Veterinary Research Forum
Vet Res Forum

Introducing a new anti-Rhipicephalus (Boophilus) microplus tick recombinant vaccine candidate using cathepsin and tropomyosin multi-epitope gene

Document Type : Original Article

Authors

Zahra Asadollahi 1
Sedigheh Nabian 1, 2
Mohammad Taheri 3
Elaheh Ebrahimzadeh 4

1 Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Center of Tick and Tick-borne Disease, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Rastegar Reference Laboratory, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

4 Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

https://doi.org/10.30466/vrf.2021.113699.2704
Abstract
Rhipicephalus (Boophilus) spp. are important vectors for Babesia and Anaplasma species causing severe economic losses in livestock. Chemical compounds are commonly used to control tick infestation; however, acaricides resistance in tick has led to move toward alternative strategies such as vaccination. In this study, we introduced a vaccine candidate, namely CaTro against Rh. microplus tick composing of immunogenic B-cell epitopes derived from Rh. microplus cathepsin L and tropomyosin proteins. To evaluate this vaccine candidate, firstly the CaTro sequence was inserted into the prokaryotic expression vector and the recombinant protein CaTro was expressed in Bl21 bacteria. Afterward, purification was performed by Ni-NTA affinity chromatography. The quality of purified recombinant CaTro was also analyzed using sodium dodecyl sulfate-gel electrophoresis and western blotting. Moreover, to evaluate the induction of immune response, the rabbits were immunized with purified recombinant protein combined with Freund’s adjuvant. The findings of this study revealed molecular weight of expressed protein (CaTro) as 38.00 kDa. Furthermore, anti-CaTro antibody was detected in immunized rabbit's sera through dot blotting; while, there was not any response to the control rabbit's sera. The results suggest that CaTro is a potential candidate to develop an anti- Rh. microplus tick.

