The expression of membrane protein augments the specific responses induced by SARS-CoV nucleocapsid DNA immunization.
Identifieur interne : 003C65 ( Main/Exploration ); précédent : 003C64; suivant : 003C66The expression of membrane protein augments the specific responses induced by SARS-CoV nucleocapsid DNA immunization.
Auteurs : Shu-Qun Shi [République populaire de Chine] ; Jing-Pian Peng ; Yin-Chuan Li ; Chuan Qin ; Guo-Dong Liang ; Li Xu ; Ying Yang ; Jin-Ling Wang ; Quan-Hong SunSource :
- Molecular immunology [ 0161-5890 ] ; 2006.
Descripteurs français
- KwdFr :
- ADN viral (génétique), Animaux, Anticorps antiviraux (immunologie), Arvicolinae (immunologie), Complexe antigène-anticorps (immunologie), Cytokines (métabolisme), Cytométrie en flux, Expression des gènes, Lymphocytes T CD4+ (cytologie), Lymphocytes T CD4+ (métabolisme), Lymphocytes T CD8+ (cytologie), Lymphocytes T CD8+ (métabolisme), Poumon (cytologie), Prolifération cellulaire, Protéines membranaires (génétique), Protéines membranaires (immunologie), Protéines membranaires (métabolisme), Protéines nucléocapside (génétique), Protéines nucléocapside (immunologie), Protéines nucléocapside (isolement et purification), Souris, Souris de lignée BALB C, Vaccins à ADN (immunologie), Virus du SRAS (génétique), Virus du SRAS (immunologie).
- MESH :
- cytologie : Lymphocytes T CD4+, Lymphocytes T CD8+, Poumon.
- génétique : ADN viral, Protéines membranaires, Protéines nucléocapside, Virus du SRAS.
- immunologie : Anticorps antiviraux, Arvicolinae, Complexe antigène-anticorps, Protéines membranaires, Protéines nucléocapside, Vaccins à ADN, Virus du SRAS.
- isolement et purification : Protéines nucléocapside.
- métabolisme : Cytokines, Lymphocytes T CD4+, Lymphocytes T CD8+, Protéines membranaires.
- Animaux, Cytométrie en flux, Expression des gènes, Prolifération cellulaire, Souris, Souris de lignée BALB C.
English descriptors
- KwdEn :
- Animals, Antibodies, Viral (immunology), Antigen-Antibody Complex (immunology), Arvicolinae (immunology), CD4-Positive T-Lymphocytes (cytology), CD4-Positive T-Lymphocytes (metabolism), CD8-Positive T-Lymphocytes (cytology), CD8-Positive T-Lymphocytes (metabolism), Cell Proliferation, Cytokines (metabolism), DNA, Viral (genetics), Flow Cytometry, Gene Expression, Lung (cytology), Membrane Proteins (genetics), Membrane Proteins (immunology), Membrane Proteins (metabolism), Mice, Mice, Inbred BALB C, Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (immunology), Nucleocapsid Proteins (isolation & purification), SARS Virus (genetics), SARS Virus (immunology), Vaccines, DNA (immunology).
- MESH :
- chemical , genetics : DNA, Viral, Membrane Proteins, Nucleocapsid Proteins.
- chemical , immunology : Antibodies, Viral, Antigen-Antibody Complex, Membrane Proteins, Nucleocapsid Proteins, Vaccines, DNA.
- cytology : CD4-Positive T-Lymphocytes, CD8-Positive T-Lymphocytes, Lung.
- genetics : SARS Virus.
- immunology : Arvicolinae, SARS Virus.
- chemical , isolation & purification : Nucleocapsid Proteins.
- metabolism : CD4-Positive T-Lymphocytes, CD8-Positive T-Lymphocytes, Cytokines, Membrane Proteins.
- Animals, Cell Proliferation, Flow Cytometry, Gene Expression, Mice, Mice, Inbred BALB C.
