Virus-Like Particles of SARS-Like Coronavirus Formed by Membrane Proteins from Different Origins Demonstrate Stimulating Activity in Human Dendritic Cells
Identifieur interne : 003037 ( Main/Merge ); précédent : 003036; suivant : 003038Virus-Like Particles of SARS-Like Coronavirus Formed by Membrane Proteins from Different Origins Demonstrate Stimulating Activity in Human Dendritic Cells
Auteurs : Bingke Bai [République populaire de Chine] ; Qinxue Hu [République populaire de Chine] ; Hui Hu [République populaire de Chine] ; Peng Zhou [République populaire de Chine] ; Zhengli Shi [République populaire de Chine] ; Jin Meng [République populaire de Chine] ; Baojing Lu [République populaire de Chine] ; Yi Huang [République populaire de Chine] ; Panyong Mao [République populaire de Chine] ; Hanzhong Wang [République populaire de Chine]Source :
- PLoS ONE [ 1932-6203 ] ; 2008.
Descripteurs français
- KwdFr :
- Agranulocytes (cytologie), Baculoviridae (métabolisme), Cellules dendritiques (cytologie), Cellules dendritiques (virologie), Cytokines (métabolisme), Différenciation cellulaire, Humains, Interféron gamma (métabolisme), Interleukine-4 (métabolisme), Lymphocytes T CD4+ (métabolisme), Lymphocytes auxiliaires Th1 (cytologie), Phénotype, Plasmides (métabolisme), Protéines membranaires (), Système immunitaire, Virus du SRAS (métabolisme).
- MESH :
- cytologie : Agranulocytes, Cellules dendritiques, Lymphocytes auxiliaires Th1.
- métabolisme : Baculoviridae, Cytokines, Interféron gamma, Interleukine-4, Lymphocytes T CD4+, Plasmides, Virus du SRAS.
- virologie : Cellules dendritiques.
- Différenciation cellulaire, Humains, Phénotype, Protéines membranaires, Système immunitaire.
English descriptors
- KwdEn :
- Baculoviridae (metabolism), CD4-Positive T-Lymphocytes (metabolism), Cell Differentiation, Cytokines (metabolism), Dendritic Cells (cytology), Dendritic Cells (virology), Humans, Immune System, Interferon-gamma (metabolism), Interleukin-4 (metabolism), Leukocytes, Mononuclear (cytology), Membrane Proteins (chemistry), Phenotype, Plasmids (metabolism), SARS Virus (metabolism), Th1 Cells (cytology).
- MESH :
- chemical , chemistry : Membrane Proteins.
- chemical , metabolism : Cytokines, Interferon-gamma, Interleukin-4.
- cytology : Dendritic Cells, Leukocytes, Mononuclear, Th1 Cells.
- metabolism : Baculoviridae, CD4-Positive T-Lymphocytes, Plasmids, SARS Virus.
- virology : Dendritic Cells.
- Cell Differentiation, Humans, Immune System, Phenotype.
Abstract
The pathogenesis of SARS coronavirus (CoV) remains poorly understood. In the current study, two recombinant baculovirus were generated to express the spike (S) protein of SARS-like coronavirus (SL-CoV) isolated from bats (vAcBS) and the envelope (E) and membrane (M) proteins of SARS-CoV, respectively. Co-infection of insect cells with these two recombinant baculoviruses led to self-assembly of virus-like particles (BVLPs) as demonstrated by electron microscopy. Incorporation of S protein of vAcBS (BS) into VLPs was confirmed by western blot and immunogold labeling. Such BVLPs up-regulated the level of CD40, CD80, CD86, CD83, and enhanced the secretion of IL-6, IL-10 and TNF-α in immature dendritic cells (DCs). Immune responses were compared in immature DCs inoculated with BVLPs or with VLPs formed by S, E and M proteins of human SARS-CoV. BVLPs showed a stronger ability to stimulate DCs in terms of cytokine induction as evidenced by 2 to 6 fold higher production of IL-6 and TNF-α. Further study indicated that IFN-γ+ and IL-4+ populations in CD4+ T cells increased upon co-cultivation with DCs pre-exposed with BVLPs or SARS-CoV VLPs. The observed difference in DC-stimulating activity between BVLPs and SARS CoV VLPs was very likely due to the S protein. In agreement, SL-CoV S DNA vaccine evoked a more vigorous antibody response and a stronger T cell response than SARS-CoV S DNA in mice. Our data have demonstrated for the first time that SL-CoV VLPs formed by membrane proteins of different origins, one from SL-CoV isolated from bats (BS) and the other two from human SARS-CoV (E and M), activated immature DCs and enhanced the expression of co-stimulatory molecules and the secretion of cytokines. Finding in this study may provide important information for vaccine development as well as for understanding the pathogenesis of SARS-like CoV.
