Calreticulin as a hydrophilic chimeric molecular adjuvant enhances IgG responses to the spike protein of severe acute respiratory syndrome coronavirus
Identifieur interne : 000601 ( Istex/Corpus ); précédent : 000600; suivant : 000602Calreticulin as a hydrophilic chimeric molecular adjuvant enhances IgG responses to the spike protein of severe acute respiratory syndrome coronavirus
Auteurs : Xiang Qiu ; Chao Hong ; Yue Li ; Wanrong Bao ; Xiao-Ming GaoSource :
- Microbiology and Immunology [ 0385-5600 ] ; 2012-08.
English descriptors
- Teeft :
- Adjuvant, Adjuvanticity, Amino acids, Antibody responses, Antigen domain, Bacillus anthracis, Blackwell publishing asia, Calreticulin, Cell epitopes, Cell responses, Coli, Coli cells, Coronavirus, Dendritic cells, Dilution buffer, Encoding, Endoplasmic reticulum, Epitope, Fusion protein, Heat shock proteins, Immunogenicity, Inclusion bodies, Iptg induction, Medical sciences, Molecular adjuvant, Murine, Nucleocapsid protein, Nude mice, Optical density, Present study, Previous investigators, Recombinant, Recombinant calreticulin, Recombinant protein, Recombinant proteins, Renaturation steps, Respiratory coronavirus, Respiratory syndrome, Respiratory syndrome coronavirus spike protein, Resultant products, Sars patients, Serum samples, Soochow university, Spike, Spike protein, Syndrome, Synergistic effect, Synergistic roles, Systemic lupus erythematosus, Triplicate wells, Tumor antigen, Uorescence protein, Vaccine, Vaccine encoding, Viral proteins, Wang, Zhao.
Abstract
Fragment 450–650 of the spike (S) protein (S450–650) of severe acute respiratory syndrome‐associated coronavirus (SARS‐CoV) contains epitopes capable of being recognized by convalescent sera of SARS patients. Vaccination of mice with recombinant S450–650 (rS450–650) can induce Abs against SARS‐CoV, although the titer is relatively low. In the present study, a fusion protein linking a fragment (residues 39–272) of murine calreticulin (CRT) to S450–650 in a prokaryotic expression system was created. Compared with target antigen alone, the recombinant fusion product (rS450–650‐CRT) has much improved hydrophilicity and immunogenicity. The S450–650‐specific IgG Abs of BALB/c mice subcutaneously immunized with rS450–650‐CRT were in substantially higher titer (approximately fivefold more). Furthermore, the fusion protein, but not rS450–650 alone, was able to elicit S450–650‐specific IgG responses in T cell deficient nude mice. Given that rCRT/39–272 can drive the maturation of bone‐marrow‐derived dendritic cells, directly activate macrophages and B cells, and also elicit helper T cell responses in vivo, we propose that fragment 39–272 of CRT is an effective molecular adjuvant capable of enhancing target Ag‐specific humoral responses in both a T cell‐dependent and independent manner. Fusion protein rS450–650‐CRT is a potential candidate vaccine against SARS‐CoV infection.
