Tick-borne Encephalitis Virus (TBEV) 11084 Tick-borne Encephalitis Tick-borne encephalitis virus (TBEV) is a virus of the family Flaviviridae and has three subtypes: European or Western tick-borne encephalitis virus, Siberian tick-borne encephalitis virus and Far eastern tick-borne encephalitis virus (formerly known as Russian Spring Summer encephalitis virus). TBEV causes tick-borne encephalitis, a human viral disease of the central nervous system, typically presenting in meningitis, encephalitis or meningoencephilitis. Ixodid Ticks are the vector and reservoir for TBEV, with the main hosts being rodents, with humans being accidental hosts. The disease is endemic to Europe, Russia and Asia. Baboon Papio cynocephalus 9556 Bank vole Clethrionomys glareolus 447135 Bear Ursus americanus 9643 Birds Passeroidea 175121 Brown Trout Salmo trutta 8032 Buffalo Bison bison 9901 Carnivores Vulpes 9625 Cat Felis catus 9685 Catfishes Siluriformes 7995 Cattle Bos taurus 9913 Chicken Gallus gallus 9031 Chimpanzee Pan troglodytes 9598 chinchillas Chinchillidae 10150 Copper Pheasant Syrmaticus soemmerringii 9067 Deer Cervus elaphus 9860 Deer mouse Peromyscus maniculatus 10042 Dog Canis familiaris 9615 Ducks Anas 8835 Ferret Mustela putorius furo 9669 Fish Hyperotreti 117565 Gerbil Gerbillina 10045 Goat Capra hircus 9925 Gray wolf Canis lupus 9612 Guinea pig Cavia porcellus 10141 Hamster Mesocricetus auratus 10036 Horse Equus caballus 9796 Human Homo sapiens 9606 Macaque Macaca fascicularis 9541 Mongolian Gerbil Meriones unguiculatus 10047 Monkey Platyrrhini 9479 Mouse Mus musculus 10090 None None Parrot Psittacidae 9224 Pig Sus scrofa 9823 Rabbit Oryctolagus cuniculus 9986 Rainbow trout Oncorhynchus mykiss 8022 Rat Rattus 10114 Raven Corvus corax 56781 sei whale Balaenoptera borealis 9768 Sheep Ovis aries 9940 Squirrel Spermophilus richardsonii 37591 Tree shrew Tupaiidae 9393 Trouts, salmons & chars Salmoninae 504568 Turkey Meleagris gallopavo 9103 Vole Microtus ochrogaster 79684 Water buffalo Bubalus bubalis 391902 Encepur GlaxoSmithKline Pfizer Inactivated or "killed" vaccine Licensed Intramuscular injection (i.m.) Encepur is a -virus formalin-inactivated TBEV vaccines avaliable in the European Union and European Economic Area. Vaccination induces neutralizing antibodies directed against the main antigenic determinant of the viral particle, the E protein.(Salat et al., 2020) ncepur Adults vaccines were processed using filter-aided sample preparation (FASP). , protein cysteine disulfide bonds were reduced in 0.1M dithiothreitol (DTT) in 8 M urea/0.1 M Tris/HCl (pH 8.5), and 50 mM iodoacetamide was used to alkylate cysteine residues (Salat et al., 2020). Intramuscular injection (i.m.) Proteins of whole virus particle and viral non structural protein 1 (NS1) and E protein.