Abstract
Dengue is a major infectious disease that affects people living in tropical and subtropical regions around the world. The causative agents are dengue virus serotype 1, 2, 3, and 4 (DENV1, 2, 3, and 4). Developing a vaccine for dengue is a high priority for public health, but traditional methods have faced numerous obstacles due to the unique immunopathogenesis of dengue virus infection. Here, we report a novel dengue vaccine candidate based on dengue pseudoinfectious virus (PIV) produced by the incorporation of a dengue subgenomic replicon into viral particles in highly efficient packaging cells. The subgenomic replicon was constructed by deleting the capsid protein (C) gene from the dengue viral genome and optimizing the signal peptide sequence of pre-membrane protein (prM) to facilitate the formation of viral particles. Packaging cells were developed for inducible expression of a bi-protein Cpr, where the protein pr is the "pr" segment of viral protein prM that holds the protein C on the endoplasmic reticulum (ER). When the replicon was introduced into the packaging cells, protein C was released from the bi-protein Cpr by a replicon-encoded viral protease. Coordinate expression of viral structural proteins by the replicon and packaging cells led to the incorporation of the replicon into viral particle to produce PIVs. Animal tests showed that the dengue PIV vaccine was highly immunogenic and the immune response protected mice challenged with a hundred-fold LD50 inoculation of dengue virus. The method described here has the potential to be applied to vaccine development for other flaviviruses.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3854-3860 |
| Number of pages | 7 |
| Journal | Vaccine |
| Volume | 32 |
| Issue number | 30 |
| DOIs | |
| State | Published - 2014 |
| Externally published | Yes |
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Keywords
- Dengue vaccine
- Immunogenicity
- Packaging cell
- Pseudoinfectious virus
- Replicon
ASJC Scopus subject areas
- Molecular Medicine
- Immunology and Microbiology(all)
- veterinary(all)
- Public Health, Environmental and Occupational Health
- Infectious Diseases
Cite this
Highly efficient production of a dengue pseudoinfectious virus. / Pang, Xiaowu; Guo, Yinhan; Zhou, Yanfei; Fu, Wenchuan; Gu, Xinbin.
In: Vaccine, Vol. 32, No. 30, 2014, p. 3854-3860.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Highly efficient production of a dengue pseudoinfectious virus
AU - Pang, Xiaowu
AU - Guo, Yinhan
AU - Zhou, Yanfei
AU - Fu, Wenchuan
AU - Gu, Xinbin
PY - 2014
Y1 - 2014
N2 - Dengue is a major infectious disease that affects people living in tropical and subtropical regions around the world. The causative agents are dengue virus serotype 1, 2, 3, and 4 (DENV1, 2, 3, and 4). Developing a vaccine for dengue is a high priority for public health, but traditional methods have faced numerous obstacles due to the unique immunopathogenesis of dengue virus infection. Here, we report a novel dengue vaccine candidate based on dengue pseudoinfectious virus (PIV) produced by the incorporation of a dengue subgenomic replicon into viral particles in highly efficient packaging cells. The subgenomic replicon was constructed by deleting the capsid protein (C) gene from the dengue viral genome and optimizing the signal peptide sequence of pre-membrane protein (prM) to facilitate the formation of viral particles. Packaging cells were developed for inducible expression of a bi-protein Cpr, where the protein pr is the "pr" segment of viral protein prM that holds the protein C on the endoplasmic reticulum (ER). When the replicon was introduced into the packaging cells, protein C was released from the bi-protein Cpr by a replicon-encoded viral protease. Coordinate expression of viral structural proteins by the replicon and packaging cells led to the incorporation of the replicon into viral particle to produce PIVs. Animal tests showed that the dengue PIV vaccine was highly immunogenic and the immune response protected mice challenged with a hundred-fold LD50 inoculation of dengue virus. The method described here has the potential to be applied to vaccine development for other flaviviruses.
AB - Dengue is a major infectious disease that affects people living in tropical and subtropical regions around the world. The causative agents are dengue virus serotype 1, 2, 3, and 4 (DENV1, 2, 3, and 4). Developing a vaccine for dengue is a high priority for public health, but traditional methods have faced numerous obstacles due to the unique immunopathogenesis of dengue virus infection. Here, we report a novel dengue vaccine candidate based on dengue pseudoinfectious virus (PIV) produced by the incorporation of a dengue subgenomic replicon into viral particles in highly efficient packaging cells. The subgenomic replicon was constructed by deleting the capsid protein (C) gene from the dengue viral genome and optimizing the signal peptide sequence of pre-membrane protein (prM) to facilitate the formation of viral particles. Packaging cells were developed for inducible expression of a bi-protein Cpr, where the protein pr is the "pr" segment of viral protein prM that holds the protein C on the endoplasmic reticulum (ER). When the replicon was introduced into the packaging cells, protein C was released from the bi-protein Cpr by a replicon-encoded viral protease. Coordinate expression of viral structural proteins by the replicon and packaging cells led to the incorporation of the replicon into viral particle to produce PIVs. Animal tests showed that the dengue PIV vaccine was highly immunogenic and the immune response protected mice challenged with a hundred-fold LD50 inoculation of dengue virus. The method described here has the potential to be applied to vaccine development for other flaviviruses.
KW - Dengue vaccine
KW - Immunogenicity
KW - Packaging cell
KW - Pseudoinfectious virus
KW - Replicon
UR - http://www.scopus.com/inward/record.url?scp=84922254899&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84922254899&partnerID=8YFLogxK
U2 - 10.1016/j.vaccine.2014.03.091
DO - 10.1016/j.vaccine.2014.03.091
M3 - Article
C2 - 24797700
AN - SCOPUS:84922254899
VL - 32
SP - 3854
EP - 3860
JO - Vaccine
JF - Vaccine
SN - 0264-410X
IS - 30
ER -