A chemical conjugate of the tick P0 peptide is efficacious against<i>Amblyomma mixtum</i>

Rodríguez-Mallón, Alina; Guzmán, Pedro E. Encinosa; Soto, Yamil Bello; Perdomo, Karen Rosales; Espinosa, Carlos Montero; Vargas, Marlene; Estrada, Mario Pablo

doi:10.1111/tbed.13455

Cited by 14 publications

(19 citation statements)

References 14 publications

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“…More recently, an antigen based on the peptide of the P0 acidic ribosomal protein of ticks from amino acid 282 to 301 (pP0) chemically conjugated to keyhole limpet hemocyanin (KLH) of Megathura crenulata was evaluated against Rhipicephalus spp. Ticks, showing around 90% of efficacy [ 10 , 11 ], and also against Amblyomma mixtum [ 12 ] and Ixodes ricinus ticks (manuscript in preparation). However, the pathway from concept proof in controlled experiments to the development of a commercial vaccine is a long, difficult and costly process.…”

Section: Introductionmentioning

confidence: 99%

Functional and Mass Spectrometric Evaluation of an Anti-Tick Antigen Based on the P0 Peptide Conjugated to Bm86 Protein

et al. 2020

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A synthetic 20 amino acid peptide of the ribosomal protein P0 from ticks, when conjugated to keyhole limpet hemocyanin from Megathura crenulata and used as an immunogen against Rhipicephalus microplus and Rhipicephalus sanguineus s.l. species, has shown efficacies of around 90%. There is also experimental evidence of a high efficacy of this conjugate against Amblyomma mixtum and Ixodes ricinus species, which suggest that this antigen could be a good broad-spectrum anti-tick vaccine candidate. In this study, the P0 peptide (pP0) was chemically conjugated to Bm86 as a carrier protein. SDS-PAGE analysis of this conjugate demonstrated that it is highly heterogeneous in size, carrying from 1 to 18 molecules of pP0 per molecule of Bm86. Forty-nine out of the 54 lysine residues and the N-terminal end of Bm86 were found partially linked to pP0 by using LC-MS/MS analysis and the combination of four different softwares. Several post-translational modifications of Bm86 protein were also identified by mass spectrometry. High immunogenicity and efficacy were achieved when dogs and cattle were vaccinated with the pP0–Bm86 conjugate and challenged with R. sanguineus s.l. and R. microplus, respectively. These results encourage the development of this antigen with promising possibilities as an anti-tick vaccine.

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Section: Introductionmentioning

confidence: 99%

Functional and Mass Spectrometric Evaluation of an Anti-Tick Antigen Based on the P0 Peptide Conjugated to Bm86 Protein

et al. 2020

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“…In our experience, the pP0 alone or expressed as a fusion protein was not able to induce a strong host immune response (data not shown). The best immunological response and efficacy results obtained with pP0-based vaccines have been obtained after chemical conjugation of this peptide to highly aggregated carrier proteins with very well-ordered quaternary structure and very high molecular masses such as KLH [ 9 – 11 ] and the particulate Bm86 protein expressed intracellularly by Pichia pastoris yeast [ 15 , 61 ]. There are many examples in literature highlighting the role of multimerization and/or aggregation in the generation of an immune response (65, 66), and for the vaccines’ development, the immune response enhancement by inducing antigen aggregation in different ways has an important implication.…”

Section: Resultsmentioning

confidence: 99%

“…Chemical conjugation of a 20-amino acid peptide from P0 acidic ribosomal protein of ticks (pP0) to keyhole limpet hemocyanin (KLH) from Megathura crenulata has shown to be effective against different tick species when used for host vaccination [ 9 – 11 ].…”

