The chemical conjugate pP0-Bm86 as an antigen of a broad-spectrum vaccine against ticks
Keywords:
péptido P٠, conjugado químico, vacuna, garrapatasAbstract
Introduction. Ticks are ectoparasites, which are vectors of infectious diseases for humans and animals. Vaccination could be an ecological alternative to chemicals to control these ectoparasites. A peptide of the ribosomal protein P0 conjugated to hemocyanin from the mollusk Megathura crenulata (KLH) has shown an effectiveness of more than 90 % against Rhipicephalus sanguineus and R. microplus tick species. However, KLH is an expensive protein that is not an economically viable alternative for developing this peptide as a commercial vaccine. Methods. The synthetic P0 peptide was chemically conjugated to the Bm86 protein obtained by fermentation of a recombinant Pichia pastoris yeast. This conjugate was characterized by SDS-PAGE analysis and mass spectrometry and was used in immunization and challenge experiments against different tick species. Results. SDS-PAGE analysis of this conjugate showed that it is very heterogeneous and that there are from 1 to 18 molecules of pP0 per molecule of Bm86. LC-MS/MS analysis and the combination of 4 different software showed that 49 of the 54 lysine residues and the N-terminal end of the Bm86 protein were partially conjugated to pP0. This conjugate pP0-Bm86 generated an immune response against the peptide when used as an antigen in immunization experiments of different animals. This immune response affected the viability and reproductive potential of different tick species. Conclusion, these results demonstrate that the P0 peptide is a broad spectrum antigen and permit its development as an anti-tick vaccine.
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