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A Lentiviral Vector Expressing Japanese Encephalitis Virus-like Particles Elicits Broad Neutralizing Antibody Response in Pigs.

de Wispelaere M, Ricklin M, Souque P, Frenkiel MP, Paulous S, Garcìa-Nicolàs O, Summerfield A, Charneau P, Desprès P - PLoS Negl Trop Dis (2015)

Bottom Line: Vaccination of domestic pigs has been suggested as a "one health" strategy to reduce viral disease transmission to humans.Immunization of pigs with the lentiviral vector expressing JEV virus-like particles is particularly efficient to prime antigen-specific humoral immunity and trigger neutralizing antibody responses against JEV genotypes 1, 3, and 5.The titers of neutralizing antibodies elicited by the TRIP/JEV vector are sufficient to confer protection in domestic pigs against different genotypes of JEV and this could be of a great utility in endemic regions where more than one genotype is circulating.

View Article: PubMed Central - PubMed

Affiliation: Interactions Moléculaires Flavivirus-Hôtes, Institut Pasteur, Paris, France.

ABSTRACT

Background: Japanese encephalitis virus (JEV) is the major cause of viral encephalitis in Southeast Asia. Vaccination of domestic pigs has been suggested as a "one health" strategy to reduce viral disease transmission to humans. The efficiency of two lentiviral TRIP/JEV vectors expressing the JEV envelope prM and E glycoproteins at eliciting protective humoral response was assessed in a mouse model and piglets.

Methodology/principal findings: A gene encoding the envelope proteins prM and E from a genotype 3 JEV strain was inserted into a lentiviral TRIP vector. Two lentiviral vectors TRIP/JEV were generated, each expressing the prM signal peptide followed by the prM protein and the E glycoprotein, the latter being expressed either in its native form or lacking its two C-terminal transmembrane domains. In vitro transduction of cells with the TRIP/JEV vector expressing the native prM and E resulted in the efficient secretion of virus-like particles of Japanese encephalitis virus. Immunization of BALB/c mice with TRIP/JEV vectors resulted in the production of IgGs against Japanese encephalitis virus, and the injection of a second dose one month after the prime injection greatly boosted antibody titers. The TRIP/JEV vectors elicited neutralizing antibodies against JEV strains belonging to genotypes 1, 3, and 5. Immunization of piglets with two doses of the lentiviral vector expressing JEV virus-like particles led to high titers of anti-JEV antibodies, that had efficient neutralizing activity regardless of the JEV genotype tested.

Conclusions/significance: Immunization of pigs with the lentiviral vector expressing JEV virus-like particles is particularly efficient to prime antigen-specific humoral immunity and trigger neutralizing antibody responses against JEV genotypes 1, 3, and 5. The titers of neutralizing antibodies elicited by the TRIP/JEV vector are sufficient to confer protection in domestic pigs against different genotypes of JEV and this could be of a great utility in endemic regions where more than one genotype is circulating.

No MeSH data available.


Related in: MedlinePlus

Schematic representation of recombinant lentiviral TRIP/JEV vectors.The organization of JEV genomic RNA is shown in top. The codon-optimized sequence encoding prM and E from JEV G3 strain RP-9 was cloned into a TRIP lentiviral vector under the control of human cytomegalovirus immediate early promoter (PCMVie). The TRIP/JEV.prME vector includes the signal peptide sequence of prM (SP) followed by the entire prM and E gene regions of JEV strain RP-9 of G3. The TRIP/prMEΔTM vector includes the same JEV sequence except that E was deleted from its two transmembrane domains TMD1 and TMD2.
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pntd.0004081.g001: Schematic representation of recombinant lentiviral TRIP/JEV vectors.The organization of JEV genomic RNA is shown in top. The codon-optimized sequence encoding prM and E from JEV G3 strain RP-9 was cloned into a TRIP lentiviral vector under the control of human cytomegalovirus immediate early promoter (PCMVie). The TRIP/JEV.prME vector includes the signal peptide sequence of prM (SP) followed by the entire prM and E gene regions of JEV strain RP-9 of G3. The TRIP/prMEΔTM vector includes the same JEV sequence except that E was deleted from its two transmembrane domains TMD1 and TMD2.

Mentions: We have reported earlier that a single minute dose of a non-replicative lentiviral vector expressing the soluble form of WNV E glycoprotein induced a robust protective humoral response in a mouse model of WNV encephalitis [17, 18]. To assess the potential of lentiviral vectors expressing JEV proteins at eliciting protective humoral response against JEV infection, a mammalian codon-optimized gene encoding prM and E from the JEV strain RP9 of G3 was inserted into the lentivirus TRIP vector (Fig 1). We generated two lentiviral vectors, expressing the prM signal peptide followed by the prM protein and the E glycoprotein, the latter being expressed either in the native form (TRIP/JEV.prME) or lacking its two C-terminal transmembrane domains (TRIP/JEV.prMEΔTM). In these constructs, prM contributes to the folding, stability, and efficient secretion of the glycoprotein E. Lentiviral vectors which expressed JEV proteins were pseudotyped with VSV-G protein of the IND serotype. Non-replicative TRIP/JEV.prME and TRIP/JEV.prMEΔTM particles were produced from HEK-293T cells, achieving titers of about 7 log10 TU.mL-1.


