. Clin Vaccine Immunol 18: 119124.

    • Search Google Scholar
    • Export Citation
  • 44.

    Rosenberg C, De Craeye S, Jongert E, Gargano N, Beghetto E, Del Porto P, Vorup-Jensen T, Petersen E, 2009. Induction of partial protection against infection with Toxoplasma gondii genotype II by DNA vaccination with recombinant chimeric tachyzoite antigens. Vaccine 27: 24892498.

    • Search Google Scholar
    • Export Citation
  • 45.

    Chu RS, Targoni OS, Krieg AM, Lehmann PV, Harding CV, 1997. CpG oligodeoxynucleotides act as adjuvants that switch on T helper 1 (Th1) immunity. J Exp Med 186: 16231631.

    • Search Google Scholar
    • Export Citation
  • 46.

    Klinman DM, Yamshchikov G, Ishigatsubo Y, 1997. Contribution of CpG motifs to the immunogenicity of DNA vaccines. J Immunol 158: 36353639.

  • 47.

    Roman M, Martin-Orozco E, Goodman JS, Nguyen MD, Sato Y, Ronaghy A, Kornbluth RS, Richman DD, Carson DA, Raz E, 1997. Immunostimulatory DNA sequences function as T helper-1-promoting adjuvants. Nat Med 3: 849854.

    • Search Google Scholar
    • Export Citation
  • 48.

    Sato Y, Roman M, Tighe H, Lee D, Corr M, Nguyen MD, Silverman GJ, Lotz M, Carson DA, Raz E, 1996. Immunostimulatory DNA sequences necessary for effective intradermal gene immunization. Science 273: 352354.

    • Search Google Scholar
    • Export Citation
  • 49.

    York IA, Goldberg AL, Mo XY, Rock KL, 1999. Proteolysis and class I major histocompatibility complex antigen presentation. Immunol Rev 172: 4966.

    • Search Google Scholar
    • Export Citation
  • 50.

    Beghetto E, Nielsen HV, Del Porto P, Buffolano W, Guglietta S, Felici F, Petersen E, Gargano N, 2005. A combination of antigenic regions of Toxoplasma gondii microneme proteins induces protective immunity against oral infection with parasite cysts. J Infect Dis 191: 637645.

    • Search Google Scholar
    • Export Citation
  • 51.

    Dautu G, Munyaka B, Carmen G, Zhang G, Omata Y, Xuenan X, Igarashi M, 2007. Toxoplasma gondii: DNA vaccination with genes encoding antigens MIC2, M2AP, AMA1 and BAG1 and evaluation of their immunogenic potential. Exp Parasitol 116: 273282.

    • Search Google Scholar
    • Export Citation
  • 52.

    Ghaffarifar F, Naserifar R, Jafari Madrak M, 2014. Eukaryotic plasmids with Toxoplasma gondii dense granule antigen (GRA 5) and Microneme 3 (MIC3) genes as a cocktail DNA vaccine and evaluation of immune responses in BALB/C Mice. J Clin Med Genom 3: 2.

    • Search Google Scholar
    • Export Citation
  • 53.

    Sayles PC, Gibson GW, Johnson LL, 2000. B cells are essential for vaccination-induced resistance to virulent Toxoplasma gondii. Infect Immun 68: 10261033.

    • Search Google Scholar
    • Export Citation
  • 54.

    Johnson LL, Sayles PC, 2002. Deficient humoral responses underlie susceptibility to Toxoplasma gondii in CD4-deficient mice. Infect Immun 70: 185191.

    • Search Google Scholar
    • Export Citation
  • 55.

    Frenkel JK, Taylor DW, 1982. Toxoplasmosis in immunoglobulin M-suppressed mice. Infect Immun 38: 360367.

  • 56.

    Johnson AM, McDonald PJ, Neoh SH, 1983. Monoclonal antibodies to Toxoplasma cell membrane surface antigens protect mice from toxoplasmosis. J Protozool 30: 351356.

    • Search Google Scholar
    • Export Citation
  • 57.

    McLeod R, Eisenhauer P, Mack D, Brown C, Filice G, Spitalny G, 1989. Immune responses associated with early survival after peroral infection with Toxoplasma gondii. J Immunol 142: 32473255.

    • Search Google Scholar
    • Export Citation
  • 58.

    Roberts CW, Ferguson DJ, Jebbari H, Satoskar A, Bluethmann H, Alexander J, 1996. Different roles for interleukin-4 during the course of Toxoplasma gondii infection. Infect Immun 64: 897904.

    • Search Google Scholar
    • Export Citation
  • 59.

    Gazzinelli RT, Wysocka M, Hieny S, Scharton-Kersten T, Cheever A, Kuhn R, Muller W, Trinchieri G, Sher A, 1996. In the absence of endogenous IL-10, mice acutely infected with Toxoplasma gondii succumb to a lethal immune response dependent on CD4+ T cells and accompanied by overproduction of IL-12, IFN-gamma and TNF-alpha. J Immunol 157: 798805.

    • Search Google Scholar
    • Export Citation
 
 
 

 

 
 
 

 

 

 

 

 

 

Evaluation of the Protective Effect of Deoxyribonucleic Acid Vaccines Encoding Granule Antigen 2 and 5 Against Acute Toxoplasmosis in BALB/c Mice

View More View Less
  • 1 Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

Toxoplasma gondii infects a broad range of warm-blooded hosts, including humans. Important clinical manifestations include encephalitis in immunocompromised patients as well as miscarriage and fetal damage during early pregnancy. Toxoplasma gondii dense granule antigen 2 and 5 (GRA2 and GRA5) are essential for parasitophorous vacuole development of the parasite. To evaluate the potential of GRA2 and GRA5 as recombinant DNA vaccine candidates, these antigens were cloned into eukaryotic expression vector (pcDNA 3.1C) and evaluated in vaccination experiments. Recombinant DNA vaccines constructed with genes encoding GRAs were validated in Chinese hamster ovary cells before evaluation using lethal challenge of the virulent T. gondii RH strain in BALB/c mice. The DNA vaccines of pcGRA2 and pcGRA5 elicited cellular-mediated immune response with significantly higher levels of interferon-gamma, interleukin-2 (IL-2), IL-4, and IL-10 (P < 0.05) compared with controls. A mixed T-helper cell 1 (Th1)/Th2 response was associated with slightly prolonged survival. These findings provide evidence that DNA vaccination with GRA2 and GRA5 is associated with Th1-like cell-mediated immune responses. It will be worthwhile to construct recombinant multiantigen combining full-length GRA2 or/and GRA5 with various antigenic proteins such as the surface antigens and rhoptry antigens to improve vaccination efficacy.

Author Notes

* Address correspondence to Yee Ling Lau, Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia. E-mail: lauyeeling@um.edu.my

Financial support: This research project was supported by University of Malaya High Impact Research (HIR) Grant UM-MOHE (UM.C/HIR/MOHE/MED/16) from the Ministry of Higher Education, Malaysia.

Authors' addresses: Xiao Teng Ching, Mun Yik Fong, and Yee Ling Lau, Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia, E-mails: cxteng85@yahoo.com, fongmy@um.edu.my, and lauyeeling@um.edu.my.

Save