(Salat et al., 2020) Recombinant protein preparation Inactivated Recombinant protein preparation Inactivated FSME-IMMUN Tick-borne encephalitis vaccine, inactivated FSME-IMMUN Baxter VO_0010714 Inactivated or "killed" vaccine Licensed Intramuscular injection (i.m.) Canada Aluminum hydroxide FSME-IMMUN should be stored between +2°C to +8°C. Intramuscular injection (i.m.) licensed Tick-borne encephalitis human vaccine Generic Unknown VO_0012190 Inactivated or "killed" vaccine Licensed A generic representation of vaccines used to prevent Tick-borne Encephalitis in humans, most commonly formulated as inactivated (killed) whole-virus vaccines. These vaccines are administered to induce immunity without the risk of causing disease. TBEV DNA vaccine encoding E protein VO_0004154 DNA vaccine Research pCMV-β [Ref1249:Aberle et al., 1999] Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction BALB/c Groups of 16 BALB/c mice each received primary and booster immunizations spaced 4 wk apart. Plasmids were purified and administered i.m. (100 µg/dose) or by particle bombardment with DNA-coated gold beads (2 µg/dose) using the helium-powered Helios Gene Gun delivery system. Intramuscular inoculation of DNA in saline was done as a 100-µl injection in the right quadriceps. For Gene Gun inoculation, 1.0 µg of the same DNA was coupled to 0.5 mg of 1.0-µm-diameter gold particles, as recommended by the manufacturer (Bio-Rad). DNA-coated microcarriers were delivered into the abdominal epidermis of mice using the Gene Gun at a helium pressure setting of 400 psi. Mice immunized with the commercial TBE vaccine received 0.2 ml s.c. inoculations containing 1 µg formalin-inactivated virus (FSME Immun Inject, Baxter-Immuno, Vienna, Austria) (Aberle et al., 1999). The plasmid construct encoding a secreted subviral particle, which carries multiple copies of the protective Ag on its surface, was superior to the other constructs in terms of extent and functionality of the Ab response as well as protection against virus challenge (Aberle et al., 1999). Mice were subsequently challenged by i.p. inoculation with 1000 LD50 of the highly mouse-pathogenic TBE virus strain Hypr. (Aberle et al., 1999). TBEV Virus Like Particle Vaccine Virus Particle Vaccine Research Intramuscular injection (i.m.) Virus-like particle-based vaccine co-expressing the structural (Core/prM/E) and non-structural (NS2B/NS3Pro) proteins of TBEV confers complete protection against TBEV in mice. (Tang et al., 2023) Expi293 cells were diluted to a density of 1.5 × 106 cells/mL before transfection with pcDNA3.1-NS2B/NS3Pro and pcDNA3.1-C/prM/E at a ratio of 1:2, then the cells were cultivated at 37°C in an atmosphere of 5% CO2, with agitation at 150 rpm. For the production of VLPs, the plasmids (480 μg) were added to polyethylenimine hydrochloride (1,350 μg; Polysciences) in 10 mL of Expi293 medium and incubated for 10 min, the mixture was then added to 470 mL of cell solution. The transfected cells were harvested at 4 days post-transfection and the supernatant was collected by two successive centrifugation steps at 400 ×g for 10 min at 4°C, then 10,000 ×g for 10 min at 4°C. The proteins in the supernatant were precipitated using 8% (wt/vol) polyethylene glycol 8000 and incubated overnight at 4°C. (Tang et al., 2023) Intramuscular injection (i.m.) Atructural (Core/prM/E) and non-structural (NS2B/NS3Pro) proteins of TBEV (Tang et al., 2023) The results of ELISA demonstrated that vaccination with the VLPs stimulated a high total serum IgG response, and the prime antibody response increased by 2.5-fold following boost immunization (Fig. 2C). Additionally, the antibodies elicited by the VLPs significantly neutralized both TBEV-FE and TBEV-Eu infections in BHK-21 cells, with