Abstract
Toxoplasmosis is one of the most common parasitic infections worldwide. An effective vaccine against human and animal toxoplasmosis is still needed to control this parasitosis. The polymorphic rhoptry proteins, ROP5 and ROP18, secreted by Toxoplasma gondii during the invasion of the host cell have been recently considered as promising vaccine antigens, as they appear to be the major determinants of T. gondii virulence in mice. The goal of this study was to evaluate their immunogenic and immunoprotective activity after their administration (separately or both recombinant proteins together) with the poly I:C as an adjuvant. Immunization of BALB/c and C3H/HeOuJ mice generated both cellular and humoral specific immune responses with some predominance of IgG1 antibodies. The spleen cells derived from vaccinated animals reacted to the parasite’s native antigens. Furthermore, the immunization led to a partial protection against acute and chronic toxoplasmosis. These findings confirm the previous assumptions about ROP5 and ROP18 antigens as valuable components of a subunit vaccine against toxoplasmosis.
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Abdollahi SH, Ayoobi F, Khorramdelazad H, Hassanshahi G, Ahmadabadi BN, Rezayati M, Ravary A, Shamsizadeh A, Arababadi MK (2013) Interleukin-10 serum levels after vaccination with in vivo prepared Toxoplasma gondii excreted/secreted antigens. Oman Med J 28:112–5
Araujo FG, Williams DM, Grumet FC, Remington JS (1976) Strain-dependent differences in murine susceptibility to toxoplasma. Infect Immun 13:1528–1530
Behnke MS, Khan A, Wootton JC, Dubey JP, Tang K, Sibley LD (2011) Virulence differences in Toxoplasma mediated by amplification of a family of polymorphic pseudokinases. Proc Natl Acad Sci U S A 108:9631–9636
Blader IJ, Saeij JP (2009) Communication between Toxoplasma gondii and its host: impact on parasite growth, development, immune evasion, and virulence. APMIS 117:458–476
Borges O, Borchard G, de Sousa A, Junginger HE, Cordeiro-da-Silva A (2007) Induction of lymphocytes activated marker CD69 following exposure to chitosan and alginate biopolymers. Int J Pharm 337:254–264
Chen J, Li ZY, Petersen E, Huang SY, Zhou DH, Zhu XQ (2015) DNA vaccination with genes encoding Toxoplasma gondii antigens ROP5 and GRA15 induces protective immunity against toxoplasmosis in Kunming mice. Expert Rev Vaccines 14:617–624
Denkers EY, Gazzinelli RT (1998) Regulation and function of T-cell-mediated immunity during Toxoplasma gondii infection. Clin Microbiol Rev 11:569–588
Dlugonska H (2008) Toxoplasma rhoptries: unique secretory organelles and source of promising vaccine proteins for immunoprevention of toxoplasmosis. J Biomed Biotechnol 2008:632424
Dubey JP (2009) History of the discovery of the life cycle of Toxoplasma gondii. Int J Parasitol 39:877–882
Dupont CD, Christian DA, Hunter CA (2012) Immune response and immunopathology during toxoplasmosis. Semin Immunopathol 34:793–813
Dziadek B, Gatkowska J, Brzostek A, Dziadek J, Dzitko K, Długonska H (2009) Toxoplasma gondii: the immunogenic and protective efficacy of recombinant ROP2 and ROP4 rhoptry proteins in murine experimental toxoplasmosis. Exp Parasitol 123:81–89
Dziadek B, Gatkowska J, Brzostek A, Dziadek J, Dzitko K, Grzybowski M, Dlugonska H (2011) Evaluation of three recombinant multi-antigenic vaccines composed of surface and secretory antigens of Toxoplasma gondii in murine models of experimental toxoplasmosis. Vaccine 29:821–830
Dziadek B, Gatkowska J, Grzybowski M, Dziadek J, Dzitko K, Dlugonska H (2012) 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 131:133–138
El Hajj H, Demey E, Poncet J, Lebrun M, Wu B, Galéotti N, Fourmaux MN, Mercereau-Puijalon O, Vial H, Labesse G, Dubremetz JF (2006) The ROP2 family of Toxoplasma gondii rhoptry proteins: proteomic and genomic characterization and molecular modeling. Proteomics 6:5773–5784
El Hajj H, Lebrun M, Arold ST, Vial H, Labesse G, Dubremetz JF (2007) ROP18 is a rhoptry kinase controlling the intracellular proliferation of Toxoplasma gondii. PLoS Pathog 3, e14
