Tick-borne encephalitis virus (TBEV) is a flavivirus that causes human neuroinfections and represents a growing health problem. The human monoclonal antibody T025 targets envelope protein domain III (EDIII) of TBEV and related tick-borne flaviviruses, potently neutralizing TBEV in vitro and in preclinical models, representing a promising candidate for clinical development. We demonstrate that TBEV escape in the presence of T025 or T028 (another EDIII-targeting human monoclonal antibody) results in virus variants of reduced pathogenicity, characterized by distinct sets of amino acid changes in EDII and EDIII that are jointly needed to confer resistance. EDIII substitution K311N impairs formation of a salt bridge critical for T025-epitope interaction. EDII substitution E230K is not on the T025 epitope but likely induces quaternary rearrangements of the virus surface because of repulsion of positively charged residues on the adjacent EDI. A combination of T025 and T028 prevents virus escape and improves neutralization.

California Institute of Technology Pasadena CA USA

Institute for Research in Biomedicine Università della Svizzera Italiana Bellinzona Switzerland

Institute for Research in Biomedicine Università della Svizzera Italiana Bellinzona Switzerland; Swiss Institute of Bioinformatics Lausanne Switzerland

Laboratory of Molecular Immunology The Rockefeller University New York NY USA

Laboratory of Molecular Immunology The Rockefeller University New York NY USA; Howard Hughes Medical Institute New York NY USA

Veterinary Research Institute Brno Czech Republic; Department of Experimental Biology Faculty of Science Masaryk University Brno Czech Republic

Veterinary Research Institute Brno Czech Republic; Department of Experimental Biology Faculty of Science Masaryk University Brno Czech Republic; Faculty of Science University of South Bohemia Ceske Budejovice Czech Republic

Veterinary Research Institute Brno Czech Republic; Institute of Parasitology Biology Centre of the Czech Academy of Sciences Ceske Budejovice Czech Republic

Veterinary Research Institute Brno Czech Republic; Institute of Parasitology Biology Centre of the Czech Academy of Sciences Ceske Budejovice Czech Republic; Department of Experimental Biology Faculty of Science Masaryk University Brno Czech Republic; Department of Pharmacology and Pharmacy Faculty of Veterinary Medicine University of Veterinary Sciences Brno Czech Republic

Veterinary Research Institute Brno Czech Republic; Institute of Parasitology Biology Centre of the Czech Academy of Sciences Ceske Budejovice Czech Republic; Department of Experimental Biology Faculty of Science Masaryk University Brno Czech Republic; Joint Faculty of Veterinary Medicine Yamaguchi University Yamaguchi City Japan

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