Development and Evaluation of an Δ aroA / Δ htrA Salmonella enteritidis Vector Expressing Eimeria maxima TRAP Family Protein EmTFP250 with CD 154 (CD 40L) as Candidate Vaccines against Coccidiosis in Broilers
DOI:
https://doi.org/10.3923/ijps.2010.1031.1037Keywords:
Apicomplexa, coccidiosis, eimeria, Salmonella, TRAP, vaccinationAbstract
Coccidiosis is caused by parasites of the genus Eimeria, belonging to phylum Apicomplexa. EmTFP250 is a high molecular mass, asexual stage antigen from Eimeria maxima (EM) strongly associated with maternal immunity in newly hatched chickens. Cloning and sequence analysis predict the antigen to be a novel member of the Thrombospondin-Related Adhesive Protein (TRAP) family. Three novel attenuated Salmonella enteritidis strains (ΔSE) expressing TRAP oligopeptides in association with a potential immune-enhancing CD 154 sequence, on the outer membrane protein lamB, were developed. Broiler chicks were grouped based on treatment and 108 cfu/chick of vectors expressing one of three sequences, or vehicle alone, was orally administered to each group. At 21 d of age, all groups were challenged with 104 sporulated oocysts/chick orally. Mortality at 5d post-challenge was markedly different (p<0.05) in chickens vaccinated with TRAP Upstream (US). To further evaluate the efficacy of TRAP US as a potential vaccine candidate, a similar study was conducted. Broilers were orally vaccinated with 108 cfu/chick vehicle with TRAP US and CD 154 or sham vaccinated with saline. Coccidia challenge was performed with 105 sporulated oocysts/chick at 22 d of age. Immunized chickens showed remarkable improvement in weight gain (p<0.05) and had reduced mortality (p = 0.055) when compared to non-immunized controls. These two studies underscore the potential of EmTFP250 as a potential candidate for a recombinant vaccine targeting coccidiosis in chickens.
References
Allen, P.C. and R.H. Fetterer, 2002. Recent advances in biology and immunobiology of Eimeria species and in diagnosis and control of infection with these coccidian parasites of poultry. Clin. Microbiol. Rev., 15: 58-65.
Ashby, D., I. Leduc, W. Lauzon, B. Craig Lee, N. Singhal and D.W. Cameron, 2005. Attenuated Salmonella typhimurium SL3261 as a vaccine vector for recombinant antigen in rabbits. J. Immunol. Methods, 299: 153-164.
Blake, D.P., M.W. Shirley and A.L. Smith, 2006. Genetic identification of antigens protective against coccidia. Parasite Immunol., 28: 305-314.
Bumstead, J., P. Dunn and F. Tomley, 1995. Nitrocellulose immunoblotting for identification and molecular gene cloning of Eimeria maxima antigens that stimulate lymphocyte proliferation. Clin. Diagn. Lab. Immunol., 2: 524-530.
Burns, D.M. and I.R. Beacham, 1985. Rare codons in E. coli and S. typhimurium signal sequences. FEBS Lett., 189: 318-324.
Carruthers, V. and J.C. Boothroyd, 2007. Pulling together: An integrated model of Toxoplasma cell invasion. Curr. Opin. Microbiol., 10: 83-89.
Cheeseman, J.H., N.A. Levy, P. Kaiser, H.S. Lillehoj and S.J. Lamont, 2008. Salmonella enteritidis-induced alteration of inflammatory CXCL-chemokine Messenger-RNA gene expression and histologic changes in the ceca of infected chicks. Avian Dis., 25: 229-234.
Cox, M., S. Layton, T. Jiang, K. Cole and B. Hargis et al., 2007. Scarless and site-directed mutagenesis in Salmonella enteritidis chromosome. BMC Biotechnol., 7: 59-59.
Escalante, A., E. Barrio and F. Ayala, 1995. Evolutionary origin of human and primate malarias: Evidence from the circumsporozoite protein gene. Mol. Biol. Evol., 12: 616-626.
Hong, Y.H., H.S. Lillehoj, S.H. Lee, R.A. Dalloul and E.P. Lillehoj, 2006. Analysis of chicken cytokine and chemokine gene expression following Eimeria acervulina and Eimeria tenella infections. Vet. Immunol. Immunopathol., 114: 209-223.
Husseiny, M.I. and M. Hensel, 2005. Rapid method for the construction of Salmonella enterica Serovar Typhimurium vaccine carrier strains. Infect. Immun., 73: 1598-1605.
Johnson, J. and W.M. Reid, 1970. Anticoccidial drugs: Lesion scoring techniques in battery and floor-pen experiments with chickens. Exp. Parasitol., 28: 30-36.
Kang, Y., T. Durfee, J.D. Glasner, Y. Qiu, D. Frisch, K.M. Winterberg and F.R. Blattner, 2004. Systematic mutagenesis of the Escherichia coli genome. J. Bacteriol., 186: 4921-4930.