Etheridge RD, Alaganan A, Tang K, Lou HJ, Turk BE, Sibley LD (2014) The Toxoplasma pseudokinase ROP5 forms complexes with ROP18 and ROP17 kinases that synergize to control acute virulence in mice. Cell Host Microbe 15:537–50
Fentress SJ, Sibley LD (2011) The secreted kinase ROP18 defends Toxoplasma’s border. Bioessays 33:693–700
Fentress SJ, Behnke MS, Dunay IR, Mashayekhi M, Rommereim LM, Fox BA, Bzik DJ, Taylor GA, Turk BE, Lichti CF, Townsend RR, Qiu W, Hui R, Beatty WL, Sibley LD (2010) Phosphorylation of immunity-related GTPases by a Toxoplasma gondii-secreted kinase promotes macrophage survival and virulence. Cell Host Microbe 8:484–495
Filisetti D, Candolfi E (2004) Immune response to Toxoplasma gondii. Ann Ist Super Sanita 40:71–80
Gatkowska J, Hiszczynska-Sawicka E, Kur J, Holec L, Długonska H (2006) Toxoplasma gondii: an evaluation of diagnostic value of recombinant antigens in a murine model. Exp Parasitol 114:220–227
Grzybowski MM, Dziadek B, Dziadek J, Gatkowska J, Dzitko K, Długońska H (2015) Toxoplasma gondii: cloning. expression and immunoreactivity of recombinant ROP5 and ROP18 antigens. Exp Parasitol 150:1–6
Hiszczyńska-Sawicka E, Gatkowska JM, Grzybowski MM, Długońska H (2014) Veterinary vaccines against toxoplasmosis. Parasitology 141:1365–1378
Innes EA, Bartley PM, Maley S, Katzer F, Buxton D (2009) Veterinary vaccines against Toxoplasma gondii. Mem Inst Oswaldo Cruz 104:246–51
Jensen KD, Camejo A, Melo MB, Cordeiro C, Julien L, Grotenbreg GM, Frickel EM, Ploegh HL, Young L, Saeij JP (2015) Toxoplasma gondii superinfection and virulence during secondary infection correlate with the exact ROP5/ROP18 allelic combination. MBio 6, e02280
Jongert E, Roberts CW, Gargano N, Förster-Waldl E, Petersen E (2009) Vaccines against Toxoplasma gondii: challenges and opportunities. Mem Inst Oswaldo Cruz 104:252–66
Khan A, Taylor S, Ajioka JW, Rosenthal BM, Sibley LD (2009) Selection at a single locus leads to widespread expansion of Toxoplasma gondii lineages that are virulent in mice. PLoS Genet 5, e1000404
Li XZ, Wang XH, Xia LJ, Weng YB, Hernandez JA, Tu LQ, Li LT, Li SJ, Yuan ZG (2015) Protective efficacy of recombinant canine adenovirus type-2 expressing TgROP18 (CAV-2-ROP18) against acute and chronic Toxoplasma gondii infection in mice. BMC Infect Dis 15:114
Montoya JG, Liesenfeld O (2004) Toxoplasmosis. Lancet 363:1965–1976
Mosmann TR, Coffman RL (1989) TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional properties. Annu Rev Immunol 7:145–173
Niedelman W, Gold DA, Rosowski EE, Sprokholt JK, Lim D, Farid Arenas A, Melo MB, Spooner E, Yaffe MB, Saeij JP (2012) The rhoptry proteins ROP18 and ROP5 mediate Toxoplasma gondii evasion of the murine, but not the human, interferon-gamma response. PLoS Pathog 8, e1002784
Pifer R, Yarovinsky F (2011) Innate responses to Toxoplasma gondii in mice and humans. Trends Parasitol 27:388–93
Qiu W, Wernimont A, Tang K, Taylor S, Lunin V, Schapira M, Fentress S, Hui R, Sibley LD (2009) Novel structural and regulatory features of rhoptry secretory kinases in Toxoplasma gondii. EMBO J 28:969–79
Qu D, Han J, Du A (2013a) Enhancement of protective immune response to recombinant Toxoplasma gondii ROP18 antigen by ginsenoside Re. Exp Parasitol 135:234–239
Qu D, Han J, Du A (2013b) Evaluation of protective effect of multiantigenic DNA vaccine encoding MIC3 and ROP18 antigen segments of Toxoplasma gondii in mice. Parasitol Res 112:2593–2599
Reese ML, Boyle JP (2012) Virulence without catalysis: how can a pseudokinase affect host cell signaling? Trends Parasitol 28:53–57
Reese ML, Zeiner GM, Saeij JP, Boothroyd JC, Boyle JP (2011) Polymorphic family of injected pseudokinases is paramount in Toxoplasma virulence. Proc Natl Acad Sci U S A 108:9625–9630
Reese ML, Shah N, Boothroyd JC (2014) The Toxoplasma pseudokinase ROP5 is an allosteric inhibitor of the immunity-related GTPases. J Biol Chem 289:27849–27858
