Few studies have examined tick proteomes, how they adapt to their environment, and their roles in the parasite-host interactions that drive tick infestation and pathogen transmission. Here we used a proteomics approach to screen for biologically and immunologically relevant proteins acting at the tick-host interface during tick feeding and, as proof of principle, measured host antibody responses to some of the discovered candidates. We used a label-free quantitative proteomic workflow to study salivary proteomes of (i) wild Ixodes ricinus ticks fed on different hosts, (ii) wild or laboratory ticks fed on the same host, and (iii) adult ticks cofed with nymphs. Our results reveal high and stable expression of several protease inhibitors and other tick-specific proteins under different feeding conditions. Most pathways functionally enriched in sialoproteomes were related to proteolysis, endopeptidase, and amine-binding activities. The generated catalogue of tick salivary proteins enabled the selection of six candidate secreted immunogenic peptides for rabbit immunizations, three of which induced strong and durable antigen-specific antibody responses in rabbits. Furthermore, rabbits exposed to ticks mounted immune responses against the candidate peptides/proteins, confirming their expression at the tick-vertebrate interface. Our approach provides insights into tick adaptation strategies to different feeding conditions and promising candidates for developing antitick vaccines or markers of exposure of vertebrate hosts to tick bites.

Departamento de Genética Facultad de Ciencias Universidad de Granada Granada Spain

Faculty of Science University of South Bohemia Ceske Budejovice Czech Republic

Institute for Immunology University Medical Center of the Johannes Gutenberg University Mainz Mainz Germany

Institute of Medical Biostatistics Epidemiology and Informatics University Medical Center of the Johannes Gutenberg University Mainz Mainz Germany

Institute of Parasitology Biology Centre Czech Academy of Sciences Ceske Budejovice Czechia

Laboratory de Bioinformática Centro de Investigación Biomédica PTS Instituto de Biotecnología Granada Spain

University of Antwerp Wilrijk Belgium

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The immune factors involved in the rapid clearance of bacteria from the midgut of the tick Ixodes ricinus

An Unusual Two-Domain Thyropin from Tick Saliva: NMR Solution Structure and Highly Selective Inhibition of Cysteine Cathepsins Modulated by Glycosaminoglycans

Insight Into the Dynamics of the Ixodes ricinus Nymphal Midgut Proteome