Keywords

Cathepsin
Epitope
Tropomyosin
Vaccine
  1.  Vilela VLR, Feitosa TF, Bezerra RA, et al. Multiple acaricide-resistant Rhipicephalus microplus in the semi-arid region of Paraíba State, Brazil. Ticks Tick Borne Dis 2020; 11(4): 101413. doi: 10.1016/j.ttbdis. 2020.101413. 
  2. El-Ashram S, Aboelhadid SM, Kamel AA, et al. First report of cattle tick Rhipicephalus (Boophilus) annulatus in Egypt resistant to ivermectin. Insects 2019; 10(11): 404. doi: 10.3390/insects10110404.
  3. Akhtar M, Muhammad F, Lodhi LA, et al. Immunity against ticks- a review. Pak Vet J 2011; 31(1): 9-16.
  4. Popara M, Villar M, Mateos-Hernández L, et al. Lesser protein degradation machinery correlates with higher BM86 tick vaccine efficacy in Rhipicephalus annulatus when compared to Rhipicephalus microplus. Vaccine 2013; 31(42): 4728-4735.
  5. Canales M, Almazán C, Naranjo V, et al. Vaccination with recombinant Boophilus annulatus Bm86 ortholog protein, Ba86, protects cattle against annulatus and B. microplus infestations. BMC Biotechnol 2009; 9:29. doi:10.1186/1472-6750-9-29.
  6. Parizi LF, Githaka NW, Logullo C, et al. The quest for universal vaccine against ticks: cross-immunity insights. Vet J 2012; 194(2): 158-165.
  7. Xavier MA, Tirloni L, Torquato R, et al. Blood anticlotting activity of Rhipicephalus microplus cathepsin L-like enzyme. Biochimie 2019; 163:12-20.
  8. Taylor MJ, Jenkins RE, Bianco AE. Protective immunity induced by vaccination with Onchocerca volvulus tropomyosin in rodents. Parasite Immunol 1996; 18(5): 219-225.
  9. Wright HW, Bartley K, Huntley JF, et al. Characterization of tropomyosin and paramyosin as vaccine candidate molecules for the poultry red mite, Dermanyssus gallinae. Parasites Vectors 2016; 9.544. doi: 10.1186/s13071-016-1831-8.
  10. Nabian S, Taheri M, Mazaheri Nezhad Fard R, et al. Identification of tropomyosin and its immunological properties from larvae of cattle tick, Boophilus annulatus. Iran J Parasitol 2013; 8(2): 242-248.
  11. Lehman W, Craig R, Vibert P. Ca2+-Induced tropomyosin movement in Limulus thin filaments revealed by three-dimensional reconstruction. Nature 1994; 368: 65-67.
  12. Wang CL, Coluccio LM. New insights into the regulation of the actin cytoskeleton by tropomyosin. Int Rev Cell Mol Biol 2010; 281: 91-128.
  13. Hajighahramani N, Nezafat N, Eslami M, et al. Immunoinformatics analysis and in silico designing of a novel multi-epitope peptide vaccine against Staphylococcus aureus. Infect Genet Evol. 2017; 48: 83-94.
  14. Cao A, Liu Y, Wang J, et al. 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 2015; 33(48):6757-6762.
  15. Martin DM, Berriman M, Barton GJ. GOtcha: a new method for prediction of protein function assessed by the annotation of seven genomes. BMC Bioinform 2004; 5, 178. doi: 10.1186/1471-2105-5-178.
  16. Bai Y, He S, Zhao G, et al. Toxoplasma gondii: bioinformatics analysis, cloning and expression of a novel protein TgIMP1. Exp. Parasitol 2012; 132(4): 458-464.
  17. Changklungmoa N, Poinok N, Yencham C, et al. Vaccine potential of recombinant cathepsinL1G against Fasciola gigantica in mice. Vet Parasitol 2016;226: 124-131.
  18. Renard G, Garcia JF, Cardoso FC, et al. Cloning and functional expression of a Boophilus microplus cathepsin L-like enzyme. Insect Biochem Mol Biol 2000; 30(11): 1017-1026.
  19. Clara RO, Soares TS, Torquato RJS, et al. Boophilus microplus cathepsin L-like (BmCL1) cysteine protease: Specificity study using a peptide phage display library. Vet Parasitol 2011; 181(2-4): 291-300.
  20. Asadollahi A, Nabian S, Taheri M, et al. Designing of potential vaccine candidates of fused cathepsin L and tropomyosin genes of Rhipicephalus (Boophilus) annulatus tick larva. J Bas Res Med Sci 2018; 5(3): 9-16.
  21. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227(5259): 680-685.
  22. Warburg O, Christian W. Protein estimation by extinction at 260 nm and 280 nm. Biochem Z 1941; 310: 384-392.
  23. Willadsen P. Tick control: thoughts on a research agenda. Vet Parasitol 2006; 138(1-2): 161-168.
  24. Cobon G, Hungerford J, Woodrow M, et al. Vaccination against Boophilus microplus: the Australian field experience. In proceedings: Recombinant vaccines for the control of cattle tick. La Habana, Cuba 1995; 163-175.
  25. García-García JC, Gonzalez IL, González DM, et al. Sequence variations in the Boophilus microplus Bm86 locus and implications for immunoprotection in cattle vaccinated with this antigen, Exp Appl Acarol 1991; 23(11):883-895.
  26. Ramírez Rodríguez PB, Cruz RR, Domínguez García DI, et al. Identification of immunogenic proteins from ovarian tissue and recognized in larval extracts of Rhipicephalus (Boophilus) microplus, through an immunoproteomic approach. Exp Parasitol 2016; 170: 227-235.
  27. Imamura S, Konnai S, Vaz Ida S, et al. Effects of anti-tick cocktail vaccine against Rhipicephalus appendiculatus. Jpn J Vet Res 2008; 56(2): 85-98.
  28. Aguirre Ade A, Lobo FP, Cunha RC, et al. Design of the ATAQ peptide and its evaluation as an immunogen to develop a Rhipicephalus Vet Parasitol 2016; 221: 30-38.
  29. Sojka D, Francischetti IMB, Calvo E, et al. Cysteine proteases from blood feeding arthropod ectoparasites. Adv Exp Med Biol 2011; 712: 177-191.
  30. Zhao G, Zhou A, Lv G, et al. Toxoplasma gondii cathepsin proteases are undeveloped prominent vaccine antigens against toxoplasmosis. BMC Infect Dis 2013; 13: 207. doi: 10.1186/1471-2334-13-207.
  31. Wesołowska A, Basałaj K, Norbury LJ, et al. Vaccination against Fasciola hepatica using cathepsin L3 and B3 proteases delivered alone or in combination. Vet Parasitol 2018; 250: 15-21.
  32. Saidi S, Nabian S, Ebrahimzade E, et al. Identification and characterization of a cathepsin L-like cysteine protease from Rhipicephalus (Boophilus) annulatus. Exp Appl Acarol 2016; 68(2): 251-265.
  33. Huntley JF, Machell J, Nisbet AJ, et al. Identification of tropomyosin, paramyosin and apolipophorin/vitello-genin as three major allergens of the sheep scab mite, Psoroptes ovis. Parasite Immunol 2004; 26(8-9): 335-342.
  34. Ranjbar MM, Gupta SH, Ghorban KH, et al. Designing and modeling of complex DNA vaccine based on tropomyosin protein of Boophilus genus tick. Appl Biochem Biotechnol 2015;175(1):323-339.
  35. Baig F, Fernando LP, Salazar MA, et al. Dynamic transcriptional response of Escherichia coli to inclusion body formation. Biotechnol Bioeng 2014; 111(5): 980-999.
  36. Parizi LF, Reck J Jr, Oldiges DP, et al. Multi-antigenic vaccine against the cattle tick Rhipicephalus (Boophilus) microplus: a field evaluation. Vaccine 2012; 30(48); 6912-6917.
  37. Ahmad Fuaad AA, Roubille R, Pearson MS, et al. The use of a conformational cathepsin D-derived epitope for vaccine development against Schistosoma mansoni. Bioorg Med Chem 2015; 23(6):1307-1312.
  38. You MJ. Immunization of mice with recombinant P27/30 protein confers protection against hard tick Haemaphysalis longicornis (Acari: Ixodidae) infestation. J Vet Sci 2005; 6(1): 47-51.
  39. Leal AT, Seixas A, Pohl PC, et al. Vaccination of bovines with recombinant Boophilus Yolk pro-Cathepsin. Vet Immunol Immunopathol 2006; 114(3-4); 341-345.
  40. Chantree P, Phatsara M, Meemon K, et al. Vaccine potential of recombinant cathepsin B against Fasciola gigantica. Exp Parasitol 2013; 135(1):102-109.
  41. Wabl M, Steinberg C. Affinity maturation and class switching. Curr Opin Immunol 1996; 8(1): 89-92.
Veterinary Research Forum
Volume 12, Issue 4
Autumn 2021
Pages 445-450

  • Receive Date 02 September 2019
  • Revise Date 18 April 2021
  • Accept Date 08 May 2021

Home

Guide for Authors

Submit Manuscript

Reviewers

Contact Us