Url:
DOI: 10.1371/journal.pone.0002685
PubMed: 18628832
PubMed Central: 2441860
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type="région">Hubei</region></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff3"><addr-line>Department of Virology, Institute of Infectious Disease, Beijing, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Virology, Institute of Infectious Disease, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff4"><addr-line>Chinese People's Liberation Army Postgraduate Medical School, Beijing, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese People's Liberation Army Postgraduate Medical School, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Hu, Qinxue" sort="Hu, Qinxue" uniqKey="Hu Q" first="Qinxue" last="Hu">Qinxue Hu</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan</wicri:regionArea><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation></author><author><name sortKey="Hu, Hui" sort="Hu, Hui" uniqKey="Hu H" first="Hui" last="Hu">Hui Hu</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan</wicri:regionArea><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation></author><author><name sortKey="Zhou, Peng" sort="Zhou, Peng" uniqKey="Zhou P" first="Peng" last="Zhou">Peng Zhou</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan</wicri:regionArea><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff2"><addr-line>Graduate School of the Chinese Academy of Sciences, Beijing, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Graduate School of the Chinese Academy of Sciences, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Shi, Zhengli" sort="Shi, Zhengli" uniqKey="Shi Z" first="Zhengli" last="Shi">Zhengli Shi</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan</wicri:regionArea><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation></author><author><name sortKey="Meng, Jin" sort="Meng, Jin" uniqKey="Meng J" first="Jin" last="Meng">Jin Meng</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan</wicri:regionArea><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff2"><addr-line>Graduate School of the Chinese Academy of Sciences, Beijing, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Graduate School of the Chinese Academy of Sciences, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Lu, Baojing" sort="Lu, Baojing" uniqKey="Lu B" first="Baojing" last="Lu">Baojing Lu</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan</wicri:regionArea><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation><affiliation wicri:level="3"><nlm:aff id="aff2"><addr-line>Graduate School of the Chinese Academy of Sciences, Beijing, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Graduate School of the Chinese Academy of Sciences, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Huang, Yi" sort="Huang, Yi" uniqKey="Huang Y" first="Yi" last="Huang">Yi Huang</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan</wicri:regionArea><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation></author><author><name sortKey="Mao, Panyong" sort="Mao, Panyong" uniqKey="Mao P" first="Panyong" last="Mao">Panyong Mao</name><affiliation wicri:level="3"><nlm:aff id="aff3"><addr-line>Department of Virology, Institute of Infectious Disease, Beijing, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Virology, Institute of Infectious Disease, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Wang, Hanzhong" sort="Wang, Hanzhong" uniqKey="Wang H" first="Hanzhong" last="Wang">Hanzhong Wang</name><affiliation wicri:level="3"><nlm:aff id="aff1"><addr-line>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People's Republic of China</addr-line></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan</wicri:regionArea><placeName><settlement type="city">Wuhan</settlement><region type="région">Hubei</region></placeName></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2008">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Baculoviridae (metabolism)</term><term>CD4-Positive T-Lymphocytes (metabolism)</term><term>Cell Differentiation</term><term>Cytokines (metabolism)</term><term>Dendritic