inhibition rates reaching up to 90%. Altogether, these findings suggested that the VLPs exhibited superior immunogenicity and induced high levels of humoral immunity (IgG and NAb) against different subtypes of TBEV. (Tang et al., 2023) Eight-week-old male C57BL/6 mice were used for the immunization and viral neutralization experiments. The mice (n=5 per group) were immunized by twice intramuscular injections with PBS or 10 μg of VLPs.Six to eight-week-old male IFNAR-/- mice were used for immunization and challenge experiments. The mice (n=10 per group) were immunized with PBS or 10 μg of VLPs. (Tang et al., 2023) The infected mice displayed some abnormal behaviors, such as listlessness, circling and freezing, and all the infected mice died within 8 dpi. In contrast, immunized mice survived the challenge, and their weights did not decrease significantly. The viremia of the control group was notably high (7.9×104 TCID50/mL), with average values in the brain and intestinal tissues reaching 1.0×10^7 and 5.0×10^5 TCID50/mL, respectively. In contrast, the mice that had been immunized with the VLPs did not exhibit any neurological symptoms and the viral loads in the serum samples, brain, and intestinal tissues were negligible or undetectable. (Tang et al., 2023) VLP-immunized IFNAR−/− mice were challenged with a lethal dose (3.2×106 PFU) of the WH2012 strain of TBEV by intramuscular injection after two weeks post-boost immunization with 10 μg VLPs. (Tang et al., 2023) Tick-borne Encephalitis Virus C protein mutant vaccine VO_0004278 Live, attenuated vaccine Research subcutaneous injection subcutaneous injection Gene mutation This C protein mutant is from Tick-borne encephalitis virus (Kofler et al., 2002). A C protein mutant is attenuated in mice (Kofler et al., 2002). A C protein mutant induced protection in mice from challenge with wild type TBEV (Kofler et al., 2002). C protein Tick-borne encephalitis virus 206570298 CDD:110034 11084 ? capsid protein 35 Flavivirus capsid protein C; pfam01003 >gi|206570298|gb|ACI12930.1| capsid protein [Tick-borne encephalitis virus] MARKAILKGKGGGPPRRVSKETAKKTRQSRVQMPN Virmugen A virus with a mutation in protein C is attenuated in mice and induced protection from challenge with wild type TBE virus [Ref1783:Kofler et al., 2002]. E Tick-borne Encephalitis Virus (TBEV) CAC94472 CDD:279241 CDD:213392 CDD:213897 GOA:Q8UYS5 HSSP:1SVB InterPro:IPR000336 InterPro:IPR011998 InterPro:IPR011999 InterPro:IPR013755 InterPro:IPR013756 InterPro:IPR014756 UniProtKB/TrEMBL:Q8UYS5 11084 E protein 7.56 42769 494 Flavivirus glycoprotein, central and dimerization domains; pfam00869 >CAC94472.1 E protein, partial [Tick-borne encephalitis virus] TQGTTRVTLVLELGGCVTITAEGKPSMDVWLDSIYQENPAKTREYCLHAKLSDTKVAARCPTMGPAVLAE EHQSGTVCKRDQSDRGWGNHCGLFGKGSIVTCVKAACEAKKKATGHVYDANKIVYTVKVEPHTGDYVAAN ETHSGRKTASFTVSSEKTILNMGDYGDVSLLCRVASGVDLAQTVILELDKTLEHLPTAWQVHRDWFNDLA LPWKHEGAQHWNNAERLVEFGAPHAVKMDVYNLGDQTGVLLKSLAGVPVAHIDGTKYHLKSGHVTCEVGL EKLKMKGLTYTMCDKTKFTWKRTPTDSGHDTVVMEVTFSGTKPCRIPVRAVAHGSPDVNVAMLITPNPTI ENNGGGFIEMQLPPGDNIIYVGELSHQWFQKGSSIGRVFQKTRKGIERLTVIGEHAWDFGSTGGFLTSVG KAL Protective antigen [Ref4970:Stenkova et al., 2017] NS1 Tick-borne Encephalitis Virus (TBEV) CAA44678 CDD:213897 CDD:279316 InterPro:IPR001157 UniProtKB/TrEMBL:Q88482 