Japanese encephalitis virus is a virus from the family Flaviviridae. It causes the mosquito-borne Japanese encephalitis. Japanese encephalitis is the leading cause of viral encephalitis in Asia, with 30,000–50,000 cases reported annually. Case-fatality rates range from 0.3% to 60% and depends on the population and on age. Rare outbreaks in U.S. territories in Western Pacific have occurred. This disease often occurs in rural areas but not usually in urban areas. Severe rigors, fever, headache and malaise are non-specific symptoms for the first week. Signs which develop during the acute encephalitic stage include neck rigidity, cachexia, hemiparesis, convulsions, and fever. Mental retardation developed from this disease usually leads to coma. Mortality of this disease varies but is generally much higher in children (Wiki: Japanese encephalitis).
4. Microbial Pathogenesis
Japanese encephalitis virus infects the lumen of the endoplasmic reticulum and rapidly accumulates substantial amounts of viral proteins.
5. Host Ranges and Animal Models
Domestic pigs and wild birds are reservoirs of the virus; transmission to humans may cause severe symptoms.
6. Host Protective Immunity
Infection with JEV confers life-long immunity (Wiki: Japanese encephalitis). For JE, both arms have been considered to be important for protection from the disease, since passive transfer of neutralizing antibodies or virus-specific T cells including CTLs can protect mice from a lethal challenge (Konishi et al., 2003).
II. Vaccine Related Pathogen Genes
1. E
Gene Name :
E
Sequence Strain (Species/Organism) :
Japanese encephalitis virus
Molecule Role Annotation :
Researchers constructed a recombinant pseudotype baculovirus encoding the JEV envelope (E) protein and demonstrated that it could elicit high protective immunity in mice. Intramuscular injections of BV-G-E at various doses into mice produced higher levels of JEV-specific neutralizing antibodies, IFN-gamma and better protective efficacy against a lethal challenge with JEV than that of pc-E. Furthermore, BV-G-E could elicit a higher level of cellular immunity response and provide equal protective efficacy against JEV challenge compared to inactivated vaccine (Li et al., 2009).
Molecule Role Annotation :
Balb/c mice and swine vaccinated with TK(-)/gG(-)/NS1(+) expressing NS1 protein of JEV could confer protective immunity against lethal challenge of the virulent PRV Ea strain and develop a good humoral and cellular immune response against JEV (Xu et al., 2004).
Molecule Role Annotation :
Researchers constructed plasmid DNAs encoding JE virus proteins. Cytotoxic T lymphocytes (CTLs) were induced by NS3 in a mouse model. Three immunizations with pUJENS3 provided a 50% partial protection from a lethal dose of JE virus (Konishi et al., 2003).
Molecule Role Annotation :
Researchers constructed plasmid DNAs encoding JE virus proteins. Cytotoxic T lymphocytes (CTLs) were induced by NS5 in a mouse model. Three immunizations with pUJENS5 provided a 57% partial protection from a lethal dose of JE virus (Konishi et al., 2003).
Molecule Role Annotation :
A cDNA clone representing the genome of structural proteins of Japanese encephalitis virus (JEV) was inserted into the thymidine kinase gene of vaccinia virus strains LC16mO and WR. Inoculation of rabbits with the infectious recombinant vaccinia virus resulted in rapid production of antiserum specific for the PreM and E glycoproteins of JEV. Mice vaccinated with the recombinant produced JEV-neutralizing antibodies and were resistant to challenge with JEV (Yasuda et al., 1990).
10. premembrane protein
Gene Name :
premembrane protein
Sequence Strain (Species/Organism) :
Japanese encephalitis virus
Molecule Role Annotation :
Researchers established a simple and effective method for DNA immunization against Japanese encephalitis virus (JEV) infection with plasmids encoding the viral PrM and E proteins and colloidal gold. After being inoculated twice, BALB/c mice were found to resist challenge with 100,000 times the 50% lethal dose (LD(50)) of JEV (Beijing-1 strain) even when immunized with a relatively small dose of 0.5 micro g of plasmid DNA (Zhao et al., 2003).
