Skip to main content

Advertisement

SpringerLink
Log in

Molecular Characterization and Immunogenicity Analysis of 4D8 Protective Antigen of Hyalomma anatolicum Ticks Collected from Western India

  • Published:
International Journal of Peptide Research and Therapeutics Aims and scope Submit manuscript

Abstract

Vector-borne diseases, mainly transmitted by ticks, have a significant impact on the productivity and health of the animals in the developing countries. The quest for vaccine candidates to control these arthropod vectors remains pivotal and only rationale approach in controlling these diseases. Several candidate antigens have been screened over the years of which 4D8 tick protective antigen have shown promising results in controlling ixodid tick infestations. The 4D8 tick protective antigen is a structural and functional ortholog of insect subolesin/akirin and an evolutionary conserved protein, associated with multiple essential cellular processes in tick biology. Variable efficacy of recombinant subolesin/4D8 vaccine in field trials propels the approach towards synthetic peptide vaccine through reverse vaccinology. Advancement in reverse vaccinology has opened a new paradigm in the search of a perfect candidate antigen. The present study focused on full length amplification, cloning and sequencing of the 4D8 gene from Hyalomma antolicum. Combination of bioinformatics analysis and computational modeling tools identified linear as well as conformational B-cell and MHC class II epitopes conserved in many ixodid ticks which may be useful to design an universal synthetic peptide vaccine against multiple tick species. Western blot analysis using rabbit antisera against recombinant 4D8 protein has also confirmed the immunogenic potential of the candidate.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Almazan C, Kocan KM, Bergman DK, Garcia-Garcia JC, Blouin EF, de la Fuente J (2003) Identification of protective antigens for the control of Ixodes scapularis infestations using cDNA expression library immunization. Vaccine 21:1492–1501

    Article  CAS  PubMed  Google Scholar 

  • Almazan C, Lagunes R, Villar M, Canales M, Rosario-Cruz R, Jongejan F, de laFuente J (2010) Identification and characterization of Rhipicephalus (Boophilus) microplus candidate protective antigens for the control of cattle tick infestations. Parasitol Res 106:471–479

    Article  PubMed  Google Scholar 

  • Almazan C, Moreno-Cantu O, Moreno-Cid JA, Galindo RC, Canales M, Villar M, de la Fuente J (2012) Control of tick infestations in cattle vaccinated with bacterial membranes containing surface-exposed tick protective antigens. Vaccine 30:265–272

    Article  CAS  PubMed  Google Scholar 

  • Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Benkert P, Kunzli M, Schwede T (2009) QMEAN server for protein model quality estimation. Nucleic Acids Res 37:W510–W514

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Blom N, Gammeltoft S, Brunak S (1999) Sequence- and structure-based prediction of eukaryotic protein phosphorylation sites. J Mol Biol 294:1351–1362

    Article  CAS  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1–2):248–254

    Article  CAS  PubMed  Google Scholar 

  • Canales M, Enriquez A, Ramos E, Cabrera D, Dandie H, Soto A, Falcon V, Rodriguez M, de la Fuente J (1997) Large-scale production in Pichia pastoris of the recombinant vaccine Gavac against cattle tick. Vaccine 15:414–422

    Article  CAS  PubMed  Google Scholar 

  • Canales M, Naranjo V, Almazan C, Molina R, Tsuruta SA, Szabo MP, Manzano-Roman R, Perez de la Lastra JM, Kocan KM, Jimenez MI, Lucientes J, Villar M, de la Fuente J (2009a) Conservation and immunogenicity of the mosquito ortholog of the tick-protective antigen,subolesin. Parasitol Res 105:97–111

    Article  PubMed  Google Scholar 

  • Canales M, Ballesteros C, Moreno-Cid JA, Espinosa AM, Villar M, de la Fuente J (2009b) Extractive bioconversion to produce the Aedes albopictus akirin in an aqueous two-phase system supporting Pichia pastoris growth and protein secretion. Biochem Eng J 46:105–114

    Article  CAS  Google Scholar 

  • Carpio Y, Basabe L, Acosta J, Rodríguez A, Mendoza A, Lisperger A, Zamorano E, González M, Rivas M, Contreras S, Haussmann D, Figueroa J, Osorio VN, Asencio G, Mancilla J, Ritchie G, Borroto C, Estrada MP (2011) Novel gene isolated from Caligus rogercresseyi: a promising target for vaccine development against sea lice. Vaccine 29:2810–2820

    Article  CAS  PubMed  Google Scholar 

  • De Castro E, Sigrist CJA, Gattiker A, Bulliard V, Langendijk-Genevaux PS, Gasteiger E, Bairoch A, Hulo N (2006) ScanProsite: detection of PROSITE signature matches and ProRule-associated functional and structural residues in proteins. Nucleic Acids Res 34:W362–W365