Abstract
Nucleocapsid protein plays a critical role in SARS-CoV pathogenesis, and high-level anti-nucleocapsid antibodies are detected in the patients infected by severe acute respiratory syndrome-associated coronavirus (SARS-CoV). Several studies have shown that there exists an interaction between nucleocapsid (N) and membrane (M) protein. In this paper, we investigate whether the expression of membrane protein can affect the immune responses induced by nucleocapsid DNA immunization. Two recombinant plasmids containing M and N coding sequence were constructed. Moreover, in order to get the antigen for ELISA and in vitro stimulation assay, N protein were expressed and purified from E. coli bacteria. Injection of 20mug of the mixture of pVAX1-M and pVAX1-N into the Balb/c mice could elicit the humoral and cellular responses. The ELISA analysis using the N antigen or inactivated SARS-CoV particles as capture antigen showed that co-injection of SARS-M could enhance N-induced antibody production, especially IgG2a subclass. After lymphocytes were stimulated with 10mug/ml purified N antigen, The CD4+ and CD8+ T cells of N and M plus N group were increased compared with those of control groups, and the M protein could augment the activation of lymphocytes induced by N DNA vaccine. Cytokine ELISA analysis revealed that co-injection of M could enhance the levels of IFN-gamma, IL-2 release induced by N antigen. Further experiments in field mouse also support the claim that membrane protein can augment the N-specific immune responses. Virus challenge test was conducted in BSL3 bio safety laboratory with Brandt's vole SARS-CoV model, and the results indicated that co-immunization of M and N antigens could reduce the mortality and pathological changes in lung from the virus infection.
DOI: 10.1016/j.molimm.2005.11.005
PubMed: 16423399
Affiliations:
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responses induced by SARS-CoV nucleocapsid DNA immunization.</title><author><name sortKey="Shi, Shu Qun" sort="Shi, Shu Qun" uniqKey="Shi S" first="Shu-Qun" last="Shi">Shu-Qun Shi</name><affiliation wicri:level="1"><nlm:affiliation>State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, PR China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Peng, Jing Pian" sort="Peng, Jing Pian" uniqKey="Peng J" first="Jing-Pian" last="Peng">Jing-Pian Peng</name></author><author><name sortKey="Li, Yin Chuan" sort="Li, Yin Chuan" uniqKey="Li Y" first="Yin-Chuan" last="Li">Yin-Chuan Li</name></author><author><name sortKey="Qin, Chuan" sort="Qin, Chuan" uniqKey="Qin C" first="Chuan" last="Qin">Chuan Qin</name></author><author><name sortKey="Liang, Guo Dong" sort="Liang, Guo Dong" uniqKey="Liang G" first="Guo-Dong" last="Liang">Guo-Dong Liang</name></author><author><name sortKey="Xu, Li" sort="Xu, Li" uniqKey="Xu L" first="Li" last="Xu">Li Xu</name></author><author><name sortKey="Yang, Ying" sort="Yang, Ying" uniqKey="Yang Y" first="Ying" last="Yang">Ying Yang</name></author><author><name sortKey="Wang, Jin Ling" sort="Wang, Jin Ling" uniqKey="Wang J" first="Jin-Ling" last="Wang">Jin-Ling Wang</name></author><author><name sortKey="Sun, Quan Hong" sort="Sun, Quan Hong" uniqKey="Sun Q" first="Quan-Hong" last="Sun">Quan-Hong Sun</name></author></analytic><series><title level="j">Molecular immunology</title><idno type="ISSN">0161-5890</idno><imprint><date when="2006" type="published">2006</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Viral (immunology)</term><term>Antigen-Antibody Complex (immunology)</term><term>Arvicolinae (immunology)</term><term>CD4-Positive T-Lymphocytes (cytology)</term><term>CD4-Positive T-Lymphocytes (metabolism)</term><term>CD8-Positive T-Lymphocytes (cytology)</term><term>CD8-Positive T-Lymphocytes (metabolism)</term><term>Cell Proliferation</term><term>Cytokines (metabolism)</term><term>DNA, Viral (genetics)</term><term>Flow Cytometry</term><term>Gene Expression</term><term>Lung (cytology)</term><term>Membrane Proteins (genetics)</term><term>Membrane Proteins (immunology)</term><term>Membrane Proteins (metabolism)</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Nucleocapsid Proteins (genetics)</term><term>Nucleocapsid Proteins (immunology)</term><term>Nucleocapsid Proteins (isolation & purification)</term><term>SARS Virus (genetics)</term><term>SARS Virus (immunology)</term><term>Vaccines, DNA (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN viral (génétique)</term><term>Animaux</term><term>Anticorps antiviraux (immunologie)</term><term>Arvicolinae (immunologie)</term><term>Complexe antigène-anticorps (immunologie)</term><term>Cytokines (métabolisme)</term><term>Cytométrie en flux</term><term>Expression des gènes</term><term>Lymphocytes T CD4+ (cytologie)</term><term>Lymphocytes T CD4+ (métabolisme)</term><term>Lymphocytes T CD8+ (cytologie)</term><term>Lymphocytes T CD8+ (métabolisme)</term><term>Poumon (cytologie)</term><term>Prolifération cellulaire</term><term>Protéines membranaires (génétique)</term><term>Protéines membranaires (immunologie)</term><term>Protéines