>gi|158516890|ref|YP_001527879.1| unnamed protein product [West Nile virus]
VTLSNFQGKVMMTVNATDVTDVITIPTAAGKNLCIVRAMDVGYMCDDTITYECPVLSAGNDPEDIDCWCT
KSAVYVRYGRCTKTRHSRRSRRSLTVQTHGESTLANKKGAWMDSTKATRYLVKTESWILRNPGYALVAAV
IGWMLGSNTMQRVVFVVLLLLVAPAYS
Molecule Role Annotation :
IFN-gamma plays a critical role in Th1 type immune response. It is important for protection against infections by various viruses and intracellular bacteria.
Additional Molecule Role :
Vaximmutor
Additional Molecule Role Annotation :
The experimental data demonstrated that three time vaccinations with BCG in BALB/c mice induced strong TB Ag-specific IFN-gamma immune responses in splenocytes (Wang et al., 2009).
Poxvirus-vectored vaccines for Japanese encephalitis (JE), NYVAC-JEV and ALVAC-JEV(Raengsakulrach et al., 1999).
f. Immunization Route
Intramuscular injection (i.m.)
g.
Monkey Response
Vaccination Protocol:
The vaccines were given to four monkeys each on study days 0 and 28 along with saline placebo on day 7. For controls, the licensed BIKEN JE vaccine and a saline placebo were given to other groups of four monkeys on days 0, 7, and 28 (Raengsakulrach et al., 1999).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
Two months after the booster dose, all 16 monkeys were challenged intranasally with one 90% effective dose of JEV strain KE-93 (AP61-1, C6/36-1, Mm-1, SM-2) (Raengsakulrach et al., 1999).
Efficacy:
This study suggests that the NYVAC-JEV and ALVAC-JEV vaccines are safe and immunogenic in monkeys and that the NYVAC-JEV and BIKEN vaccines are effective in protecting monkeys from encephalitis (Raengsakulrach et al., 1999).
2. Ixiaro
a. Product Name:
Japanese Encephalitis Vaccine, Inactivated, Adsorbed
JEV strain SA14-14-2 is propagated in Vero cells, and harvested. The harvested virus suspension is treated with protamine sulfate to remove contaminating DNA and proteins and centrifuged. Virus is inactivated by formaldehyde and aluminum hydroxide is added Ref917:FDA: Ixiaro].
l. Immunization Route
Intramuscular injection (i.m.)
m. Storage
Store in a refrigerator at 2° to 8° C (35° to 46° F). Do not freeze.
n . Approved Age for Licensed Use
2 months of age and older
3. Japanese encephalitis virus (JEV)
a. Type:
Recombinant vector vaccine
b. Status:
Licensed
c. Host Species for Licensed Use:
Baboon
d. Immunization Route
Intramuscular injection (i.m.)
4. Japanese encephalitis virus DNA vaccine encoding E protein
Description:
Plasmid pSLKJ12 contains the premembrane signal sequence as well as the premembrane (PrM) and envelope (E) genes of the Sagayama strain of JEV. The viral sequence, spanning nucleotides 408 to 2477, was retrieved from this plasmid by PstI-EcoRI digestion. The fragment was cloned into the eukaryotic expression vectors pcDL-SRα296 and pCAGGS, generously provided by Y. Takebe (Laboratory of Molecular Virology and Epidemiology, AIDS Research Center, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan) and J. Miyazaki (Division of Stem Cell Regulation Research, G6, Osaka University Medical School, Suita, Osaka, Japan), respectively. They are purified and designated pSRαJ12 and pCAGJ12, respectively (Zhao et al., 2003).