11084 unnamed 6.07 38586.51 441 flavivirus envelope glycoprotein E, stem/anchor domain; TIGR04240 >CAA44678.1 NS1, partial [Tick-borne encephalitis virus] RNPTMSMSFLLAGGLVLAMTLGVGADVGCAVDTERMELRCGEGLVVWREVSEWYDNYAYYPETPGALASA IKETFEEGSCGVVPQNRLEMAMWRSSVTVLNLALAEGEANLTVVVDKFDPTDYRGGVSGLLRKGKDIKAS WKSWGHSIIWSTPEAPRRFMVGTEGQSECPLERRKTGVFTVAEFGVGLRTKVFLDFRQEPTHECDTGVMG AAVKNGMAIHTDQSLWMRSMKNDTGTYIVELLVTDLRNCSWPASHTIDNADVVDSELLLPASLAGPRSWY NRIPGYSEQVKGPWKYTPIRVIREECPGTTVTINAKCDKRGASVRSTTESGKVIPEWCCRACTMPPVTFR TGADCWYAMEIRPVHDQGGLVRSMVVA Protective antigen [Ref5011:Dmitriev et al., 1996] TBEVgp1 polyprotein Tick-borne encephalitis virus VO_0011245 1489719 9628432 TBEVgp1 U27495 NP_043135 11084 132 10376 + polyprotein 8.16 347270.05 3414 >NC_001672.1:132-10376 Tick-borne encephalitis virus, complete genome GATGGTCAAGAAGGCCATCCTGAAAGGTAAGGGGGGCGGTCCCCCTCGACGAGTGTCGAAAGAGACCGCA ACGAAGACGCGTCAACCCAGAGTCCAAATGCCAAATGGGCTTGTGTTGATGCGCATGATGGGGATCTTGT GGCATGCCGTAGCCGGCACCGCGAGAAACCCCGTATTGAAGGCGTTCTGGAACTCAGTCCCTCTGAAACA GGCAACAGCAGCACTGCGGAAGATCAAAAGGACGGTGAGTGCCCTAATGGTTGGCTTGCAAAAACGTGGG AAAAGGAGGTCAGCGACGGACTGGATGAGCTGGTTGCTGGTCATCACTCTGTTGGGGATGACGCTTGCTG CAACGGTGAGGAAAGAAAGGGATGGCTCAACTGTGATCAGAGCTGAAGGAAAGGATGCAGCAACTCAGGT GCGTGTGGAGAATGGCACCTGTGTGATCCTGGCTACTGACATGGGGTCATGGTGTGATGATTCACTGTCC TATGAGTGTGTGACCATAGATCAAGGAGAAGAGCCTGTTGACGTGGATTGTTTTTGCCGGAACGTTGATG GAGTCTATCTGGAGTACGGACGCTGTGGGAAACAGGAAGGCTCACGGACAAGGCGCTCAGTGCTGATCCC ATCCCATGCTCAGGGAGAGCTGACGGGAAGGGGACACAAATGGCTAGAAGGAGACTCGCTGCGAACACAC CTTACTAGAGTTGAGGGATGGGTCTGGAAGAACAAGCTACTTGCCTTGGCAATGGTTACCGTTGTGTGGT TGACCCTGGAGAGTGTGGTGACCAGGGTTGCCGTTCTTGTTGTGCTCCTGTGTTTGGCACCGGTTTACGC TTCGCGTTGCACACACTTGGAAAACAGGGACTTTGTGACTGGTACTCAGGGGACTACGAGGGTCACCTTG GTGCTGGAACTGGGTGGATGTGTTACTATAACAGCTGAGGGGAAGCCTTCAATGGATGTGTGGCTTGACG CCATTTACCAGGAGAACCCTGCTAAGACACGTGAGTACTGTTTACACGCCAAGTTGTCGGACACTAAGGT TGCAGCCAGATGCCCAACAATGGGACCAGCCACTTTGGCTGAAGAACACCAGGGTGGCACAGTGTGTAAG AGAGATCAGAGTGATCGAGGCTGGGGCAACCACTGTGGACTGTTTGGAAAGGGTAGCATTGTGGCCTGTG TCAAGGCGGCTTGTGAGGCAAAAAAGAAAGCCACAGGACATGTGTACGACGCCAACAAAATAGTGTACAC GGTCAAAGTCGAACCACACACGGGAGACTATGTTGCCGCAAACGAGACACATAGTGGGAGGAAGACGGCA TCCTTCACAATTTCTTCAGAGAAAACCATTTTGACTATGGGTGAGTATGGAGATGTGTCTTTGTTGTGCA GGGTCGCTAGTGGCGTTGACTTGGCCCAGACCGTCATCCTTGAGCTTGACAAGACAGTGGAACACCTTCC AACGGCTTGGCAGGTCCATAGGGACTGGTTCAATGATCTGGCTCTGCCATGGAAACATGAGGGAGCGCAA AACTGGAACAACGCAGAAAGACTGGTTGAATTTGGGGCTCCTCACGCTGTCAAGATGGACGTGTACAACC TCGGAGACCAGACTGGAGTGTTACTGAAGGCTCTCGCTGGGGTTCCTGTGGCACACATTGAGGGAACCAA GTACCACCTGAAGAGTGGCCACGTGACCTGCGAAGTGGGACTGGAAAAACTGAAGATGAAAGGTCTTACG TACACAATGTGTGACAAAACAAAGTTCACATGGAAGAGAGCTCCAACAGACAGTGGGCATGATACAGTGG TCATGGAAGTCACATTCTCTGGAACAAAGCCCTGTAGGATCCCAGTCAGGGCAGTGGCACATGGATCTCC AGATGTGAACGTGGCCATGCTGATAACGCCAAACCCAACAATTGAAAACAATGGAGGTGGCTTCATAGAG ATGCAGCTGCCCCCAGGGGATAACATCATCTATGTTGGGGAACTGAGTCATCAATGGTTCCAAAAAGGGA GCAGCATCGGAAGGGTTTTCCAAAAGACCAAGAAAGGCATAGAAAGACTGACAGTGATAGGAGAGCACGC CTGGGACTTCGGTTCTGCTGGAGGCTTTCTGAGTTCAATTGGGAAGGCGGTACATACGGTCCTTGGTGGC GCTTTCAACAGCATCTTCGGGGGAGTGGGGTTTCTACCAAAACTTTTATTAGGAGTGGCATTGGCTTGGT TGGGCCTGAACATGAGAAACCCTACAATGTCCATGAGCTTTCTCTTGGCTGGAGGTCTGGTCTTGGCCAT GACCCTTGGAGTGGGGGCGGATGTTGGTTGCGCTGTGGACACGGAACGAATGGAGCTCCGCTGTGGCGAG GGCCTGGTCGTGTGGAGAGAGGTCTCAGAATGGTATGACAATTATGCCTACTACCCGGAGACACCGGGGG