Konjufca, V., M. Jenkins, S. Wang, M.D. Juarez-Rodriguez and R. Curtiss III, 2008. Immunogenicity of recombinant attenuated Salmonella enterica serovar typhimurium vaccine strains carrying a gene that encodes Eimeria tenella antigen SO7. Infect. Immun., 76: 5745-5753.
Konjufca, V., S. Wanda, M.C. Jenkins and R. Curtiss III, 2006. A recombinant attenuated Salmonella enterica Serovar Typhimurium vaccine encoding Eimeria acervulina antigen offers protection against E. acervulina challenge. Infect. Immun., 74: 6785-6796.
Layton, S.L., D.R. Kapczynski, S. Higgins, J. Higgins and A.D. Wolfenden et al., 2009. Vaccination of chickens with recombinant Salmonella expressing M2e and CD154 epitopes increases protection and decreases viral shedding after low pathogenic avian influenza challenge. Poult. Sci., 88: 2244-2252.
Li, L., J. Crabtree, S. Fischer, D. Pinney, C.J. Stoeckert Jr., L.D. Sibley and D.S. Roos, 2004. ApiEST-DB: Analyzing clustered EST data of the apicomplexan parasites. Nucl. Acids Res., 32: D326-D328.
Lillehoj, H.S., W. Min and R.A. Dalloul, 2004. Recent progress on the cytokine regulation of intestinal immune responses to Eimeria. Poult. Sci., 83: 611-623.
Long, P.L., B.J. Millard, L.P. Joyner and C.C. Norton, 1976. A guide to laboratory techniques used in the study and diagnosis of avian coccidiosis. Folia Vet. Lat., 6: 201-217.
Martin, A.G., H.D. Danforth, J.R. Barta and M.A. Fernando, 1997. Analysis of immunological cross-protection and sensitivities to anticoccidial drugs among five geographical and temporal strains of Eimeria maxima. Intl. J. Parasitol., 27: 527-533.
O'Meara, K.M., C.J. Kremer, S.L. Layton, L.R. Berghman, B.M. Hargis and K.M. Cole, 2010. Evaluation of recombinant Salmonella expressing CD154 for persistence and enhanced antibody response in commercial Turkeys. Poult. Sci., 89: 1399-1405.
Quezada, S.A., L.Z. Jarvinen, E.F. Lind and R.J. Noelle, 2004. CD40/CD154 interactions at the interface of tolerance and immunity. Annu. Rev. Immunol., 22: 307-328.
Shirley, M.W., A.L. Smith and D.P. Blake, 2007. Challenges in the successful control of the avian coccidia. Vaccine, 25: 5540-5547.
Smith, N.C., M. Wallach, C.M. Miller, R. Morgenstern, R. Braun and J. Eckert, 1994. Maternal transmission of immunity to Eimeria maxima: Enzyme linked immunosorbant assay analysis of protective antibodies induced by infection. Infect. Immunol., 62: 1348-1357.
Song, K.D., H.S. Lillehoj, K.D. Choi, C.H. Yun, M.S. Parcells, J.T. Huynh and J.Y. Han, 2000. A DNA vaccine encoding a conserved Eimeria protein induces protective immunity against live Eimeria acervulina challenge. Vaccine, 19: 243-252.
Spaccapelo, R., S. Naitza, K.J. Robson and A. Crisanti, 1997. Thrombospondin-Related Adhesive Protein (TRAP) of plasmodium berghei and parasite motility. The Lancet, 350: 335-335.
Spreng, S., G. Dietrich and G. Weidinger, 2006. Rational design of Salmonella-based vaccination strategies. Methods, 38: 133-143.
Talebi, A. and G. Mulcahy, 1994. High-resolution mapping of B-cell epitopes within an antigenic sequence from Eimeria tenella. Infect. Immun., 62: 4202-4207.
Tomley, F.M., K.J. Billington, J.M. Bumstead, J.D. Clark and P. Monaghan, 2001. EtMIC4: A microneme protein from Eimeria tenella that contains tandem arrays of epidermal growth factor-like repeats and thrombospondin type-I repeats. Int. J. Parasitol., 31: 1303-1310.
Williams, R.B. and J. Catchpole, 2000. A new protocol for a challenge test to assess the efficacy of live anticoccidial vaccines for chickens. Vaccine, 18: 1178-1185.
Witcombe, D.M., S.I. Belli, M.G. Wallach and N.C. Smith, 2003. Molecular characterisation of EmTFP250: A novel member of the TRAP protein family in Eimeria maxima. Int. J. Parasitol., 33: 691-702.
Xu, H., G. Zhao, X. Huang, Z. Ding and J. Wang et al., 2010. CD40-expressing plasmid induces anti-CD40 antibody and enhances immune responses to DNA vaccination. J. Gene Med., 12: 97-106.
Yun, C.H., H.S. Lillehoj and E.P. Lillehoj, 2000. Intestinal immune responses to coccidiosis. Dev. Comp. Immunol., 24: 303-324.
Zar, J.H., 1984. Biostatistical Analysis. 2nd Edn., Prentice-Hall Inc., Englewood Cliffs, New Jersey, USA, Pages: 718.
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