Description:
Plasmid pSLKJ12 contains the premembrane signal sequence as well as the premembrane (PrM) and envelope (E) genes of the Sagayama strain of JEV. The viral sequence, spanning nucleotides 408 to 2477, was retrieved from this plasmid by PstI-EcoRI digestion. The fragment was cloned into the eukaryotic expression vectors pcDL-SRα296 and pCAGGS, generously provided by Y. Takebe (Laboratory of Molecular Virology and Epidemiology, AIDS Research Center, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan) and J. Miyazaki (Division of Stem Cell Regulation Research, G6, Osaka University Medical School, Suita, Osaka, Japan), respectively (Zhao et al., 2003).
Vaccination Protocol:
For the protection test, 3-week-old female BALB/c mice were inoculated with either 50, 5, or 0.5 μg of the mixture on days 0 and 9 intravenously or intradermally (Zhao et al., 2003).
Challenge Protocol:
On day 22, all immunized mice were challenged by an intraperitoneal injection of 100,000 times the LD50 of JEV (Beijing-1 strain, 0.15 ml), at which time they were simultaneously inoculated intracerebrally with 25 μl of saline into the right hemisphere of their brains with a 27-gauge one-stop needle (Top Injection Needle, Tokyo, Japan) (Zhao et al., 2003).
Efficacy:
Researchers established a simple and effective method for DNA immunization against Japanese encephalitis virus (JEV) infection with plasmids encoding the viral PrM and E proteins and colloidal gold. After being inoculated twice, BALB/c mice were found to resist challenge with 100,000 times the 50% lethal dose (LD(50)) of JEV (Beijing-1 strain) even when immunized with a relatively small dose of 0.5 micro g of plasmid DNA (Zhao et al., 2003).
5. Japanese encephalitis virus DNA vaccine encoding NS1 (pUSK-NS1)
Description:
NS1 gene of Japanese encephalitis virus (JEV) SA14-14-2 strain was produced by reverse transcriptase-mediated PCR (RT-PCR) and was cloned into vector pUSK to form recombinant plasmid (designed as pUSK-NS1) (Xu et al., 2004).
Vaccination Protocol:
Ten 6-week-old Balb/c mice and eight 2-days old piglets were immunized with 5.0 log10pfu of the TK−/gG−/NS1+ mutant, while another equal number of animal in control groups with PBS (Xu et al., 2004).
Challenge Protocol:
The challenge was performed with PRV Ea strain at a concentration of 6.5 log10pfu (LD50=10−4.5) at day 7 for mice and day 14 for piglets post-vaccination (Xu et al., 2004).
Efficacy:
Balb/c mice and swine vaccinated with TK(-)/gG(-)/NS1(+) expressing NS1 protein of JEV could confer protective immunity against lethal challenge of the virulent PRV Ea strain and develop a good humoral and cellular immune response against JEV (Xu et al., 2004).
6. Japanese encephalitis virus DNA vaccine encoding PrM
Description:
Plasmid pSLKJ12 contains the premembrane signal sequence as well as the premembrane (PrM) and envelope (E) genes of the Sagayama strain of JEV. The viral sequence, spanning nucleotides 408 to 2477, was retrieved from this plasmid by PstI-EcoRI digestion. The fragment was cloned into the eukaryotic expression vectors pcDL-SRα296 and pCAGGS, generously provided by Y. Takebe (Laboratory of Molecular Virology and Epidemiology, AIDS Research Center, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan) and J. Miyazaki (Division of Stem Cell Regulation Research, G6, Osaka University Medical School, Suita, Osaka, Japan), respectively. They are purified and designated pSRαJ12 and pCAGJ12, respectively (Zhao et al., 2003).
Vaccination Protocol:
For the protection test, 3-week-old female BALB/c mice were inoculated with either 50, 5, or 0.5 μg of the mixture on days 0 and 9 intravenously or intradermally (Zhao et al., 2003).
