Abstract
Gametocyte proteins are being explored as potential vaccine candidates against Eimeria sp. in chicken since they are the components of the resilient oocyst wall. The aim of this study was to investigate the immunoprophylactic efficacy of recombinant Eimeria tenella gametocyte antigen 22 (EtGam22) in chickens against homologous oocyst challenge. Broiler chicks were subcutaneously immunized individually with 100 μg of recombinant EtGam22 adjuvanted with Montanide ISA 71 VG at 7 days of age and boosted 2 weeks later. The immunized chickens were challenged individually with 1 × 104 sporulated oocysts of E. tenella 1 week post-booster immunization. The anti-EtGam22 IgY and serum cytokine response was measured post-immunization. The results showed that the anti-EtGam22 IgY antibody, serum IFN-γ, IL-2, TGF-β, and IL-4 levels in chickens vaccinated with recombinant protein were significantly increased post-immunization as compared to unimmunized challenged controls (P < 0.05). The peripheral blood lymphocyte proliferation activity was also found significantly higher in EtGam22-immunized group on day 28, i.e., pre-challenge (P < 0.05). Upon homologous oocyst challenge, chickens immunized with rEtGam22 exhibited a significant drop in the total oocyst output per bird (246.78 ± 36.9 × 106, 45.23% reduction) and a significantly higher weight gain (497.7 ± 19.2 g) as compared to unimmunized challenged controls. Taken together, these data indicate that EtGam22 is a potent immunogen for use as a subunit vaccine against cecal coccidiosis in chickens as it induces a diverse and robust immune response involving multiple cytokines and strong antibody titers.
Similar content being viewed by others
References
Belli SI, Witcombe D, Wallach MG, Smith NC (2002) Functional genomics of gam56: characterisation of the role of a 56 kilodalton sexual stage antigen in oocyst wall formation in Eimeria maxima. J Parasitol 32:1727–1737
Belli SI, Wallach MG, Smith NC (2003) Cloning and characterization of the 82 kDa tyrosine-rich sexual stage glycoprotein, GAM82, and its role in oocyst wall formation in the apicomplexan parasite, Eimeria maxima. Gene 307:201–212
Belli SI, Mai K, Skene CD, Gleeson MT, Witcombe DM, Katrib M, Finger A, Wallach MG, Smith NC (2004) Characterisation of the antigenic and immunogenic properties of bacterially expressed, sexual stage antigens of the coccidian parasite, Eimeria maxima. Vaccine 22:4316–4325
Belli SI, Smith NC, Ferguson DJ (2006) The coccidian oocyst: a tough nut to crack! Trend Parasitol 22:416–423
Belli SI, David JP, Ferguson MK, Iveta S, Kelly MJS, Flowers SA, Miska KB, Tomley FM, Shirley MW, Wallach MJ, Smith NC (2009) Conservation of proteins involved in oocyst wall formation in Eimeria maxima, Eimeria tenella and Eimeria acervulina. Int J Parasitol 39:1063–1070
Chapman HD, Roberts B, Shirley MW, Williams RB (2005) Guidelines for evaluating the efficacy and safety of live anticoccidial vaccines, and obtaining approval for their use in chickens and turkeys. Avian Pathol 34:279–290
Crouch CF, Andrews SJ, Ward RG, Francis MJ (2003) Protective efficacy of a live attenuated anticoccidial vaccine administered to 1-day-old chickens. Avian Pathol 32:297–304
Fried M, Mencher D, Sar-Shalom O, Wallach M (1992) Developmental gene expression of a 230-kilodalton macrogamete-specific protein of the avian coccidial parasite, Eimeria maxima. Mol Biochem Parasitol 51:251–262
Hofman FM, Brock M, Taylor CR, Lyons B (1988) IL-4 regulates differentiation and proliferation of human precursor B cells. J Immunol 141:1185–1190
Jang SI, Lillehoj HS, Lee SH, Lee KW, Park MS, Cha SR, Lillehoj EP, Subramanian BM, Sriraman R, Srinivasan VA (2010) Eimeria maxima recombinant Gam82 gametocyte antigen vaccine protects against coccidiosis and augments humoral and cell-mediated immunity. Vaccine 28:2980–2985
Johnson J, Reid WM (1970) Anticoccidial drugs: lesion scoring techniques in battery and floor-pen experiments with chickens. Exp Parasitol 28:30–36
Krücken J, Hosse RJ, Mouafo AN, Entzeroth R, Bierbaum S, Marinovski P, Hain K, Greif G, Wunderlich F (2008) Excystation of Eimeria tenella sporozoites impaired by antibody recognizing gametocyte/oocyst antigens GAM22 and GAM56. Eukaryot Cell 7:202–211
Kumar S, Garg R, Moftah A, Clarke E, MacDonald SE, Chaudhary AS, Sparagano O, Banerjee PS, Kundu K, Tomley FM, Blake DP (2014) An optimized protocol for molecular identification of Eimeria from chickens. Vet Parasitol 199:24–31
Kundu K, Banerjee PS, Garg R, Kumar S, Mandal M, Tomley F, Blake D (2015) Cloning and sequencing of beta-tubulin and internal transcribed spacer-2 (ITS-2) of Eimeria tenella isolate from India. J Parasit Dis 39:539–544
Kundu K, Garg R, Kumar S, Mandal M, Tomley FM, Blake DP, Banerjee PS (2017) Humoral and cytokine response elicited during immunisation with recombinant immune mapped protein-1 (EtIMP-1) and oocysts of Eimeria tenella. Vet Parasitol 244:44–53
Lee SH, Lillehoj HS, Park DW, Jang SI, Morales A, García D, Lucio E, Larios R, Victoria G, Marrufo D, Lillehoj EP (2009) Protective effect of hyperimmune egg yolk IgY antibodies against Eimeria tenella and Eimeria maxima infections. Vet Parasitol 163:123–126