CCCTTGCATCAGCCATAAAGGAGACATTTGAGGAGGGAAGCTGTGGTGTAGTCCCCCAGAACAGGCTCGA GATGGCCATGTGGAGAAGCTCGGTCACAGAGCTGAATTTGGCTCTGGCGGAAGGGGAGGCAAATCTCACA GTGGTGGTGGACAAGTTTGACCCCACTGACTACCGAGGTGGTGTCCCTGGTTTACTGAAAAAAGGAAAGG ACATAAAAGTCTCCTGGAAAAGCTGGGGCCATTCAATGATCTGGAGCATTCCTGAGGCCCCCCGTCGCTT CATGGTGGGCACGGAAGGACAAAGTGAGTGTCCCCTAGAGAGACGGAAGACAGGTGTTTTCACGGTGGCA GAATTCGGGGTTGGCCTGAGAACAAAGGTCTTCTTGGATTTCAGACAGGAACCAACACATGAGTGTGACA CAGGAGTGATGGGAGCTGCAGTCAAGAACGGCATGGCAATCCACACAGATCAAAGTCTCTGGATGAGATC AATGAAAAATGACACAGGCACTTACATAGTTGAACTTTTGGTCACTGACCTGAGGAACTGCTCATGGCCT GCTAGCCACACTATCGATAATGCTGACGTGGTGGACTCAGAGTTATTCCTTCCGGCGAGCCTGGCAGGAC CCAGATCCTGGTACAACAGGATACCTGGCTATTCAGAACAGGTGAAAGGGCCATGGAAGTACACGCCTAT CCGAGTTATCAGAGAGGAGTGTCCCGGCACGACCGTTACCATCAACGCCAAGTGTGACAAAAGAGGAGCA TCTGTGAGGAGTACCACAGAGAGTGGCAAGGTTATCCCAGAATGGTGCTGCCGAGCGTGCACAATGCCAC CAGTGACGTTCCGGACTGGAACTGATTGCTGGTATGCCATGGAAATACGGCCAGTCCATGACCAGGGGGG GCTTGTTCGCTCAATGGTGGTTGCGGACAACGGTGAATTACTTAGTGAGGGAGGAGTCCCCGGAATAGTG GCATTGTTTGTGGTCCTTGAATACATCATCCGTAGGAGGCCCTCCACGGGAACGACGGTTGTGTGGGGGG GTATCGTCGTTCTCGCTCTGCTTGTCACCGGGATGGTCAGGATAGAGAGCCTGGTGCGCTATGTGGTGGC AGTGGGGATCACATTCCACCTTGAGCTGGGGCCAGAGATCGTGGCCTTGATGCTACTCCAGGCTGTGTTT GAGCTGAGGGTGGGTTTGCTCAGCGCATTTGCATTGCGCAGAAGCCTCACCGTCCGAGAGATGGTGACCA CCTACTTTCTTTTGCTGGTCCTGGAATTGGGGCTGCCGGGTGCGAGCCTTGAGGAGTTCTGGAAATGGGG TGATGCACTGGCCATGGGGGCGCTGATATTCAGGGCTTGCACGGCAGAAGGAAAGACTGGAGCGGGGCTT TTGCTCATGGCTCTCATGACACAGCAGGATGTGGTGACTGTGCACCATGGACTGGTGTGCTTCCTAAGTG TAGCTTCGGCATGCTCGGTCTGGAGGCTGCTCAAGGGACACAGAGAGCAGAAGGGATTGACCTGGGTTGT CCCCCTGGCTGGGTTGCTTGGGGGAGAGGGCTCTGGAATCAGACTGCTGGCGTTTTGGGAGTTGTCAGCG CACAGAGGAAGACGATCTTTCAGTGAACCACTAACTGTGGTAGGAGTCATGCTAACACTGGCCAGCGGCA TGATGCGACACACTTCCCAGGAGGCTCTCTGTGCACTCGCAGTGGCCTCGTTTCTCTTGCTGATGCTGGT GCTGGGGACAAGAAAGATGCAGCTGGTTGCCGAATGGAGTGGCTGCGTTGAATGGTATCCGGAACTAGTG AATGAGGGTGGAGAGGTTAGCCTGCGGGTCCGGCAGGACGCGATGGGTAACTTTCACTTGACTGAGCTCG AGAAAGAAGAGAGAATGATGGCTTTTTGGCTGATTGCCGGCTTGGCAGCTTCGGCCATTCACTGGTCAGG CATTCTTGGTGTGATGGGACTGTGGACGCTCACGGAAATGCTGAGGTCATCCCGAAGGTCTGACCTGGTT TTCTCTGGACAGGGGGGTCGAGAGCGTGGTGACAGACCTTTCGAGGTTAAGGACGGTGTCTACAGGATTT TTAGCCCCGGCTTGTTCTGGGGTCAGAACCAGGTGGGAGTTGGCTACGGTTCCAAGGGTGTCTTGCACAC GATGTGGCACGTGACGAGAGGAGCGGCGCTGTCTATTGATGACGCTGTGGCCGGTCCCTACTGGGCTGAT GTGAGGGAAGATGTCGTGTGTTACGGAGGAGCCTGGAGCCTGGAGGAAAAATGGAAAGGTGAAACAGTAC AGGTCCATGCCTTCCCACCGGGGAGGGCGCATGAGGTGCATCAGTGCCAGCCTGGGGAGTTGATCCTTGA CACCGGAAGAAAGCTTGGGGCAATACCAATTGATTTGGTGAAAGGAACATCAGGCAGCCCCATTCTCAAC GCCCAGGGAGTGGTTGTGGGGCTATACGGAAATGGCCTAAAAACTAATGAGACCTACGTCAGCAGCATTG CTCAAGGGGAAGCGGAGAAGAGTCGCCCCAACCTCCCACAGGCTGTTGTGGGTACGGGCTGGACATCAAA GGGTCAGATCACAGTGCTGGACATGCACCCAGGCTCGGGGAAGACCCACAGAGTCCTCCCGGAGCTCATT CGCCAATGCATTGACAGGCGCCTGAGAACGTTGGTGTTGGCTCCAACTCGTGTGGTACTCAAAGAAATGG AGCGTGCTTTGAATGGGAAACGGGTCAGGTTCCACTCACCAGCAGTCAGTGACCAACAGGCTGGAGGGGC AATTGTCGATGTGATGTGTCACGCAACCTATGTCAACAGAAGGCTACTCCCACAGGGAAGACAAAATTGG GAGGTGGCAATCATGGATGAGGCCCACTGGACGGACCCCCACAGCATAGCTGCCAGAGGTCATTTGTATA CTCTGGCAAAAGAAAACAAGTGTGCACTGGTCTTGATGACAGCGACACCTCCTGGTAAGAGTGAACCCTT TCCGGAGTCCAATGGAGCCATTACTAGTGAGGAAAGACAGATTCCTGATGGGGAGTGGCGTGACGGGTTT GACTGGATCACTGAGTATGAAGGGCGCACCGCCTGGTTTGTCCCTTCGATTGCAAAAGGTGGTGCTATAG CTCGCACCTTGAGACAGAAGGGGAAAAGTGTGATTTGTTTGAACAGCAAAACCTTTGAAAAGGACTACTC