Url:
DOI: 10.1111/j.1348-0421.2012.00467.x
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<abstract>Fragment 450–650 of the spike (S) protein (S450–650) of severe acute respiratory syndrome‐associated coronavirus (SARS‐CoV) contains epitopes capable of being recognized by convalescent sera of SARS patients. Vaccination of mice with recombinant S450–650 (rS450–650) can induce Abs against SARS‐CoV, although the titer is relatively low. In the present study, a fusion protein linking a fragment (residues 39–272) of murine calreticulin (CRT) to S450–650 in a prokaryotic expression system was created. Compared with target antigen alone, the recombinant fusion product (rS450–650‐CRT) has much improved hydrophilicity and immunogenicity. The S450–650‐specific IgG Abs of BALB/c mice subcutaneously immunized with rS450–650‐CRT were in substantially higher titer (approximately fivefold more). Furthermore, the fusion protein, but not rS450–650 alone, was able to elicit S450–650‐specific IgG responses in T cell deficient nude mice. Given that rCRT/39–272 can drive the maturation of bone‐marrow‐derived dendritic cells, directly activate macrophages and B cells, and also elicit helper T cell responses in vivo, we propose that fragment 39–272 of CRT is an effective molecular adjuvant capable of enhancing target Ag‐specific humoral responses in both a T cell‐dependent and independent manner. Fusion protein rS450–650‐CRT is a potential candidate vaccine against SARS‐CoV infection.</abstract>
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<p>Fragment 450–650 of the spike (S) protein (S450–650) of severe acute respiratory syndrome‐associated coronavirus (SARS‐CoV) contains epitopes capable of being recognized by convalescent sera of SARS patients. Vaccination of mice with recombinant S450–650 (rS450–650) can induce Abs against SARS‐CoV, although the titer is relatively low. In the present study, a fusion protein linking a fragment (residues 39–272) of murine calreticulin (CRT) to S450–650 in a prokaryotic expression system was created. Compared with target antigen alone, the recombinant fusion product (rS450–650‐CRT) has much improved hydrophilicity and immunogenicity. The S450–650‐specific IgG Abs of BALB/c mice subcutaneously immunized with rS450–650‐CRT were in substantially higher titer (approximately fivefold more). Furthermore, the fusion protein, but not rS450–650 alone, was able to elicit S450–650‐specific IgG responses in T cell deficient nude mice. Given that rCRT/39–272 can drive the maturation of bone‐marrow‐derived dendritic cells, directly activate macrophages and B cells, and also elicit helper T cell responses <hi rend="italic">in vivo</hi>
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<titleGroup><title type="main">Calreticulin as a hydrophilic chimeric molecular adjuvant enhances IgG responses to the spike protein of severe acute respiratory syndrome coronavirus</title>
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<p>Fragment 450–650 of the spike (S) protein (S450–650) of severe acute respiratory syndrome‐associated coronavirus (SARS‐CoV) contains epitopes capable of being recognized by convalescent sera of SARS patients. Vaccination of mice with recombinant S450–650 (rS450–650) can induce Abs against SARS‐CoV, although the titer is relatively low. In the present study, a fusion protein linking a fragment (residues 39–272) of murine calreticulin (CRT) to S450–650 in a prokaryotic expression system was created. Compared with target antigen alone, the recombinant fusion product (rS450–650‐CRT) has much improved hydrophilicity and immunogenicity. The S450–650‐specific IgG Abs of BALB/c mice subcutaneously immunized with rS450–650‐CRT were in substantially higher titer (approximately fivefold more). Furthermore, the fusion protein, but not rS450–650 alone, was able to elicit S450–650‐specific IgG responses in T cell deficient nude mice. Given that rCRT/39–272 can drive the maturation of bone‐marrow‐derived dendritic cells, directly activate macrophages and B cells, and also elicit helper T cell responses <i>in vivo</i>
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<abstract lang="en">Fragment 450–650 of the spike (S) protein (S450–650) of severe acute respiratory syndrome‐associated coronavirus (SARS‐CoV) contains epitopes capable of being recognized by convalescent sera of SARS patients. Vaccination of mice with recombinant S450–650 (rS450–650) can induce Abs against SARS‐CoV, although the titer is relatively low. In the present study, a fusion protein linking a fragment (residues 39–272) of murine calreticulin (CRT) to S450–650 in a prokaryotic expression system was created. Compared with target antigen alone, the recombinant fusion product (rS450–650‐CRT) has much improved hydrophilicity and immunogenicity. The S450–650‐specific IgG Abs of BALB/c mice subcutaneously immunized with rS450–650‐CRT were in substantially higher titer (approximately fivefold more). Furthermore, the fusion protein, but not rS450–650 alone, was able to elicit S450–650‐specific IgG responses in T cell deficient nude mice. Given that rCRT/39–272 can drive the maturation of bone‐marrow‐derived dendritic cells, directly activate macrophages and B cells, and also elicit helper T cell responses in vivo, we propose that fragment 39–272 of CRT is an effective molecular adjuvant capable of enhancing target Ag‐specific humoral responses in both a T cell‐dependent and independent manner. Fusion protein rS450–650‐CRT is a potential candidate vaccine against SARS‐CoV infection.</abstract>
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