membranaires (métabolisme)</term><term>Protéines nucléocapside (génétique)</term><term>Protéines nucléocapside (immunologie)</term><term>Protéines nucléocapside (isolement et purification)</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Vaccins à ADN (immunologie)</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Viral</term><term>Membrane Proteins</term><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Viral</term><term>Antigen-Antibody Complex</term><term>Membrane Proteins</term><term>Nucleocapsid Proteins</term><term>Vaccines, DNA</term></keywords><keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Lymphocytes T CD4+</term><term>Lymphocytes T CD8+</term><term>Poumon</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>CD4-Positive T-Lymphocytes</term><term>CD8-Positive T-Lymphocytes</term><term>Lung</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN viral</term><term>Protéines membranaires</term><term>Protéines nucléocapside</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term><term>Arvicolinae</term><term>Complexe antigène-anticorps</term><term>Protéines membranaires</term><term>Protéines nucléocapside</term><term>Vaccins à ADN</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Arvicolinae</term><term>SARS Virus</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Protéines nucléocapside</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>CD4-Positive T-Lymphocytes</term><term>CD8-Positive T-Lymphocytes</term><term>Cytokines</term><term>Membrane Proteins</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Cytokines</term><term>Lymphocytes T CD4+</term><term>Lymphocytes T CD8+</term><term>Protéines membranaires</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Proliferation</term><term>Flow Cytometry</term><term>Gene Expression</term><term>Mice</term><term>Mice, Inbred BALB C</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cytométrie en flux</term><term>Expression des gènes</term><term>Prolifération cellulaire</term><term>Souris</term><term>Souris de lignée BALB C</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Nucleocapsid protein plays a critical role in SARS-CoV pathogenesis, and high-level anti-nucleocapsid antibodies are detected in the patients infected by severe acute respiratory syndrome-associated coronavirus (SARS-CoV). Several studies have shown that there exists an interaction between nucleocapsid (N) and membrane (M) protein. In this paper, we investigate whether the expression of membrane protein can affect the immune responses induced by nucleocapsid DNA immunization. Two recombinant plasmids containing M and N coding sequence were constructed. Moreover, in order to get the antigen for ELISA and in vitro stimulation assay, N protein were expressed and purified from E. coli bacteria. Injection of 20mug of the mixture of pVAX1-M and pVAX1-N into the Balb/c mice could elicit the humoral and cellular responses. The ELISA analysis using the N antigen or inactivated SARS-CoV particles as capture antigen showed that co-injection of SARS-M could enhance N-induced antibody production, especially IgG2a subclass. After lymphocytes were stimulated with 10mug/ml purified N antigen, The CD4+ and CD8+ T cells of N and M plus N group were increased compared with those of control groups, and the M protein could augment the activation of lymphocytes induced by N DNA vaccine. Cytokine ELISA analysis revealed that co-injection of M could enhance the levels of IFN-gamma, IL-2 release induced by N antigen. Further experiments in field mouse also support the claim that membrane protein can augment the N-specific immune responses. Virus challenge test was conducted in BSL3 bio safety laboratory with Brandt's vole SARS-CoV model, and the results indicated that co-immunization of M and N antigens could reduce the mortality and pathological changes in lung from the virus infection.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Li, Yin Chuan" sort="Li, Yin Chuan" uniqKey="Li Y" first="Yin-Chuan" last="Li">Yin-Chuan Li</name><name sortKey="Liang, Guo Dong" sort="Liang, Guo Dong" uniqKey="Liang G" first="Guo-Dong" last="Liang">Guo-Dong Liang</name><name sortKey="Peng, Jing Pian" sort="Peng, Jing Pian" uniqKey="Peng J" first="Jing-Pian" last="Peng">Jing-Pian Peng</name><name sortKey="Qin, Chuan" sort="Qin, Chuan" uniqKey="Qin C" first="Chuan" last="Qin">Chuan Qin</name><name sortKey="Sun, Quan Hong" sort="Sun, Quan Hong" uniqKey="Sun Q" first="Quan-Hong" last="Sun">Quan-Hong Sun</name><name sortKey="Wang, Jin Ling" sort="Wang, Jin Ling" uniqKey="Wang J" first="Jin-Ling" last="Wang">Jin-Ling Wang</name><name sortKey="Xu, Li" sort="Xu, Li" uniqKey="Xu L" first="Li" last="Xu">Li Xu</name><name sortKey="Yang, Ying" sort="Yang, Ying" uniqKey="Yang Y" first="Ying" last="Yang">Ying Yang</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Shi, Shu Qun" sort="Shi, Shu Qun" uniqKey="Shi S" first="Shu-Qun" last="Shi">Shu-Qun Shi</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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