In Europe, Ixodes ricinus is the most important vector of human infectious diseases, most notably Lyme borreliosis and tick-borne encephalitis virus. Multiple non-natural hosts of I. ricinus have shown to develop immunity after repeated tick bites. Tick immunity has also been shown to impair B. burgdorferi transmission. Most interestingly, multiple tick bites reduced the likelihood of contracting Lyme borreliosis in humans. A vaccine that mimics tick immunity could therefore potentially prevent Lyme borreliosis in humans. A yeast surface display library (YSD) of nymphal I. ricinus salivary gland genes expressed at 24, 48 and 72 h into tick feeding was constructed and probed with antibodies from humans repeatedly bitten by ticks, identifying twelve immunoreactive tick salivary gland proteins (TSGPs). From these, three proteins were selected for vaccination studies. An exploratory vaccination study in cattle showed an anti-tick effect when all three antigens were combined. However, immunization of rabbits did not provide equivalent levels of protection. Our results show that YSD is a powerful tool to identify immunodominant antigens in humans exposed to tick bites, yet vaccination with the three selected TSGPs did not provide protection in the present form. Future efforts will focus on exploring the biological functions of these proteins, consider alternative systems for recombinant protein generation and vaccination platforms and assess the potential of the other identified immunogenic TSGPs.

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

Center for Experimental and Molecular Medicine Amsterdam Multidisciplinary Lyme Borreliosis Center Amsterdam UMC Location Academic Medical Center University of Amsterdam Amsterdam The Netherlands

CIC bioGUNE Basque Research and Technology Alliance 48160 Derio Spain

Ikerbasque Basque Foundation for Science 48012 Bilbao Spain

Institute for Parasitology and Tropical Veterinary Medicine Freie Universität Berlin Berlin Germany

Section of Infectious Diseases Department of Internal Medicine Yale University New Haven CT USA

UCD Conway Institute University College Dublin Belfield Dublin 4 Ireland

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