Challenge Protocol:
On day 22, all immunized mice were challenged by an intraperitoneal injection of 100,000 times the LD50 of JEV (Beijing-1 strain, 0.15 ml), at which time they were simultaneously inoculated intracerebrally with 25 μl of saline into the right hemisphere of their brains with a 27-gauge one-stop needle (Top Injection Needle, Tokyo, Japan) (Zhao et al., 2003).
Efficacy:
Researchers established a simple and effective method for DNA immunization against Japanese encephalitis virus (JEV) infection with plasmids encoding the viral PrM and E proteins and colloidal gold. After being inoculated twice, BALB/c mice were found to resist challenge with 100,000 times the 50% lethal dose (LD(50)) of JEV (Beijing-1 strain) even when immunized with a relatively small dose of 0.5 micro g of plasmid DNA (Zhao et al., 2003).
Vaccination Protocol:
Control mice received 1 μg of pcDNA3.1/V5-His plasmid per dose. While same amount of recombinant plasmid, pcDNA3.1/V5-His-P-JEV was used to immunize the experimental group. Groups of BALB/c mice (n = 10), received two booster doses with equal amount of DNA after every 2 weeks (Kulkarni et al., 2012).
Vaccine Immune Response Type:
VO_0003057
Immune Response:
Anti-JEV antibodies were detectable in all mouse groups 31 days after immunization. Levels of these antibodies increased after the first booster immunization and were further enhanced after the second booster dose. Following the challenge, pcDNA3.1/V5-His-P-JEV immunized mice showed significantly enhanced JEV antibody titres (Kulkarni et al., 2012).
Challenge Protocol:
At 6 weeks after immunization, mice were challenged with lethal dose of 100LD50 with JEV 733913 strain by intraperitonial route, followed by 1% starch by the intracerebral route to breach the blood brain barrier (Kulkarni et al., 2012).
Efficacy:
Approximately 83% of mice immunized with the vaccine survived the challenge injection (Kulkarni et al., 2012).
8. Japanese encephalitis virus DNA Vaccine pCJ-3/E encoding E
Vaccination Protocol:
All mice were immunized intramuscularly at 6 to 8 weeks of age. Groups of five mice were anesthetized and injected three times at 3-week intervals with 100 μg of DNA (Wu et al., 2006).
Vaccine Immune Response Type:
VO_0003057
Immune Response:
The vaccine enhanced the production of higher titers of neutralizing antibodies, and generated both cellular and humoral immunity (Wu et al., 2006).
Challenge Protocol:
For a lethal challenge experiment, C3H/HeN mice were injected intraperitoneally with 50 times the LD50 of JEV Beijing-1 and intracerebrally with PBS (Wu et al., 2006).
Efficacy:
In terms of survival rates of immunized mice challenged with JEV virus at week 16, 100% protection was seen with pCJ-3/E-immunized mice (Wu et al., 2006).
9. Japanese encephalitis virus DNA vaccine pUJENS3
Japanese encephalitis virus non-structural protein NS3
e. Gene Engineering of
NS3
Type:
DNA vaccine construction
Description:
Amplified DNA fragments containing the NS3 gene was ligated to the pUBIQ vector at the EcoRI/XbaI site to construct pUJENS3 (Konishi et al., 2003).
Vaccination Protocol:
Four-to-six-week-old male BALB/c mice were immunized intramuscularly (i.m.) with 100 μg of plasmid DNA one to three times at intervals of 2 weeks. For evaluation of CTL induction, spleens were collected from groups of two immunized mice 2–4 weeks after the last immunization (Konishi et al., 2003).
Challenge Protocol:
For evaluation of protective efficacy, groups of 6–12 immunized mice were bled retroorbitally for evaluating pre-challenge serum neutralizing antibody titers, and then challenged i.p. with 100 or 400 LD50 of the P3 strain of JE virus, 6 weeks after the first immunization. Mice were observed for 21 days, and surviving mice were bled for evaluating post-challenge serum neutralizing antibody titers. For monitoring virus load following challenge, groups of 18 immunized mice were challenged with 100 LD50 of the P3 strain, and 600–700 μl of blood and the whole brain were collected 1 h (day 0) or consecutive 5 days (days 1–5) following challenge from 3 mice per day per group (Konishi et al., 2003).