A Lentiviral Vector Expressing Japanese Encephalitis Virus-like Particles Elicits Broad Neutralizing Antibody Response in Pigs.

de Wispelaere M, Ricklin M, Souque P, Frenkiel MP, Paulous S, Garcìa-Nicolàs O, Summerfield A, Charneau P, Desprès P - PLoS Negl Trop Dis (2015)

Schematic representation of recombinant lentiviral TRIP/JEV vectors.The organization of JEV genomic RNA is shown in top. The codon-optimized sequence encoding prM and E from JEV G3 strain RP-9 was cloned into a TRIP lentiviral vector under the control of human cytomegalovirus immediate early promoter (PCMVie). The TRIP/JEV.prME vector includes the signal peptide sequence of prM (SP) followed by the entire prM and E gene regions of JEV strain RP-9 of G3. The TRIP/prMEΔTM vector includes the same JEV sequence except that E was deleted from its two transmembrane domains TMD1 and TMD2.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4593544&req=5

pntd.0004081.g001: Schematic representation of recombinant lentiviral TRIP/JEV vectors.The organization of JEV genomic RNA is shown in top. The codon-optimized sequence encoding prM and E from JEV G3 strain RP-9 was cloned into a TRIP lentiviral vector under the control of human cytomegalovirus immediate early promoter (PCMVie). The TRIP/JEV.prME vector includes the signal peptide sequence of prM (SP) followed by the entire prM and E gene regions of JEV strain RP-9 of G3. The TRIP/prMEΔTM vector includes the same JEV sequence except that E was deleted from its two transmembrane domains TMD1 and TMD2.
Mentions: We have reported earlier that a single minute dose of a non-replicative lentiviral vector expressing the soluble form of WNV E glycoprotein induced a robust protective humoral response in a mouse model of WNV encephalitis [17, 18]. To assess the potential of lentiviral vectors expressing JEV proteins at eliciting protective humoral response against JEV infection, a mammalian codon-optimized gene encoding prM and E from the JEV strain RP9 of G3 was inserted into the lentivirus TRIP vector (Fig 1). We generated two lentiviral vectors, expressing the prM signal peptide followed by the prM protein and the E glycoprotein, the latter being expressed either in the native form (TRIP/JEV.prME) or lacking its two C-terminal transmembrane domains (TRIP/JEV.prMEΔTM). In these constructs, prM contributes to the folding, stability, and efficient secretion of the glycoprotein E. Lentiviral vectors which expressed JEV proteins were pseudotyped with VSV-G protein of the IND serotype. Non-replicative TRIP/JEV.prME and TRIP/JEV.prMEΔTM particles were produced from HEK-293T cells, achieving titers of about 7 log10 TU.mL-1.

Bottom Line: Vaccination of domestic pigs has been suggested as a "one health" strategy to reduce viral disease transmission to humans.Immunization of pigs with the lentiviral vector expressing JEV virus-like particles is particularly efficient to prime antigen-specific humoral immunity and trigger neutralizing antibody responses against JEV genotypes 1, 3, and 5.The titers of neutralizing antibodies elicited by the TRIP/JEV vector are sufficient to confer protection in domestic pigs against different genotypes of JEV and this could be of a great utility in endemic regions where more than one genotype is circulating.

View Article: PubMed Central - PubMed

Affiliation: Interactions Moléculaires Flavivirus-Hôtes, Institut Pasteur, Paris, France.

ABSTRACT

Background: Japanese encephalitis virus (JEV) is the major cause of viral encephalitis in Southeast Asia. Vaccination of domestic pigs has been suggested as a "one health" strategy to reduce viral disease transmission to humans. The efficiency of two lentiviral TRIP/JEV vectors expressing the JEV envelope prM and E glycoproteins at eliciting protective humoral response was assessed in a mouse model and piglets.

Methodology/principal findings: A gene encoding the envelope proteins prM and E from a genotype 3 JEV strain was inserted into a lentiviral TRIP vector. Two lentiviral vectors TRIP/JEV were generated, each expressing the prM signal peptide followed by the prM protein and the E glycoprotein, the latter being expressed either in its native form or lacking its two C-terminal transmembrane domains. In vitro transduction of cells with the TRIP/JEV vector expressing the native prM and E resulted in the efficient secretion of virus-like particles of Japanese encephalitis virus. Immunization of BALB/c mice with TRIP/JEV vectors resulted in the production of IgGs against Japanese encephalitis virus, and the injection of a second dose one month after the prime injection greatly boosted antibody titers. The TRIP/JEV vectors elicited neutralizing antibodies against JEV strains belonging to genotypes 1, 3, and 5. Immunization of piglets with two doses of the lentiviral vector expressing JEV virus-like particles led to high titers of anti-JEV antibodies, that had efficient neutralizing activity regardless of the JEV genotype tested.

Conclusions/significance: Immunization of pigs with the lentiviral vector expressing JEV virus-like particles is particularly efficient to prime antigen-specific humoral immunity and trigger neutralizing antibody responses against JEV genotypes 1, 3, and 5. The titers of neutralizing antibodies elicited by the TRIP/JEV vector are sufficient to confer protection in domestic pigs against different genotypes of JEV and this could be of a great utility in endemic regions where more than one genotype is circulating.

No MeSH data available.


Related in: MedlinePlus