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • de la Fuente J, Kocan KM (2003) Advances in the identification and characterization of protective antigens for development of recombinant vaccines against tick infestations. Expert Rev Vaccines 2:583–593

    Article  PubMed  Google Scholar 

  • de la Fuente J, Merrino O (2013) Vaccinomics, the new road to tick vaccines. Vaccine 31:5923–5929

    Article  CAS  PubMed  Google Scholar 

  • de la Fuente J, Rodriguez M, Redondo M, Montero C, Garcia-Garcia JC, Mendez L, Serrano E, Valdes M, Enriquez A, Canales M, Ramos E, Boue O, Machado H, Lleonart R, de Armas CA, Rey S, Rodriguez JL, Artiles M, Garcia L (1998) Field studies and cost-effectiveness analysis of vaccination with Gavac™ against cattle tick Boophilus microplus. Vaccine 16:366–373

    Article  PubMed  Google Scholar 

  • de la Fuente J, Almazan C, Canales M, Pérez de la Lastra JM, Kocan KM, Willadsen P (2007) A ten-year review of commercial vaccine performance for control of tick infestations on cattle. Anim Health Res Rev 8:23–28

    Article  PubMed  Google Scholar 

  • FAO (2004) Resistance management and integrated parasite control in ruminants, Guidelines, Animal Production and Health Division, FAO, p. 25–77

  • Feng ZP, Zhang X, Han P, Arora N, Anders RF, Norton RS (2006) Abundance of intrinsically unstructured proteins in P. falciparum and other apicomplexan parasite proteomes. Mol Biochem Parasitol 150:256–267

    Article  CAS  PubMed  Google Scholar 

  • Galindo RC, Doncel-Perez E, Zivkovic Z, Naranjo V, Gortazar C, Mangold AJ, Martin-Hernando MP, Kocan KM, de la Fuente J (2009) Tick subolesin is an ortholog of the akirins described in insects and vertebrates. Dev Comp Immunol 33:612–617

    Article  CAS  PubMed  Google Scholar 

  • Ghosh S, Bansal GC, Gupta SC, Ray DD, Khan QM, Irshad H, Shahiduzzaman Md, Seitzer U, Ahmed JS (2007) Status of tick distribution in Bangladesh, India and Pakistan. Parasitol Res 101:207–216

    Article  Google Scholar 

  • Goto A, Matsushita K, Gesellchen V, El Chamy L, Kuttenkeuler D, Takeuchi O, Hoffmann JA, Akira S, Boutros M, Reichhart JM (2008) Akirins are highly conserved nuclear proteins required for NF-kappa B-dependent gene expression in drosophila and mice. Nat Immunol 9:97–104

    Article  CAS  PubMed  Google Scholar 

  • Graf JF, Gogolewski R, Leach-Bing N, Sabatini GA, Molento MB, Bordin EL, Arantes GJ (2004) Tick control: an industry point of view. Parasitol 129:S427–S442

    Article  Google Scholar 

  • Guerrero FD, Miller RJ, Pérez de León AA (2012) Cattle tick vaccines: many candidate antigens, but will a commercially viable product emerge? Int J Parasitol 42:421–427

    Article  CAS  PubMed  Google Scholar 

  • Gupta R, Jung E, Brunak S (2004) Prediction of N-glycosylation sites in human proteins. http://www.cbs.dtu.dk/services/NetNGlyc/. (In preparation)

  • Hajdusek O, Sojka D, Kopacek P, Buresova V, Franta Z, Sauman I, Winzerling J, Grubhoffer L (2009) Knockdown of proteins involved in iron metabolism limits tick reproduction and development. Proc Natl Acad Sci USA 106:1033–1038

    Article  PubMed  PubMed Central  Google Scholar 

  • Hajdusek O, Almazan C, Loosova G, Villar M, Canales M, Grubhoffer L, Kopacek P, de la Fuente J (2010) Characterization of ferritin 2 for the control of tick infestations. Vaccine 28:2993–2998

    Article  CAS  PubMed  Google Scholar 

  • Harrington D, Canales M, de la Fuente J, de Luna C, Robinson K, Guy J, Sparagano O (2009) Immunisation with recombinant proteins subolesin and Bm86 for the control of Dermanyssus gallinae in poultry. Vaccine 27:4056–4063

    Article  CAS  PubMed  Google Scholar 

  • Jones DT (1999) Protein secondary structure prediction based on position-specific scoring matrices. J Mol Biol 292:195–202

    Article  CAS  PubMed  Google Scholar 

  • Jongejan F, Uilenberg G (2004) The global importance of ticks. Parasitology 129:S3–S14

    Article  PubMed  Google Scholar 

  • Källberg M, Wang H, Wang S, Peng J, Wang Z, Lu H, Xu J (2012) Template-based protein structure modeling using the RaptorX web server. Nat Protoc 7:1511–1522