Efficacy:
Researchers constructed plasmid DNAs encoding JE virus proteins. Cytotoxic T lymphocytes (CTLs) were induced by NS3 in a mouse model. Three immunizations with pUJENS3 provided a 50% partial protection from a lethal dose of JE virus (Konishi et al., 2003).
10. Japanese encephalitis virus DNA vaccine pUJENS5
Description:
Amplified DNA fragments containing the NS5 gene was ligated to the pUBIQ8 vector at the BspEI and/or XbaI sites to construct pUJENS5 (Konishi et al., 2003).
Vaccination Protocol:
Four-to-six-week-old male BALB/c mice were immunized intramuscularly (i.m.) with 100 μg of plasmid DNA one to three times at intervals of 2 weeks. For evaluation of CTL induction, spleens were collected from groups of two immunized mice 2–4 weeks after the last immunization (Konishi et al., 2003).
Challenge Protocol:
For evaluation of protective efficacy, groups of 6–12 immunized mice were bled retroorbitally for evaluating pre-challenge serum neutralizing antibody titers, and then challenged i.p. with 100 or 400 LD50 of the P3 strain of JE virus, 6 weeks after the first immunization. Mice were observed for 21 days, and surviving mice were bled for evaluating post-challenge serum neutralizing antibody titers. For monitoring virus load following challenge, groups of 18 immunized mice were challenged with 100 LD50 of the P3 strain, and 600–700 μl of blood and the whole brain were collected 1 h (day 0) or consecutive 5 days (days 1–5) following challenge from 3 mice per day per group (Konishi et al., 2003).
Efficacy:
Researchers constructed plasmid DNAs encoding JE virus proteins. Cytotoxic T lymphocytes (CTLs) were induced by NS5 in a mouse model. Three immunizations with pUJENS5 provided a 57% partial protection from a lethal dose of JE virus (Konishi et al., 2003).
Description:
Recombinant pseudotype baculovirus (BV-G-E) was generated by inserting JEV E gene fragment into pFastBac-VSV/G vector. BALB/c mice were immunized with BV-G-E and challenged with JEV wild-type strain. The neutralization antibody, interferon (IFN)- expression and release, and survival rate were analysed and compared with the group of immunized with inactivated vaccine and DNA vaccine (pc-E) encoding the same gene of JEV (Li et al., 2009).
baculovirus pseudotyped with vesicular stomatitis virus glycoprotein (VSVG)
g. Immunization Route
Intramuscular injection (i.m.)
h.
Mouse Response
Host Strain:
BALB/c
Vaccination Protocol:
Four-week old female BALB/c mice (purchased from the Animal Centre, Institute of Medicine (Hubei Province, China) were randomly divided into seven groups (15 mice per group). Three groups were injected intramuscularly (i.m.) with 100 µl of PBS containing 1 × 10^8, 1 × 10^9 and 1 × 10^10 PFU of BV-G-E, respectively. The other three groups were injected intramuscularly with 100 µl of PBS containing 1 × 10^10 PFU of BV-G-EGFP or 100 µg of pc-E (a DNA vaccine construct expressing E protein) or 100 µl inactivated vaccine. The last group was used as a negative control by intramuscularly injecting 100 µl of PBS. Booster immunizations were identically performed 3 weeks later (Li et al., 2009).
Challenge Protocol:
A subset of immunized mice was intraperitoneally (i.p.) challenged with 105 PFU of wild-type JEV P3 in 0.1 ml at 6 weeks following the initial immunization. Mouse mortality was monitored daily for 3 weeks (Li et al., 2009).
