Non-hemagglutinating flaviviruses: molecular mechanisms for the emergence of new strains via adaptation to European ticks.

Tick-borne encephalitis virus (TBEV) causes human epidemics across Eurasia. Clinical manifestations range from inapparent infections and fevers to fatal encephalitis but the factors that determine disease severity are currently undefined. TBEV is characteristically a hemagglutinating (HA) virus; the ability to agglutinate erythrocytes tentatively reflects virion receptor/fusion activity. However, for the past few years many atypical HA-deficient strains have been isolated from patients and also from the natural European host tick, Ixodes persulcatus. By analysing the sequences of HA-deficient strains we have identified 3 unique amino acid substitutions (D67G, E122G or D277A) in the envelope protein, each of which increases the net charge and hydrophobicity of the virion surface. Therefore, we genetically engineered virus mutants each containing one of these 3 substitutions; they all exhibited HA-deficiency. Unexpectedly, each genetically modified non-HA virus demonstrated increased TBEV reproduction in feeding Ixodes ricinus, not the recognised tick host for these strains. Moreover, virus transmission efficiency between infected and uninfected ticks co-feeding on mice was also intensified by each substitution. Retrospectively, the mutation D67G was identified in viruses isolated from patients with encephalitis. We propose that the emergence of atypical Siberian HA-deficient TBEV strains in Europe is linked to their molecular adaptation to local ticks. This process appears to be driven by the selection of single mutations that change the virion surface thus enhancing receptor/fusion function essential for TBEV entry into the unfamiliar tick species. As the consequence of this adaptive mutagenesis, some of these mutations also appear to enhance the ability of TBEV to cross the human blood-brain barrier, a likely explanation for fatal encephalitis. Future research will reveal if these emerging Siberian TBEV strains continue to disperse westwards across Europe by adaptation to the indigenous tick species and if they are associated with severe forms of TBE.

Tick-borne encephalitis virus foci in Slovakia.

Tick-borne encephalitis (TBE) virus as a typical arbovirus relies on two types of hosts for its survival: ticks act both as virus vectors and reservoir hosts, and vertebrates amplify the virus infection by acting as a source of infection for feeding ticks. Longitudinal monitoring of TBE virus in ticks and vertebrate hosts including humans over a period of 40 years resulted in the identification of the areas of Slovakia where TBE virus is endemic. These are concentrated to the western, southern, and eastern parts of the country. Even with recently identified foci there is no evidence that the size and location of the natural TBE foci have changed significantly during the last decades. Numbers of diagnosed hospitalised cases of TBE in Slovakia vary from less than 20 to almost 100 cases annually with 54-89 cases in recent years. A part of these cases (33 cases during the last 5 years) are alimentary infections after drinking of raw goat and sheep milk.