Cells (cytology)</term><term>Dendritic Cells (virology)</term><term>Humans</term><term>Immune System</term><term>Interferon-gamma (metabolism)</term><term>Interleukin-4 (metabolism)</term><term>Leukocytes, Mononuclear (cytology)</term><term>Membrane Proteins (chemistry)</term><term>Phenotype</term><term>Plasmids (metabolism)</term><term>SARS Virus (metabolism)</term><term>Th1 Cells (cytology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Agranulocytes (cytologie)</term><term>Baculoviridae (métabolisme)</term><term>Cellules dendritiques (cytologie)</term><term>Cellules dendritiques (virologie)</term><term>Cytokines (métabolisme)</term><term>Différenciation cellulaire</term><term>Humains</term><term>Interféron gamma (métabolisme)</term><term>Interleukine-4 (métabolisme)</term><term>Lymphocytes T CD4+ (métabolisme)</term><term>Lymphocytes auxiliaires Th1 (cytologie)</term><term>Phénotype</term><term>Plasmides (métabolisme)</term><term>Protéines membranaires ()</term><term>Système immunitaire</term><term>Virus du SRAS (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Membrane Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Cytokines</term><term>Interferon-gamma</term><term>Interleukin-4</term></keywords><keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Agranulocytes</term><term>Cellules dendritiques</term><term>Lymphocytes auxiliaires Th1</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Dendritic Cells</term><term>Leukocytes, Mononuclear</term><term>Th1 Cells</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Baculoviridae</term><term>CD4-Positive T-Lymphocytes</term><term>Plasmids</term><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Baculoviridae</term><term>Cytokines</term><term>Interféron gamma</term><term>Interleukine-4</term><term>Lymphocytes T CD4+</term><term>Plasmides</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Cellules dendritiques</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Dendritic Cells</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cell Differentiation</term><term>Humans</term><term>Immune System</term><term>Phenotype</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Différenciation cellulaire</term><term>Humains</term><term>Phénotype</term><term>Protéines membranaires</term><term>Système immunitaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The pathogenesis of SARS coronavirus (CoV) remains poorly understood. In the current study, two recombinant baculovirus were generated to express the spike (S) protein of SARS-like coronavirus (SL-CoV) isolated from bats (vAcBS) and the envelope (E) and membrane (M) proteins of SARS-CoV, respectively. Co-infection of insect cells with these two recombinant baculoviruses led to self-assembly of virus-like particles (BVLPs) as demonstrated by electron microscopy. Incorporation of S protein of vAcBS (BS) into VLPs was confirmed by western blot and immunogold labeling. Such BVLPs up-regulated the level of CD40, CD80, CD86, CD83, and enhanced the secretion of IL-6, IL-10 and TNF-α in immature dendritic cells (DCs). Immune responses were compared in immature DCs inoculated with BVLPs or with VLPs formed by S, E and M proteins of human SARS-CoV. BVLPs showed a stronger ability to stimulate DCs in terms of cytokine induction as evidenced by 2 to 6 fold higher production of IL-6 and TNF-α. Further study indicated that IFN-γ+ and IL-4+ populations in CD4+ T cells increased upon co-cultivation with DCs pre-exposed with BVLPs or SARS-CoV VLPs. The observed difference in DC-stimulating activity between BVLPs and SARS CoV VLPs was very likely due to the S protein. In agreement, SL-CoV S DNA vaccine evoked a more vigorous antibody response and a stronger T cell response than SARS-CoV S DNA in mice. Our data have demonstrated for the first time that SL-CoV VLPs formed by membrane proteins of different origins, one from SL-CoV isolated from bats (BS) and the other two from human SARS-CoV (E and M), activated immature DCs and enhanced the expression of co-stimulatory molecules and the secretion of cytokines. 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