Efficacy:
Researchers constructed a recombinant pseudotype baculovirus encoding the JEV envelope (E) protein and demonstrated that it could elicit high protective immunity in mice. Intramuscular injections of BV-G-E at various doses into mice produced higher levels of JEV-specific neutralizing antibodies, IFN-gamma and better protective efficacy against a lethal challenge with JEV than that of pc-E. Furthermore, BV-G-E could elicit a higher level of cellular immunity response and provide equal protective efficacy against JEV challenge compared to inactivated vaccine (Li et al., 2009).
Host Gene Response of
Ifng (Interferon gamma)
Gene Response:
An evident dose-dependent pattern of IFN-gamma production could be observed in mice immunized with BV-G-E. As expected, no significant production of IFN-γ was detected in PBS-inoculated mice. Interestingly, the mean relative IFN-γ mRNA expression in the group of BV-G-EGFP was also significantly higher than that of pc-E vaccinated mice in splenocytes (Li et al., 2009).
Poxvirus-vectored vaccines for Japanese encephalitis (JE), NYVAC-JEV and ALVAC-JEV(Raengsakulrach et al., 1999).
f. Immunization Route
Intramuscular injection (i.m.)
g.
Monkey Response
Vaccination Protocol:
The vaccines were given to four monkeys each on study days 0 and 28 along with saline placebo on day 7. For controls, the licensed BIKEN JE vaccine and a saline placebo were given to other groups of four monkeys on days 0, 7, and 28 (Raengsakulrach et al., 1999).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
Two months after the booster dose, all 16 monkeys were challenged intranasally with one 90% effective dose of JEV strain KE-93 (AP61-1, C6/36-1, Mm-1, SM-2) (Raengsakulrach et al., 1999).
Efficacy:
This study suggests that the NYVAC-JEV and ALVAC-JEV vaccines are safe and immunogenic in monkeys and that the NYVAC-JEV and BIKEN vaccines are effective in protecting monkeys from encephalitis (Raengsakulrach et al., 1999).
Description:
A highly attenuated strain of vaccinia virus (NYVAC) was engineered to express the Japanese encephalitis virus (JEV) prM, E, and NS1 genes or the prM and E genes (Konishi et al., 1992).
Description:
A highly attenuated strain of vaccinia virus (NYVAC) was engineered to express the Japanese encephalitis virus (JEV) prM, E, and NS1 genes or the prM and E genes (Konishi et al., 1992).
Description:
A highly attenuated strain of vaccinia virus (NYVAC) was engineered to express the Japanese encephalitis virus (JEV) prM, E, and NS1 genes or the prM and E genes (Konishi et al., 1992).
Highly attenuated strain of vaccinia virus (NYVAC) was engineered to express the Japanese encephalitis virus (JEV) prM, E, and NS1 genes or the prM and E genes (Konishi et al., 1992).
i. Immunization Route
Intramuscular injection (i.m.)
j.
Pig Response
Vaccination Protocol:
The recombinant viruses were tested as vaccine candidates in pigs, a natural host of JEV. JEV-neutralizing and hemagglutination-inhibiting antibodies appeared in swine sera 7 days after immunization with 108 PFU of the recombinant viruses and increased after a second dose at 28 days (Konishi et al., 1992).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
JEV challenge (d56) of the swine with 2 × 10^8 PFU of JEV (Konishi et al., 1992).
Efficacy:
The JEV levels detected in the serum after JEV challenge were significantly reduced in animals inoculated with the recombinant viruses (Konishi et al., 1992).
3. Hua et al., 2014: Hua RH, Liu LK, Huo H, Li YN, Guo LP, Wang XL, Qin CF, Bu ZG. Comprehensive mapping of a novel NS1 epitope conserved in flaviviruses within the Japanese encephalitis virus serocomplex. Virus research. 2014; 185; 103-109. [PubMed: 24631788].
