國家衛生研究院 NHRI:Item 3990099045/9407

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Title:	The C-terminus of the core beta-ladder domain in Japanese encephalitis virus nonstructural protein 1 is flexible for accommodation of heterologous epitope fusion
Other Titles:	The C-terminus of the core β-ladder domain in Japanese encephalitis virus nonstructural protein 1 is flexible for accommodation of heterologous epitope fusion
Authors:	Yen, LC;Liao, JT;Lee, HJ;Chou, WY;Chen, CW;Lin, YL;Liao, CL
Contributors:	Division of Infectious Diseases
Abstract:	NS1 is the only nonstructural protein that enters the lumen of the endoplasmic reticulum (ER), where NS1 is glycosylated, forms a dimer, and is subsequently secreted during flavivirus replication as dimers or hexamers, which appear to be highly immunogenic to the infected host, as protective immunity can be elicited against homologous flavivirus infections. Here, by using a trans-complementation assay, we identified the C-terminal end of NS1 derived from the Japanese encephalitis virus (JEV), which was more flexible than other regions in terms of housing foreign epitopes without a significant impact on virus replication. This mapped flexible region is located in the conserved tip of the core beta-ladder domain of the multimeric NS1 structure, and is also known to contain certain linear epitopes, readily triggering specific antibody responses from the host. Despite becoming attenuated, recombinant JEV with insertion of a neutralizing epitope derived from Enterovirus-71 (EV71) into the C-terminal end of NS1 could not only be normally released from the infected cells, but also induced dual protective immunity for the host to counteract the lethal challenge of either JEV or EV71 in neonatal mice. These results indicated that the secreted multimeric NS1 of flaviviruses may serve as a natural protein carrier to render epitopes of interest more immunogenic in the C-terminus of the core beta-ladder domain. IMPORTANCE: The positive-sense RNA genome of mosquito-borne flaviviruses appears to be flexible in terms of accommodating extra insertions of short heterologous antigens into their virus genes. Herein, we illustrate that the newly identified C-terminal of the core beta-ladder domain in NS1 could be readily inserted into entities such as EV-71 epitopes, and the resulting NS1-epitope fusion proteins appeared to maintain normal virus replication, secretion ability, and multimeric formation from infected cells. Nonetheless, such an insertion attenuated the recombinant JEV in mice, despite having retained the brain replication ability observed in wild-type JEV. Mother dams immunized with recombinant JEV expressing EV71 epitope-NS1 fused proteins elicited neutralizing antibodies that protected the newborn mice against lethal EV71 challenge. Together, our results implied a potential application of using JEV NS1 as a viral carrier protein to express a heterologous epitope to stimulate dual/multiple protective immunity concurrently against several pathogens.
Date:	2016-02
Relation:	Journal of Virology. 2016 Feb;90(3):1178-1189.
Link to:	http://dx.doi.org/10.1128/jvi.02057-15
JIF/Ranking 2023:	http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=0022-538X&DestApp=IC2JCR
Cited Times(WOS):	https://www.webofscience.com/wos/woscc/full-record/WOS:000369150800003
Cited Times(Scopus):	http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84958064157
Appears in Collections:	[Ching-Len Liao] Periodical Articles

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