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kelley LA, Sternberg MJE (2009) Protein structure prediction on the web: a case study using the Phyre server. Nat Protoc 4:363–371

    Article  CAS  PubMed  Google Scholar 

  • Kiefer F, Arnold K, Künzli M, Bordoli L, Schwede T (2009) The SWISS-MODEL repository and associated resources. Nucleic Acids Res 37:D387–D392

    Article  CAS  PubMed  Google Scholar 

  • Kolaskar AS, Tongaonkar PC (1990) A semi-empirical method for prediction of antigenic determinants on protein antigens. FEBS Lett 276:172–174

    Article  CAS  PubMed  Google Scholar 

  • Kopp J, Schwede T (2006) The SWISS-MODEL repository: new features and functionalities. Nucleic Acids Res 34:D315–D318

    Article  CAS  PubMed  Google Scholar 

  • Liu ZQ, Xia J, Wang GL, Kuermanali N (2016) Cloning and expression of the 4D8 gene from Hyalomma asiaticum tick. Genet Mol Res 15(2):gmr.15027951. https://doi.org/10.4238/gmr.15027951

    Article  CAS  Google Scholar 

  • Manzano-Roman R, Diaz-Martin V, Oleaga A, Siles-Lucas M, Perez-Sanchez R (2012) Subolesin/akirin orthologs from Ornithodoros spp. soft ticks: cloning RNAi gene silencing and protective effect of the recombinant proteins. Vet Parasitol 185:248–259

    Article  CAS  PubMed  Google Scholar 

  • Merino O, Almazán C, Canales M, Villar M, Moreno-Cid JA, Estrada-Pena A, Kocan KM, de la Fuente J (2011) Control of Rhipicephalus (Boophilus) microplus infestations by the combination of subolesin vaccination and tick autocidal control after subolesin gene knockdown in ticks fed on cattle. Vaccine 29:2248–2254

    Article  CAS  PubMed  Google Scholar 

  • Minjauw L, McLeod A (2003) Tick borne diseases and poverty. The impact of tick and tick borne diseases on the livelihoods of small-scale and marginal livestock owners in India and eastern and southern Africa. In: Research report. DFID animal health programme. Centre for Tropical Veterinary Medicine, University of Edinburgh, UK

    Google Scholar 

  • Moreno-Cid JA, Jimenez M, Cornelie S, Molina R, Alarcon P, Lacroix MN, Pinal R, Delacour S, Lucientes J, Canales M, Perez de la Lastra JM, Villar M, de la Fuente J (2010) Characterization of Aedes albopictus akirin for the control of mosquito and sand fly infestations. Vaccine 29:77–82

    Article  CAS  PubMed  Google Scholar 

  • Moreno-Cid JA, Perez de la Lastra JM, Villar M, Jimenez M, Pinal R, Estrada-Pena A, Molina R, Lucientes J, Gortazar C, de la Fuente J (2013) Control of multiple arthropod vector infestations with subolesin/akirin vaccines. Vaccine 31:1187–1196

    Article  CAS  PubMed  Google Scholar 

  • Nielsen M, Lund O (2009) NN-align. An artificial neural network-based alignment algorithm for MHC class II peptide binding prediction. BMC Bioinformatics 10:296

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nielsen M, Lundegaard C, Lund O (2007) Prediction of MHC class II binding affinity using SMM-align, a novel stabilization matrix alignment method. BMC Bioinform 8:238

    Article  CAS  Google Scholar 

  • Nuttall PA, Trimnell AR, Kazimirova M, Labuda M (2006) Exposed and concealed antigens as vaccine targets for controlling ticks and tick-borne diseases. Parasite Immunol 28:155–163

    Article  CAS  PubMed  Google Scholar 

  • Petersen TN, Brunak S, von Heijne G, Nielsen H (2011) SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 8:785–786

    Article  CAS  PubMed  Google Scholar 

  • Ponomarenko JV, Bui H, Li W, Fusseder N, Bourne PE, Sette A, Peters B (2008) ElliPro: a new structure-based tool for the prediction of antibody epitopes. BMC Bioinform 9:514

    Article  CAS  Google Scholar 

  • Prudencio CR, Perez de la Lastra JM, Canales M, Villar M, de la Fuente J (2010) Mapping protective epitopes in the tick and mosquito subolesin ortholog proteins. Vaccine 28:5398–5406

    Article  CAS  PubMed  Google Scholar 

  • Rachinsky A, Guerrero FD, Scoles GA (2008) Proteomic profiling of Rhipicephalus (Boophilus) microplus midgut responses to infection with Babesia bovis. Vet Parasitol 152:294–313