4. Konishi et al., 1992: Konishi E, Pincus S, Paoletti E, Laegreid WW, Shope RE, Mason PW. A highly attenuated host range-restricted vaccinia virus strain, NYVAC, encoding the prM, E, and NS1 genes of Japanese encephalitis virus prevents JEV viremia in swine. Virology. 1992; 190(1); 454-458. [PubMed: 1326813].
5. Konishi et al., 2003: Konishi E, Ajiro N, Nukuzuma C, Mason PW, Kurane I. Comparison of protective efficacies of plasmid DNAs encoding Japanese encephalitis virus proteins that induce neutralizing antibody or cytotoxic T lymphocytes in mice. Vaccine. 2003; 21(25-26); 3675-3683. [PubMed: 12922097].
6. Kulkarni et al., 2012: Kulkarni R, Sapkal G, Gore M. Evaluation of Japanese encephalitis virus polytope DNA vaccine candidate in BALB/c mice. Virus research. 2012; 170(1-2); 118-125. [PubMed: 23007095].
7. Li et al., 2009: Li Y, Ye J, Cao S, Xiao S, Zhao Q, Liu X, Jin M, Chen H. Immunization with pseudotype baculovirus expressing envelope protein of Japanese encephalitis virus elicits protective immunity in mice. The journal of gene medicine. 2009; 11(1); 57-65. [PubMed: 19006097].
8. Qian et al., 2015: Qian P, Zhi X, Wang B, Zhang H, Chen H, Li X. Construction and immune efficacy of recombinant pseudorabies virus expressing PrM-E proteins of Japanese encephalitis virus genotype ?. Virology journal. 2015; 12; 214. [PubMed: 26651827].
9. Raengsakulrach et al., 1999: Raengsakulrach B, Nisalak A, Gettayacamin M, Thirawuth V, Young GD, Myint KS, Ferguson LM, Hoke CH Jr, Innis BL, Vaughn DW. Safety, immunogenicity, and protective efficacy of NYVAC-JEV and ALVAC-JEV recombinant Japanese encephalitis vaccines in rhesus monkeys. The American journal of tropical medicine and hygiene. 1999; 60(3); 343-349. [PubMed: 10466959].
10. Sheets et al., 2006: Sheets RL, Stein J, Manetz TS, Duffy C, Nason M, Andrews C, Kong WP, Nabel GJ, Gomez PL. Biodistribution of DNA plasmid vaccines against HIV-1, Ebola, Severe Acute Respiratory Syndrome, or West Nile virus is similar, without integration, despite differing plasmid backbones or gene inserts. Toxicological sciences : an official journal of the Society of Toxicology. 2006; 91(2); 610-619. [PubMed: 16569729].
12. Wu et al., 2006: Wu CJ, Li TL, Huang HW, Tao MH, Chan YL. Development of an effective Japanese encephalitis virus-specific DNA vaccine. Microbes and infection / Institut Pasteur. 2006; 8(11); 2578-2586. [PubMed: 16949850].
13. Xu et al., 2004: Xu G, Xu X, Li Z, He Q, Wu B, Sun S, Chen H. Construction of recombinant pseudorabies virus expressing NS1 protein of Japanese encephalitis (SA14-14-2) virus and its safety and immunogenicity. Vaccine. 2004; 22(15-16); 1846-1853. [PubMed: 15121294].
14. Yasuda et al., 1990: Yasuda A, Kimura-Kuroda J, Ogimoto M, Miyamoto M, Sata T, Sato T, Takamura C, Kurata T, Kojima A, Yasui K. Induction of protective immunity in animals vaccinated with recombinant vaccinia viruses that express PreM and E glycoproteins of Japanese encephalitis virus. Journal of virology. 1990; 64(6); 2788-2795. [PubMed: 2159544].
15. Zhao et al., 2003: Zhao Z, Wakita T, Yasui K. Inoculation of plasmids encoding Japanese encephalitis virus PrM-E proteins with colloidal gold elicits a protective immune response in BALB/c mice. Journal of virology. 2003; 77(7); 4248-4260. [PubMed: 12634382].
