African horse sickness virus 40050 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 ALVAC-AHSV VO_0004725 Recombinant vector vaccine Research Intramuscular injection (i.m.) Recombinant canarypox virus vector (ALVAC-AHSV) co-expressing synthetic genes encoding the outer capsid proteins (VP2 and VP5) of AHSV serotype 4 (AHSV-4) (Guthrie et al., 2009). Intramuscular injection (i.m.) Two groups of four horses each (two males and two females) were inoculated intramuscularly with 10^7.1 or 10^6.4 TCID50, respectively, of ALVAC–AHSV in approximately 1 ml of diluent containing a Carbopol® 974P adjuvant (BF Goodrich) (Guthrie et al., 2009). VO_0003057 Post challenge, all horses remained healthy and showed no adverse effects after vaccination (Guthrie et al., 2009). All nine horses were challenged by intravenous inoculation of 105.5 TCID50 of AHSV-4 at 28 days after the second vaccination (week 8) (Guthrie et al., 2009). Plant Produced AHSV Serotype 5 VLPs Vaccine Subunit vaccine Research Intraperitoneal injection (i.p.) A plant-produced AHSV-1/5 VLP (Chimaeric Virus-Like Particles) provides protection against AHSV-5 in IFNAR -/- mice. (O'Kennedy et al., 2022) 5% Montanide GEL 01 PR, Seppic, France (O'Kennedy et al., 2022) AHSV-1/5 VLPs were produced in N. benthamiana 𝚫XT/FT. The AHSV virion is a triple layered particle formed by the outer capsids (VP2 and VP5), the middle layer (VP7), and the inner shell (VP3, subcore). D-(+)-Trehalose dihydrate (Sigma-Aldrich)(5 % m/v) was added to the TFF purified VLPs before filter sterilization with a 0.45 lM + 0.2 lM Sartopore 2 sterile capsule (Sartorius, 5441307H4) using a peristaltic pump. The appropriate filter sterilized VLPs were mixed with autoclaved adjuvant (5% Montanide GEL 01 PR, Seppic, France) immediately before vaccination. (O'Kennedy et al., 2022) Intraperitoneal injection (i.p.) Plant-produced AHSV-1/5 VLPs displaying VP2 and VP5 of serotype 5 as the outer shell lay- ered on the AHSV serotype 1 core (VP3/VP7) (O'Kennedy et al., 2022) Recombinant protein preparation Recombinant protein preparation Mice vaccinated with a single dose of plant-produced chimaeric AHSV- 1/5 VLPs seroconverted at a 5 µg and 10 µg vaccine dose, within the first 14 days. The primary vaccination of AHSV-1/5 VLPs led to seroconversion of 1:40 (1.6 log10) and 1:28 (1.45 log10) when vaccinated with 5 and 10 lg AHSV-1/5 VLPs already on day 14, respectively. A booster vaccine was however necessary to elevate the neutralizing antibodies (nAbs) to 1:320 (2.5 log10) on day 28 for all VLP vaccine doses. Ten micrograms of soluble VP2 per mouse were required to equal this immune response after prime-boost vaccination. All the test antigens indicated a measure of CD4+/ CD8+ stimulation which is required to induce cell memory. Mice vaccinated with plant-produced AHSV-1/5 VLPs showed a larger increase in stimulation as compared to the OBP BEI inactivated vaccine as positive control and to a lesser extent for the soluble VP2 vaccinated mice. (O'Kennedy et al., 2022) Eight groups of IFNAR-/- mice (n = 3) were vaccinated intraperitoneally with 1 µg, 5 µg or 10 µg plant-produced VLPs or VP2. Five groups of mice (n = 6 for vaccinated; and n = 3 for control groups) were used in the challenge study. Mice in the challenge study, were prime boost vaccinated (days 0 and 14) with either 10 µg VLPs (group 1) or 10 µg VP2 (group 2) or PBS buffer (negative control, group 3) or BEI inactivated 5x10^4 PFU (positive control, group 4). (O'Kennedy et al., 2022) Protection against AHSV-5 conferred by both plant-produced adjuvanted VLPs and VP2 vaccines correlated strongly with SNTs determined during the immunogenicity study and mice (n = 6) of each group survived until day 25 post challenge when the study was terminated. The negative control group (PBS with adjuvant) succumbed within 8– 11 days after challenge. (O'Kennedy et al., 2022) Groups 1–4 were challenged 28 days after the primary vaccine. Challenge with a dose containing 1.4 X 10^5 pfu of AHSV-5 per mouse on day 28 (14 days post booster immunization) was administered subcutaneously. (O'Kennedy et al., 2022) Plant Produced AHSV VP2 vaccine Subunit vaccine Research Intraperitoneal injection (i.p.) A plant produced AHSV Vaccine expressing VP2 antigen protects against AHSV-5 challenge in mice. (O'Kennedy et al., 2022) 5% Montanide GEL 01 PR, Seppic, France (O'Kennedy et al., 2022) AHSV-5 VP2 was produced in N. benthamiana DXT/FT. D-(+)-Trehalose dihydrate (Sigma-Aldrich)(5 % m/v) was added to the IMAC purified VP2 antigens before filter sterilization with a 0.45 lM + 0.2 lM Sartopore 2 sterile capsule (Sartorius, 