Vaccination against tick-borne encephalitis (TBE) is based on the use of formalin-inactivated, culture-derived whole-virus vaccines. Immune response following vaccination is primarily directed to the viral envelope (E) protein, the major viral surface antigen. In Europe, two TBE vaccines are available in adult and pediatric formulations, namely FSME-IMMUN® (Pfizer) and Encepur® (GlaxoSmithKline). Herein, we analyzed the content of these vaccines using mass spectrometry (MS). The MS analysis revealed that the Encepur vaccine contains not only proteins of the whole virus particle, but also viral non-structural protein 1 (NS1). MS analysis of the FSME-IMMUN vaccine failed due to the high content of human serum albumin used as a stabilizer in the vaccine. However, the presence of NS1 in FSME-IMMUN was confirmed by immunization of mice with six doses of this vaccine, which led to a robust anti-NS1 antibody response. NS1-specific Western blot analysis also detected anti-NS1 antibodies in sera of humans who received multiple doses of either of these two vaccines; however, most vaccinees who received ≤3 doses were negative for NS1-specific antibodies. The contribution of NS1-specific antibodies to protection against TBE was demonstrated by immunization of mice with purified NS1 antigen, which led to a significant (p < 0.01) prolongation of the mean survival time after lethal virus challenge. This indicates that stimulation of anti-NS1 immunity by the TBE vaccines may increase their protective effect.

Central European Institute of Technology Masaryk University Kamenice 753 5 CZ 62500 Brno Czech Republic

Faculty of Science Masaryk University Kamenice 753 5 CZ 62500 Brno Czech Republic

Faculty of Science University of South Bohemia Branisovska 31 CZ 37006 Ceske Budejovice Czech Republic

Hospital Ceske Budejovice B Nemcove 585 54 370 01 Ceske Budejovice Czech Republic

Institute of Parasitology Biology Centre of the Czech Academy of Sciences Branisovska 31 CZ 37006 Ceske Budejovice Czech Republic

Royal Liverpool University Hospital Prescot St Liverpool L7 8XP UK

The Jack Copland Centre Scottish National Blood Transfusion Service 52 Research Avenue North Edinburgh EH14 4BE UK

Veterinary Research Institute Hudcova 70 CZ 62100 Brno Czech Republic

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Dynamics of Whole Virus and Non-Structural Protein 1 (NS1) IgG Response in Mice Immunized with Two Commercial Tick-Borne Encephalitis Vaccines

History of Arbovirus Research in the Czech Republic