CAGAGTGAGGGATGAGAAGCCTGACTTTGTGGTGACGACTGATATCTCGGAGATGGGAGCTAACCTTGAC GTGAGCCGCGTCATAGATGGGAGGACAAACATCAAGCCCGAGGAGGTTGATGGGAAAGTCGAGCTCACCG GGACCAGGCGAGTGACCACGGCTTCCGCTGCCCAACGGCGCGGAAGAGTTGGTCGGCAAGACGGACGAAC AGATGAATACATATACTCTGGACAGTGTGATGATGATGACAGTGGATTAGTGCAATGGAAAGAGGCGCAA ATACTTCTTGACAACATAACAACCTTGCGGGGGCCCGTGGCCACCTTCTATGGACCAGAACAGGACAAGA TGCCGGAGGTGGCCGGTCACTTTCGACTCACTGAAGAGAAAAGAAAGCACTTCCGACATCTTCTCACCCA TTGTGACTTCACACCGTGGCTGGCATGGCACGTCGCAGCGAATGTATCCAGCGTCACGGATCGAAGCTGG ACATGGGAAGGGCCGGAGGCAAATGCCGTGGATGAGGCCAGTGGTGACTTGGTCACCTTTAGGAGCCCGA ATGGGGCGGAGAGAACTCTCAGGCCGGTGTGGAAGGACGCACGTATGTTCAAAGAGGGACGTGACATCAA AGAGTTCGTGGCGTACGCGTCTGGGCGTCGCAGCTTCGGAGATGTTCTGACAGGAATGTCGGGAGTTCCT GAGCTCCTGCGGCACAGATGCGTCAGTGCCCTGGATGTCTTCTACACGCTTATGCATGAGGAACCTGGCA GCAGGGCAATGAGAATGGCGGAGAGAGATGCCCCAGAGGCCTTTCTGACTATGGTTGAGATGATGGTGCT GGGTTTGGCAACCCTGGGTGTCATCTGGTGCTTCGTCGTCCGGACTTCAATCAGCCGCATGATGCTGGGC ACGCTGGTCCTGCTGGCCTCCTTGCTACTCTTGTGGGCAGGTGGCGTCGGCTATGGGAACATGGCTGGAG TGGCTCTCATCTTTTACACGTTGCTGACGGTGCTGCAGCCTGAGGCGGGAAAACAGAGAAGCAGTGACGA CAACAAACTGGCATATTTCTTGCTGACGCTCTGCAGCCTTGCTGGACTGGTTGCAGCCAATGAGATGGGC TTTCTGGAGAAGACCAAGGCAGACTTGTCCACGGCGCTGTGGAGTGAACGGGAGGAACCCCGGCCATGGA GTGAATGGACGAATGTGGACATCCAGCCAGCGAGGTCCTGGGGGACCTATGTGCTGGTGGTGTCTCTGTT CACACCTTACATCATCCACCAACTGCAGACCAAAATCCAACAACTTGTCAACAGTGCCGTGGCATCTGGT GCACAGGCCATGAGAGACCTTGGGGGAGGTGCCCCCTTCTTTGGTGTGGCGGGACATGTCATGACCCTCG GGGTGGTGTCACTGATTGGGGCTACTCCCACCTCACTGATGGTGGGCGTTGGCTTGGCGGCACTCCATCT GGCCATTGTGGTGTCTGGTCTGGAGGCTGAATTAACACAGAGAGCTCATAAGGTCTTTTTCTCTGCAATG GTGCGCAACCCCATGGTGGATGGGGATGTCATCAACCCATTCGGGGAGGGGGAGGCAAAACCTGCTCTAT ATGAAAGGAAAATGAGTCTGGTGTTGGCCACAGTGTTGTGCCTCATGTCGGTGGTCATGAACCGAACGGT GGCCTCCATAACAGAGGCTTCAGCAGTGGGACTGGCAGCAGCGGGACAGCTGCTTAGACCGGAGGCTGAC ACGTTGTGGACGATGCCGGTTGCTTGTGGCATGAGTGGTGTGGTCAGGGGTAGCCTGTGGGGGTTTTTGC CTCTTGGGCATAGACTCTGGCTTCGAGCCTCTGGGGGTAGGCGTGGTGGTTCTGAGGGAGACACGCTTGG AGATCTCTGGAAGCGGAGGCTGAACAACTGCACGAGGGAGGAATTCTTTGTGTACAGGCGCACCGGCATC CTGGAGACGGAACGTGACAAGGCTAGAGAGTTGCTCAGAAGAGGAGAGACCAATGTGGGATTGGCTGTCT CTCGGGGCACGGCAAAGCTTGCCTGGCTTGAGGAACGCGGATATGCCACCCTCAAGGGAGAGGTGGTAGA TCTTGGATGTGGAAGGGGCGGCTGGTCCTATTATGCGGCATCCCGACCGGCAGTCATGAGTGTCAGGGCA TATACCATTGGTGGAAAAGGGCACGAGGCTCCAAAGATGGTAACAAGCCTGGGTTGGAACTTGATTAAAT TCAGATCAGGAATGGACGTGTTCAGCATGCAGCCACACCGGGCTGACACTGTCATGTGTGACATCGGAGA GAGCAGCCCAGATGCCGCTGTGGAGGGTGAGAGGACAAGGAAAGTGATACTGCTCATGGAGCAATGGAAA AACAGGAACCCCACGGCTGCCTGTGTGTTCAAGGTGCTGGCCCCATATCGCCCAGAAGTGATAGAGGCAC TGCACAGATTCCAACTGCAATGGGGGGGGGGTCTGGTGAGGACCCCTTTTTCAAGGAACTCCACCCATGA GATGTATTACTCAACAGCCGTCACTGGGAACATAGTGAACTCCGTCAATGTACAGTCGAGGAAACTTTTG GCTCGGTTTGGAGACCAGAGAGGGCCAACCAAGGTGCCTGAACTCGACCTGGGAGTTGGAACGAGGTGTG TGGTCTTAGCTGAGGACAAGGTGAAAGAACAAGACGTACAAGAGAGGATCAGAGCGTTGCGGGAGCAATA CAGCGAAACCTGGCATATGGACGAGGAACACCCGTACCGGACATGGCAGTACTGGGGCAGCTACCGCACG GCACCAACCGGCTCGGCGGCGTCACTGATCAATGGGGTTGTGAAACTTCTCAGCTGGCCATGGAACGCAC GGGAAGATGTGGTGCGCATGGCTATGACTGACACAACGGCTTTCGGACAGCAGAGAGTGTTCAAAGATAA AGTTGACACAAAGGCACAGGAGCCTCAGCCCGGTACAAGAGTCATCATGAGAGCTGTAAATGATTGGATT TTGGAACGACTGGCGCAGAAAAGCAAACCGCGCATGTGCAGCAGGGAAGAATTCATAGCAAAAGTGAAAT CAAATGCAGCCTTGGGAGCTTGGTCAGATGAGCAAAACAGATGGGCAAGTGCAAGAGAGGCTGTAGAGGA TCCTGCATTCTGGCGCCTCGTGGATGAAGAGAGAGAAAGGCACCTCATGGGGAGATGTGCGCACTGCGTG