Lillehoj HS, Trout JM (1996) Avian gut-associated lymphoid tissues and intestinal immune responses to Eimeria parasites. Clin Microbiol Rev 9:349–360
Lillehoj HS, Choi KD, Jenkins MC, Vakharia VN, Song KD, Han JY, Lillehoj EP (2000) A recombinant Eimeria protein inducing interferon-γ production: comparison of different gene expression systems and immunization strategies for vaccination against coccidiosis. Avian Dis 44:379–389
Lillehoj HS, Min W, Choi KD, Babu US, Burnside J, Miyamoto T, Rosenthal BM, Lillehoj EP (2001) Molecular, cellular, and functional characterization of chicken cytokines homologous to mammalian IL-15 and IL-2. Vet Immunol Immunopathol 82:229–244
Lillehoj HS, Min W, Dalloul RA (2004) Recent progress on the cytokine regulation of intestinal immune responses to Eimeria. Poult Sci 83:611–623
Lin Z, Shi Y, Deng B, Mao X, Yu D, Li W (2015) Protective immunity against Eimeria tenella infection in chickens following oral immunization with Bacillus subtilis expressing Eimeria tenella 3-1E protein. Parasitol Res 114:3229–3236
Liu D, Cao L, Zhu Y, Deng C, Su S, Xu J, Jin W, Li J, Wu L, Tao J (2014) Cloning and characterization of an Eimeria necatrix gene encoding a gametocyte protein and associated with oocyst wall formation. Parasit Vectors 7:27
Martin AG, Danforth HD, Barta JR, Fernando MA (1997) Analysis of immunological cross protection and sensitivities to anticoccidial drugs among five geographical and temporal strains of E. maxima. Int J Parasitol 5:527–533
Omer FM, Kurtzhals JA, Riley EM (2000) Maintaining the immunological balance in parasitic infections: a role for TGF-beta. Parasitol Today 16:18–23
Peek HW, Landman WJM (2011) Coccidiosis in poultry: anticoccidial products, vaccines and their prevention strategies. Vet Qtly 31:143–161
Rafiqi SI, Garg R, Reena KK, Ram H, Singh M, Banerjee PS (2018) Immune response and protective efficacy of Eimeria tenella recombinant refractile body protein, EtSO7, in chickens. Vet Parasitol 258:108–113
Rose ME, Hesketh P (1976) Immunity to coccidiosis: stages of the life cycle of E. maxima which induce, and are affected by, the response of the host. Parasitology 73:25–37
Song H, Song X, Xu L, Yan R, Shah MA, Li X (2010) Changes of cytokines and IgG antibody in chickens vaccinated with DNA vaccines encoding Eimeria acervulina lactate dehydrogenase. Vet Parasitol 173:219–227
Wallach M (2002) The development of CoxAbic® a novel vaccine against coccidiosis. World Poult 18:24–26
Wallach M, Mencher D, Yarus S, Pillemer G, Halabi A, Pugatsch T (1989) Eimeria maxima: identification of gametocyte protein antigens and their possible role in protective immunity. Exp Parasitol 68:49–56
Wallach M, Halabi A, Pillemer G, Sar-Shalom O, Mencher D, Gilad M, Bendheim U, Danforth HD, Augustine PC (1992) Maternal immunization with gametocyte antigens as a means of providing protective immunity against Eimeria maxima in chickens. Infect Immun 60:2036–2039
Wallach M, Smith NC, Petracca M, Miller CMD, Eckert J, Braun R (1995) Eimeria maxima gametocyte antigens: potential use in a subunit maternal vaccine against coccidiosis in chickens. Vaccine 13:347–354
Wallach MG, Ashash U, Michael A, Smith NC (2003) Field application of a subunit vaccine against an enteric protozoan disease. PLoS One 12:e3948
Williams RB, Andrews SJ (2001) The origins and biological significance of the coccidial lesions that occur in chickens vaccinated with a live attenuated anticoccidial vaccine. Avian Pathol 30:215–220
Xu J, Zhang Y, Tao J (2013) Efficacy of a DNA vaccine carrying Eimeria maxima gam56 antigen gene against coccidiosis in chickens. Korean J Parasitol 2:147–154
Acknowledgements
The authors express sincere thanks to the Director, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, India, for providing the necessary facilities to conduct the study.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
All animal studies were carried out in the Experimental Animal Shed of the Division of Parasitology, ICAR-Indian Veterinary Research Institute, Izatnagar, India. Prior approval for experimental trials and procedures to be used on chickens was obtained from the Institutional Animal Ethics Committee, IVRI (registered with the Committee for the Purpose of Control and Supervision of Experiments on Animals, Ministry of Environment, Forest and Climate Change, Government of India). Permission for the use of recombinant proteins in the experiment was also obtained from the Institutional Biosafety Committee, IVRI, as per the norms of Department of Biotechnology, Government of India.
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Section Editor: David S. Lindsay
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Supplementary Table 1
(DOCX 13 kb)
Supplementary Table 2
(DOCX 13 kb)
Rights and permissions
About this article
Cite this article
Rafiqi, S.I., Garg, R., Ram, H. et al. Immunoprophylactic evaluation of recombinant gametocyte 22 antigen of Eimeria tenella in broiler chickens. Parasitol Res 118, 945–953 (2019). https://doi.org/10.1007/s00436-018-06198-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-018-06198-2