Section: Introductionmentioning

confidence: 99%

Synthesis, LC-MS/MS analysis, and biological evaluation of two vaccine candidates against ticks based on the antigenic P0 peptide from R. sanguineus linked to the p64K carrier protein from Neisseria meningitidis

et al. 2021

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A peptide from the P0 acidic ribosomal protein (pP0) of ticks conjugated to keyhole limpet hemocyanin from Megathura crenulata has shown to be effective against different tick species when used in host vaccination. Turning this peptide into a commercial anti-tick vaccine will depend on finding the appropriate, technically and economically feasible way to present it to the host immune system. Two conjugates (p64K-Cys 1 pP0 and p64K-βAla 1 pP0) were synthesized using the p64K carrier protein from Neisseria meningitidis produced in Escherichia coli , the same cross-linking reagent, and two analogues of pP0. The SDS-PAGE analysis of p64K-Cys 1 pP0 showed a heterogeneous conjugate compared to p64K-βAla 1 pP0 that was detected as a protein band at 91kDa. The pP0/p64K ratio determined by MALDI-MS for p64K-Cys 1 pP0 ranged from 1 to 8, being 3-5 the predominant ratio, while in the case of p64K-βAla 1 pP0 this ratio was 5-7. Cys 1 pP0 was partially linked to 35 out of 39 Lys residues and the N-terminal end, while βAla 1 pP0 was mostly linked to the six free cysteine residues, to the N-terminal end, and, in a lesser extent, to Lys residues. The assignment of the conjugation sites and side reactions were based on the identification of type 2 peptides. Rabbit immunizations showed the best anti-pP0 titers and the highest efficacy against Rhipicephalus sanguineus ticks when the p64K-Cys 1 pP0 was used as vaccine antigen. The presence of high molecular mass aggregates observed in the SDS-PAGE analysis of p64K-Cys 1 pP0 could be responsible for a better immune response against pP0 and consequently for its better efficacy as an anti-tick vaccine. Graphical abstract

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“…The BM86 antigen, which is the base of the commercial vaccines against R. microplus , was ineffective against nymphs and adults of A. cajennense sensu lato when calves were used as hosts ( 68 ). In an experiment conducted with larvae on rabbits, the tick P0 peptide showed 54% efficacy against Amblyomma mixtum , which is another species of the A. cajennense complex ( 69 ).…”

Section: Discussionmentioning

confidence: 99%

Amblyomma sculptum Salivary Protease Inhibitors as Potential Anti-Tick Vaccines

et al. 2021

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Amblyomma sculptum is the main tick associated with human bites in Brazil and the main vector of Rickettsia rickettsii, the causative agent of the most severe form of Brazilian spotted fever. Molecules produced in the salivary glands are directly related to feeding success and vector competence. In the present study, we identified sequences of A. sculptum salivary proteins that may be involved in hematophagy and selected three proteins that underwent functional characterization and evaluation as vaccine antigens. Among the three proteins selected, one contained a Kunitz_bovine pancreatic trypsin inhibitor domain (named AsKunitz) and the other two belonged to the 8.9 kDa and basic tail families of tick salivary proteins (named As8.9kDa and AsBasicTail). Expression of the messenger RNA (mRNA) encoding all three proteins was detected in the larvae, nymphs, and females at basal levels in unfed ticks and the expression levels increased after the start of feeding. Recombinant proteins rAs8.9kDa and rAsBasicTail inhibited the enzymatic activity of factor Xa, thrombin, and trypsin, whereas rAsKunitz inhibited only thrombin activity. All three recombinant proteins inhibited the hemolysis of both the classical and alternative pathways; this is the first description of tick members of the Kunitz and 8.9kDa families being inhibitors of the classical complement pathway. Mice immunization with recombinant proteins caused efficacies against A. sculptum females from 59.4% with rAsBasicTail immunization to more than 85% by immunization with rAsKunitz and rAs8.9kDa. The mortality of nymphs fed on immunized mice reached 70–100%. Therefore, all three proteins are potential antigens with the possibility of becoming a new tool in the control of A. sculptum.

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A chemical conjugate of the tick P0 peptide is efficacious againstAmblyomma mixtum

Cited by 14 publications

References 14 publications

Functional and Mass Spectrometric Evaluation of an Anti-Tick Antigen Based on the P0 Peptide Conjugated to Bm86 Protein

Functional and Mass Spectrometric Evaluation of an Anti-Tick Antigen Based on the P0 Peptide Conjugated to Bm86 Protein

Synthesis, LC-MS/MS analysis, and biological evaluation of two vaccine candidates against ticks based on the antigenic P0 peptide from R. sanguineus linked to the p64K carrier protein from Neisseria meningitidis

Amblyomma sculptum Salivary Protease Inhibitors as Potential Anti-Tick Vaccines

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