TACAACATGATGGGCAAGAGAGAAAAGAAACTGGGAGAGTTCGGAGTGGCGAAAGGAAGTCGGGCCATTT GGTACATGTGGCTGGGGAGTCGCTTTTTGGAGTTCGAGGCTCTTGGATTCTTGAATGAAGACCATTGGGC CTCTAGAGAGTCCAGTGGAGCTGGAGTTGAGGGAATAAGCTTGAACTACCTGGGCTGGCACCTCAAGAAG TTGTCAACCCTGAATGGAGGACTCTTCTATGCAGATGACACAGCTGGCTGGGACACGAAAGTTACCAATG CAGACTTAGAGGATGAAGAACAGATCCTACGGTACATGGAGGGTGAGCACAAACAATTGGCAACCACAAT AATGCAAAAAGCATACCATGCCAAAGTCGTGAAGGTCGCGAGGCCTTCCCGTGATGGAGGCTGCATCATG GATGTCATCACAAGAAGAGACCAAAGAGGTTCGGGTCAGGTTGTGACCTATGCCCTTAACACCCTCACCA ACATAAAGGTGCAATTAATCCGAATGATGGAAGGGGAAGGGGTCATAGAGGCAGCGGATGCACACAACCC GAGACTGCTTCGAGTGGAGCGCTGGCTGAAAGAACACGGAGAAGAGCGTCTTGGAAGAATGCTCGTCAGT GGTGACGATTGTGTGGTGAGGCCCTTGGATGACAGATTTGGCAAAGCACTTTACTTTCTGAATGACATGG CCAAGACCAGGAAGGACATTGGGGAATGGGAGCACTCAGCCGGCTTTTCAAGCTGGGAGGAGGTACCCTT TTGTTCACACCATTTCCACGAGCTAGTGATGAAGGATGGACGCACCCTGGTGGTGCCGTGCCGAGACCAA GATGAACTCGTTGGGAGGGCGCGCATCTCACCGGGGTGCGGCTGGAGTGTCCGCGAGACGGCCTGCCTTT CAAAAGCCTACGGGCAGATGTGGCTGCTGAGCTACTTCCACCGACGAGACCTGAGGACGCTCGGGCTTGC CATTAACTCAGCAGTGCCTGCCGATTGGGTTCCTACCGGCCGCACGACGTGGAGCATTCATGCCAGTGGG GCCTGGATGACCACAGAAGACATGCTGGACGTTTGGAACCGGGTGTGGATTCTGGACAACCCTTTCATGC AGAACAAGGAAAGGGTCATGGAGTGGAGGGATGTTCCGTACCTCCCTAAAGCTCAGGACATGTTATGTTC CTCCCTTGTTGGGAGGAGAGAAAGAGCAGAATGGGCCAAGAACATCTGGGGAGCGGTGGAAAAGGTGAGG AAGATGATAGGTCCTGAAAAGTTCAAGGACTATCTCTCCTGTATGGACCGCCATGACCTGCACTGGGAGC TCAGACTGGAGAGCTCAATAATCTA >NP_043135.1 polyprotein [Tick-borne encephalitis virus] MVKKAILKGKGGGPPRRVSKETATKTRQPRVQMPNGLVLMRMMGILWHAVAGTARNPVLKAFWNSVPLKQ ATAALRKIKRTVSALMVGLQKRGKRRSATDWMSWLLVITLLGMTLAATVRKERDGSTVIRAEGKDAATQV RVENGTCVILATDMGSWCDDSLSYECVTIDQGEEPVDVDCFCRNVDGVYLEYGRCGKQEGSRTRRSVLIP SHAQGELTGRGHKWLEGDSLRTHLTRVEGWVWKNKLLALAMVTVVWLTLESVVTRVAVLVVLLCLAPVYA SRCTHLENRDFVTGTQGTTRVTLVLELGGCVTITAEGKPSMDVWLDAIYQENPAKTREYCLHAKLSDTKV AARCPTMGPATLAEEHQGGTVCKRDQSDRGWGNHCGLFGKGSIVACVKAACEAKKKATGHVYDANKIVYT VKVEPHTGDYVAANETHSGRKTASFTISSEKTILTMGEYGDVSLLCRVASGVDLAQTVILELDKTVEHLP TAWQVHRDWFNDLALPWKHEGAQNWNNAERLVEFGAPHAVKMDVYNLGDQTGVLLKALAGVPVAHIEGTK YHLKSGHVTCEVGLEKLKMKGLTYTMCDKTKFTWKRAPTDSGHDTVVMEVTFSGTKPCRIPVRAVAHGSP DVNVAMLITPNPTIENNGGGFIEMQLPPGDNIIYVGELSHQWFQKGSSIGRVFQKTKKGIERLTVIGEHA WDFGSAGGFLSSIGKAVHTVLGGAFNSIFGGVGFLPKLLLGVALAWLGLNMRNPTMSMSFLLAGGLVLAM TLGVGADVGCAVDTERMELRCGEGLVVWREVSEWYDNYAYYPETPGALASAIKETFEEGSCGVVPQNRLE MAMWRSSVTELNLALAEGEANLTVVVDKFDPTDYRGGVPGLLKKGKDIKVSWKSWGHSMIWSIPEAPRRF MVGTEGQSECPLERRKTGVFTVAEFGVGLRTKVFLDFRQEPTHECDTGVMGAAVKNGMAIHTDQSLWMRS MKNDTGTYIVELLVTDLRNCSWPASHTIDNADVVDSELFLPASLAGPRSWYNRIPGYSEQVKGPWKYTPI RVIREECPGTTVTINAKCDKRGASVRSTTESGKVIPEWCCRACTMPPVTFRTGTDCWYAMEIRPVHDQGG LVRSMVVADNGELLSEGGVPGIVALFVVLEYIIRRRPSTGTTVVWGGIVVLALLVTGMVRIESLVRYVVA VGITFHLELGPEIVALMLLQAVFELRVGLLSAFALRRSLTVREMVTTYFLLLVLELGLPGASLEEFWKWG DALAMGALIFRACTAEGKTGAGLLLMALMTQQDVVTVHHGLVCFLSVASACSVWRLLKGHREQKGLTWVV PLAGLLGGEGSGIRLLAFWELSAHRGRRSFSEPLTVVGVMLTLASGMMRHTSQEALCALAVASFLLLMLV LGTRKMQLVAEWSGCVEWYPELVNEGGEVSLRVRQDAMGNFHLTELEKEERMMAFWLIAGLAASAIHWSG ILGVMGLWTLTEMLRSSRRSDLVFSGQGGRERGDRPFEVKDGVYRIFSPGLFWGQNQVGVGYGSKGVLHT MWHVTRGAALSIDDAVAGPYWADVREDVVCYGGAWSLEEKWKGETVQVHAFPPGRAHEVHQCQPGELILD TGRKLGAIPIDLVKGTSGSPILNAQGVVVGLYGNGLKTNETYVSSIAQGEAEKSRPNLPQAVVGTGWTSK GQITVLDMHPGSGKTHRVLPELIRQCIDRRLRTLVLAPTRVVLKEMERALNGKRVRFHSPAVSDQQAGGA IVDVMCHATYVNRRLLPQGRQNWEVAIMDEAHWTDPHSIAARGHLYTLAKENKCALVLMTATPPGKSEPF PESNGAITSEERQIPDGEWRDGFDWITEYEGRTAWFVPSIAKGGAIARTLRQKGKSVICLNSKTFEKDYS