Robert-Gangneux F, Dardé ML (2012) Epidemiology of and diagnostic strategies for toxoplasmosis. Clin Microbiol Rev 25:264–296
Roberts CW, McLeody R, Henriquez FL, Alexander J (2014) Vaccination against toxoplasmosis: current status and future prospects. In: Weiss LM, Kim K (ed) Toxoplasma gondii. The model apicomplexan – perspectives and methods, 2nd edn. Elsevier, pp 995–1045
Saeij JP, Boyle JP, Coller S, Taylor S, Sibley LD, Brooke-Powell ET, Ajioka JW, Boothroyd JC (2006) Polymorphic secreted kinases are key virulence factors in toxoplasmosis. Science 314:1780–1783
Salem ML, Kadima AN, Cole DJ, Gillanders WE (2005) Defining the antigen-specific T-cell response to vaccination and poly (I: C)/TLR3 signaling: evidence of enhanced primary and memory CD8 T-cell responses and antitumor immunity. J Immunother 28:220–228
Sayles PC, Gibson GW, Johnson LL (2000) B cells are essential for vaccination-induced resistance to virulent Toxoplasma gondii. Infect Immun 68:1026–1033
Selleck EM, Fentress SJ, Beatty WL, Degrandi D, Pfeffer K, Virgin HW 4th, Macmicking JD, Sibley LD (2013) Guanylate-binding protein 1 (Gbp1) contributes to cell-autonomous immunity against Toxoplasma gondii. PLoS Pathog 9, e1003320
Sheffield P, Garrard S, Derewenda Z (1999) Overcoming expression and purification problems of RhoGDI using a family of “parallel” expression vectors. Protein Expr Purif 15:34–39
Steinfeldt T, Könen-Waisman S, Tong L, Pawlowski N, Lamkemeyer T, Sibley LD, Hunn JP, Howard JC (2010) Phosphorylation of mouse immunity-related GTPase (IRG) resistance proteins is an evasion strategy for virulent Toxoplasma gondii. PLoS Biol 8, e1000576
Sturge CR, Yarovinsky F (2014) Complex immune cell interplay in the gamma interferon response during Toxoplasma gondii infection. Infect Immun 82:3090–3097
Sullivan WJ Jr, Smith AT, Joyce BR (2009) Understanding mechanisms and the role of differentiation in pathogenesis of Toxoplasma gondii: a review. Mem Inst Oswaldo Cruz 104:155–61
Suzuki Y, Orellana MA, Schreiber RD, Remington JS (1988) Interferon-gamma: the major mediator of resistance against Toxoplasma gondii. Science 240:516–518
Taylor S, Barragan A, Su C, Fux B, Fentress SJ, Tang K, Beatty WL, Hajj HE, Jerome M, Behnke MS, White M, Wootton JC, Sibley LD (2006) A secreted serine-threonine kinase determines virulence in the eukaryotic pathogen Toxoplasma gondii. Science 314:1776–1780
Tenter AM, Heckeroth AR, Weiss LM (2000) Toxoplasma gondii: from animals to humans. Int J Parasitol 30:1217–58
Vercammen M, Scorza T, Huygen K, De Braekeleer J, Diet R, Jacobs D, Saman E, Verschueren H (2000) DNA vaccination with genes encoding Toxoplasma gondii antigens GRA1, GRA7, and ROP2 induces partially protective immunity against lethal challenge in mice. Infect Immun 68:38–45
Verma R, Khanna P (2013) Development of Toxoplasma gondii vaccine: a global challenge. Hum Vaccin Immunother 9:291–293
Wilson EH, Wille-Reece U, Dzierszinski F, Hunter CA (2005) A critical role for IL-10 in limiting inflammation during toxoplasmic encephalitis. J Neuroimmunol 165:63–74
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 29:6614–6619
Zhang NZ, Chen J, Wang M, Petersen E, Zhu XQ (2013) Vaccines against Toxoplasma gondii: new developments and perspectives. Expert Rev Vaccines 12:1287–1299
Zheng B, Lu S, Tong Q, Kong Q, Lou D (2013) The virulence-related rhoptry protein 5 (ROP5) of Toxoplasma gondii is a novel vaccine candidate against toxoplasmosis in mice. Vaccine 31:4578–84
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This project was funded by the National Science Centre (Poland) on the basis of the decision DEC-2011/03/N/NZ6/04655.
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Grzybowski, M.M., Dziadek, B., Gatkowska, J.M. et al. Towards vaccine against toxoplasmosis: evaluation of the immunogenic and protective activity of recombinant ROP5 and ROP18 Toxoplasma gondii proteins. Parasitol Res 114, 4553–4563 (2015). https://doi.org/10.1007/s00436-015-4701-y
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DOI: https://doi.org/10.1007/s00436-015-4701-y