    Article  CAS  PubMed  Google Scholar 

  • Sajid MS, Iqbal Z, Khan MN, Muhammad G, Iqbal MU (2007) Effect of Hyalomma ticks (Acari: Ixodidae) on milk production of dairy buffaloes (Bos bubalus bubalis) of Punjab (Pakistan). Ital J Anim Sci 6:939–941

    Article  Google Scholar 

  • Shakya M, Kumar B, Nagar G, de la Fuente J, Ghosh S (2014) Subolesin: a candidate vaccine antigen for the control of cattle tick infestations in Indian situation. Vaccine 32:3488–3494

    Article  CAS  PubMed  Google Scholar 

  • Sharma AK, Kumar R, Kumar S, Nagar G, Singh NK, Rawat SS, Dhakadd ML, Rawat AKS, Ray DD, Ghosh S (2012) Deltamethrin and cypermethrin resistance status of Rhipicephalus (Boophilus) microplus collected from six agroclimatic regions of India. Vet Parasitol 188:337–345

    Article  CAS  PubMed  Google Scholar 

  • Singh NK, Haque M, Singh H, Rath SS, Ghosh S (2014) A comparative study on cypermethrin resistance in Rhipicephalus (Boophilus) microplus and Hyalomma anatolicum from Punjab (India). Ticks Tick Borne Dis 5:90–94

    Article  PubMed  Google Scholar 

  • Steentoft C, Vakhrushev SY, Joshi HJ, Kong Y, Vester-Christensen MB, Schjoldager KT, Lavrsen K, Dabelsteen S, Pedersen NB, Marcos-Silva L, Gupta R, Bennett EP, Mandel U, Brunak S, Wandal HH, Levery SB, Clausen H (2013) Precision mapping of the human O-GalNAc glycoproteome through Simple Cell technology. EMBO J 32:1478–1488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sugase K, Dyson HJ, Wright PE (2007) Mechanism of coupled folding and binding of an intrinsically disordered protein. Nature 447:1021–1025

    Article  CAS  PubMed  Google Scholar 

  • Sun J, Wu D, Xu T, Wang X, Xu X, Tao L, Li YX, Cao ZW (2009) SEPPA: a computational server for spatial epitope prediction of protein antigens. Nucleic Acids Res 37:W612–W616

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sutherst RW (1983) Management of Arthropod Parasitism in Livestock. World Association for the Advancement of Veterinary Parasitology, Dansmore

    Google Scholar 

  • Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. https://doi.org/10.1093/molbev/msr121

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang P, Sidney J, Dow C, Mothe B, Sette A, Peter B (2008) A systematic assessment of MHC class II peptide binding predictions and evaluation of a consensus approach. PLoS Comput Biol 4:e1000048

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ward JJ, Sodhi JS, McGuffin LJ, Buxton BF, Jones DT (2004) Prediction and functional analysis of native disorder in proteins from the three kingdoms of life. J Mol Biol 337:635–645

    Article  CAS  PubMed  Google Scholar 

  • Willadsen P (1997) Novel vaccines for ectoparasites. Vet Parasitol 71:209–212

    Article  CAS  PubMed  Google Scholar 

  • Willadsen P, Bird P, Cobon GS, Hungerford J (1995) Commercialisation of a recombinant vaccine against Boophilus microplus. Parasitology 110:S43–S50

    Article  PubMed  Google Scholar 

  • Zhang Y (2007) Template-based modeling and free modeling by I-TASSER in CASP7. Proteins 69:108–117

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y (2008) I-TASSER server for protein 3D structure prediction. BMC Bioinform 9:40

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are thankful to the laboratory facilities extended by the authorities of Central Institute of Research on Buffaloes, Hissar, Haryana, India. The first author is thankful to the Department of Science and Technology, Government of India for the PhD program.

Author information

Authors and Affiliations

  1. Department of Veterinary Physiology and Biochemistry, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, 125004, India

    Mayukh Ghosh & N. Sangwan

  2. Division of Biological Products, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, 243122, India

    S. Chakravarti

  3. Department of Veterinary Microbiology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, 125004, India

    Somesh Banerjee

  4. Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, 125004, India

    A. Ghosh

  5. Department of Veterinary Physiology, Veterinary College, Pookode, Lakkidi, Kerala, 673576, India

    R. Kumar

  6. Department of Veterinary Parasitology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, 125004, India

    A. K. Sangwan

Authors
  1. Mayukh Ghosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. N. Sangwan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. Chakravarti
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Somesh Banerjee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. Ghosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. R. Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A. K. Sangwan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mayukh Ghosh.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ghosh, M., Sangwan, N., Chakravarti, S. et al. Molecular Characterization and Immunogenicity Analysis of 4D8 Protective Antigen of Hyalomma anatolicum Ticks Collected from Western India. Int J Pept Res Ther 25, 1291–1308 (2019). https://doi.org/10.1007/s10989-018-9776-8

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10989-018-9776-8

Keywords

Search

Navigation