RVRDEKPDFVVTTDISEMGANLDVSRVIDGRTNIKPEEVDGKVELTGTRRVTTASAAQRRGRVGRQDGRT DEYIYSGQCDDDDSGLVQWKEAQILLDNITTLRGPVATFYGPEQDKMPEVAGHFRLTEEKRKHFRHLLTH CDFTPWLAWHVAANVSSVTDRSWTWEGPEANAVDEASGDLVTFRSPNGAERTLRPVWKDARMFKEGRDIK EFVAYASGRRSFGDVLTGMSGVPELLRHRCVSALDVFYTLMHEEPGSRAMRMAERDAPEAFLTMVEMMVL GLATLGVIWCFVVRTSISRMMLGTLVLLASLLLLWAGGVGYGNMAGVALIFYTLLTVLQPEAGKQRSSDD NKLAYFLLTLCSLAGLVAANEMGFLEKTKADLSTALWSEREEPRPWSEWTNVDIQPARSWGTYVLVVSLF TPYIIHQLQTKIQQLVNSAVASGAQAMRDLGGGAPFFGVAGHVMTLGVVSLIGATPTSLMVGVGLAALHL AIVVSGLEAELTQRAHKVFFSAMVRNPMVDGDVINPFGEGEAKPALYERKMSLVLATVLCLMSVVMNRTV ASITEASAVGLAAAGQLLRPEADTLWTMPVACGMSGVVRGSLWGFLPLGHRLWLRASGGRRGGSEGDTLG DLWKRRLNNCTREEFFVYRRTGILETERDKARELLRRGETNVGLAVSRGTAKLAWLEERGYATLKGEVVD LGCGRGGWSYYAASRPAVMSVRAYTIGGKGHEAPKMVTSLGWNLIKFRSGMDVFSMQPHRADTVMCDIGE SSPDAAVEGERTRKVILLMEQWKNRNPTAACVFKVLAPYRPEVIEALHRFQLQWGGGLVRTPFSRNSTHE MYYSTAVTGNIVNSVNVQSRKLLARFGDQRGPTKVPELDLGVGTRCVVLAEDKVKEQDVQERIRALREQY SETWHMDEEHPYRTWQYWGSYRTAPTGSAASLINGVVKLLSWPWNAREDVVRMAMTDTTAFGQQRVFKDK VDTKAQEPQPGTRVIMRAVNDWILERLAQKSKPRMCSREEFIAKVKSNAALGAWSDEQNRWASAREAVED PAFWRLVDEERERHLMGRCAHCVYNMMGKREKKLGEFGVAKGSRAIWYMWLGSRFLEFEALGFLNEDHWA SRESSGAGVEGISLNYLGWHLKKLSTLNGGLFYADDTAGWDTKVTNADLEDEEQILRYMEGEHKQLATTI MQKAYHAKVVKVARPSRDGGCIMDVITRRDQRGSGQVVTYALNTLTNIKVQLIRMMEGEGVIEAADAHNP RLLRVERWLKEHGEERLGRMLVSGDDCVVRPLDDRFGKALYFLNDMAKTRKDIGEWEHSAGFSSWEEVPF CSHHFHELVMKDGRTLVVPCRDQDELVGRARISPGCGWSVRETACLSKAYGQMWLLSYFHRRDLRTLGLA INSAVPADWVPTGRTTWSIHASGAWMTTEDMLDVWNRVWILDNPFMQNKERVMEWRDVPYLPKAQDMLCS SLVGRRERAEWAKNIWGAVEKVRKMIGPEKFKDYLSCMDRHDLHWELRLESSII Protective antigen Series of plasmid constructs encoding different forms of the envelope glycoprotein E of the flavivirus tick-borne encephalitis virus were constructed. These included a secreted recombinant subviral particle, a secreted carboxyl-terminally truncated soluble homodimer, a nonsecreted full-length form, and an inefficiently secreted truncated form. Mice were immunized using both i.m. injection and Gene Gun-mediated application of plasmids. The functional immune response was evaluated by determining specific neutralizing and hemagglutination-inhibiting Ab activities and by challenging the mice with a lethal dose of the virus.The plasmid construct encoding a secreted subviral particle, which carries multiple copies of the protective Ag on its surface, was superior to the other constructs in terms of extent and functionality of the Ab response as well as protection against virus challenge [Ref1249:Aberle et al., 1999]. Aberle et al., 1999 journal Aberle JH, Aberle SW, Allison SL, Stiasny K, Ecker M, Mandl CW, Berger R, Heinz FX 1999 163 12 6756-6761 Journal of immunology (Baltimore, Md. : 1950) CDC: TBE website http://www.cdc.gov/ncidod/dvrd/Spb/mnpages/dispages/TBE.htm Dmitriev et al., 1996 journal Dmitriev IP, Khromykh AA, Ignatyev GM, Gainullina MN, Ageenko VA, Dryga SA, Vorobyeva MS, Sandakhchiev LS 1996 44 1-3 97-103 Journal of biotechnology Kofler et al., 2002 journal Kofler RM, Heinz FX, Mandl CW 2002 76 7 3534-3543 Journal of virology Stenkova et al., 2017 journal Stenkova AM, Chopenko NS, Davydova LA, Mazeika AN, Bystritskaya EP, Portnyagina OY, Anastyuk SD, Kulbatskii DS, Lyukmanova EN, Dolgikh DA, Kostetsky EY, Sanina NM 2017 24 10 974-981 Protein and peptide letters