5441307H4) using a peristaltic pump. The appropriate filter sterilised VP2 antigens were mixed with autoclaved adjuvant (5% Montanide GEL 01 PR, Seppic, France) immediately before vaccination. (O'Kennedy et al., 2022) Intraperitoneal injection (i.p.) VP2 (O'Kennedy et al., 2022) Recombinant protein preparation Mice vaccinated with a single dose of plant-produced chimaeric AHSV- 1/5 VLPs seroconverted at a 5 µg and 10 µg vaccine dose, within the first 14 days. The primary vaccination of AHSV-1/5 VLPs led to seroconversion of 1:40 (1.6 log10) and 1:28 (1.45 log10) when vaccinated with 5 and 10 lg AHSV-1/5 VLPs already on day 14, respectively. A booster vaccine was however necessary to elevate the neutralizing antibodies (nAbs) to 1:320 (2.5 log10) on day 28 for all VLP vaccine doses. Ten micrograms of soluble VP2 per mouse were required to equal this immune response after prime-boost vaccination. All the test antigens indicated a measure of CD4+/ CD8+ stimulation which is required to induce cell memory. Mice vaccinated with plant-produced AHSV-1/5 VLPs showed a larger increase in stimulation as compared to the OBP BEI inactivated vaccine as positive control and to a lesser extent for the soluble VP2 vaccinated mice. (O'Kennedy et al., 2022) Eight groups of IFNAR-/- mice (n = 3) were vaccinated intraperitoneally with 1 µg, 5 µg or 10 µg plant-produced VLPs or VP2. Five groups of mice (n = 6 for vaccinated; and n = 3 for control groups) were used in the challenge study. Mice in the challenge study, were prime boost vaccinated (days 0 and 14) with either 10 µg VLPs (group 1) or 10 µg VP2 (group 2) or PBS buffer (negative control, group 3) or BEI inactivated 5x104 PFU (positive control, group 4). (O'Kennedy et al., 2022) Protection against AHSV-5 conferred by both plant-produced adjuvanted VLPs and VP2 vaccines correlated strongly with SNTs determined during the immunogenicity study and mice (n = 6) of each group survived until day 25 post challenge when the study was terminated. The negative control group (PBS with adjuvant) succumbed within 8– 11 days after challenge. (O'Kennedy et al., 2022) Groups 1–4 were challenged 28 days after the primary vaccine. Challenge with a dose containing 1.4 X 105 pfu of AHSV-5 per mouse on day 28 (14 days post booster immunization) was administered subcutaneously. (O'Kennedy et al., 2022) rMVA- AHS-VP2 VO_0004758 Recombinant vector vaccine Research Intramuscular injection (i.m.) Recombinant vaccine based on modified vaccinia Ankara expressing AHS-4 VP2 (MVA-VP2) (Castillo-Olivares et al., 2011). Intramuscular injection (i.m.) The mice were vaccinated by the intra-peritoneal route on days 0 and 28 with 10^7 pfu of MVA-VP2 per mouse (Castillo-Olivares et al., 2011). VO_0000287 All of the MVA-VP2 vaccinated animals were also protected from clinical signs and were completely healthy until the end of the study period (Castillo-Olivares et al., 2011). The mice were subsequently challenged on day 35 with 106 pfu of AHSV-4 (Castillo-Olivares et al., 2011). VP2 African horse sickness virus CDJ98824 CDD:279269 40050 VP2 8.83 9039.572 141 Orbivirus outer capsid protein VP2; pfam00898 >CDJ98824.1 VP2, partial [African horse sickness virus] WVXWVVNYIMLSHVKRLVKDYKFKKLQPDNLMSGMNKLVGALRCYAYCLILALYDHFGAEIEGFRKGTNA ASIVETVSQMFP Protective antigen [Ref4957:Manning et al., 2017] VP7 African horse sickness virus AKP39983 CDD:279268 36421 VP7 6.27 34424.22 404 passage history: 1# BHK >AKP39983.1 VP7 [African horse sickness virus 4] MDAIAARALSVVRACVTVTDARVSLDPGVMETLGIAINRYNGLTNHSVSMRPQTQAERNEMFFMCTDMVL AALNVQIGNISPDYDQALATVGALATTEIPYNVQAMNDIVRITGQMQTFGPSKVQTGPYAGAVEVQQSGR YYVPQGRTRGGYINSNIAEVCMDAGAAGQVNALLAPRRGDAVMIYFVWRPLRIFCDPQGASLESAPGTFV TVDGVNVAAGDVVAWNTIAPVNVGNPGARRSILQFEVLWYTSLDRSLDTVPELAPTLTRCYAYVSPTWHA LRAVIFQQMNMQPINPPIFPPTERNEIVAYLLVASLADVYAALRPDFRMNGVVAPVGQINRALVLAAYH Protective antigen [Ref4954:Rutkowska et al., 2011] Castillo-Olivares et al., 2011 journal Castillo-Olivares J, Calvo-Pinilla E, Casanova I, Bachanek-Bankowska K, Chiam R, Maan S, Nieto JM, Ortego J, Mertens PP 2011 6 1 e16503 PloS one de et al., 2015 journal de la Poza F, Marín-López A, Castillo-Olivares J, Calvo-Pinilla E, Ortego J 2015 210 149-153 Virus research Guthrie et al., 2009 journal Guthrie AJ, Quan M, Lourens CW, Audonnet JC, Minke JM, Yao J, He L, Nordgren R, Gardner IA, Maclachlan NJ 2009 27 33 4434-4438 Vaccine Manning et al., 2017 journal Manning NM, Bachanek-Bankowska K, Mertens PPC, Castillo-Olivares J 2017 35 44 6024-6029 Vaccine Rutkowska et al., 2011 journal Rutkowska DA, Meyer QC, Maree F, Vosloo W, Fick W, Huismans H 2011 156 1-2 35-48 Virus research