Building and Optimizing a Virus-specific T Cell Receptor Library for Targeted Immunotherapy in Viral Infections
Identifieur interne : 000A43 ( Main/Exploration ); précédent : 000A42; suivant : 000A44Building and Optimizing a Virus-specific T Cell Receptor Library for Targeted Immunotherapy in Viral Infections
Auteurs : Nasirah Banu [Singapour] ; Adeline Chia [Singapour] ; Zi Zong Ho [Singapour] ; Alfonso Tan Garcia [Singapour] ; Komathi Paravasivam [Singapour] ; Gijsbert M. Grotenbreg [Singapour] ; Antonio Bertoletti [Singapour] ; Adam J. Gehring [Singapour, États-Unis]Source :
- Scientific Reports [ 2045-2322 ] ; 2014.
Abstract
Restoration of antigen-specific T cell immunity has the potential to clear persistent viral infection. T cell receptor (TCR) gene therapy can reconstitute CD8 T cell immunity in chronic patients. We cloned 10 virus-specific TCRs targeting 5 different viruses, causing chronic and acute infection. All 10 TCR genetic constructs were optimized for expression using a P2A sequence, codon optimization and the addition of a non-native disulfide bond. However, maximum TCR expression was only achieved after establishing the optimal orientation of the alpha and beta chains in the expression cassette; 9/10 TCRs favored the beta-P2A-alpha orientation over alpha-P2A-beta. Optimal TCR expression was associated with a significant increase in the frequency of IFN-gamma+ T cells. In addition, activating cells for transduction in the presence of Toll-like receptor ligands further enhanced IFN-gamma production. Thus, we have built a virus-specific TCR library that has potential for therapeutic intervention in chronic viral infection or virus-related cancers.
Url:
DOI: 10.1038/srep04166
PubMed: 24566718
PubMed Central: 3933865
Affiliations:
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Grotenbreg</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>Departments of Microbiology and Biological Sciences, Immunology Programme, National University of Singapore</institution>, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Bertoletti, Antonio" sort="Bertoletti, Antonio" uniqKey="Bertoletti A" first="Antonio" last="Bertoletti">Antonio Bertoletti</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Singapore Institute for Clinical Sciences, Agency for Science Technology and Research (A*Star)</institution>, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea></wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a3"><institution>Program of Emerging Viral Diseases, Duke-NUS Graduate Medical School, National University of Singapore</institution>, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea></wicri:regionArea></affiliation></author><author><name sortKey="Gehring, Adam J" sort="Gehring, Adam J" uniqKey="Gehring A" first="Adam J." last="Gehring">Adam J. Gehring</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Singapore Institute for Clinical Sciences, Agency for Science Technology and Research (A*Star)</institution>, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea></wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Molecular Microbiology and Immunology, Saint Louis University School of Medicine</institution>, Saint Louis, Missouri,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a5"><institution>Saint Louis University Liver Center, Saint Louis University School of Medicine</institution>, Saint Louis, Missouri,<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Restoration of antigen-specific T cell immunity has the potential to clear persistent viral infection. T cell receptor (TCR) gene therapy can reconstitute CD8 T cell immunity in chronic patients. We cloned 10 virus-specific TCRs targeting 5 different viruses, causing chronic and acute infection. All 10 TCR genetic constructs were optimized for expression using a P2A sequence, codon optimization and the addition of a non-native disulfide bond. However, maximum TCR expression was only achieved after establishing the optimal orientation of the alpha and beta chains in the expression cassette; 9/10 TCRs favored the beta-P2A-alpha orientation over alpha-P2A-beta. Optimal TCR expression was associated with a significant increase in the frequency of IFN-gamma+ T cells. In addition, activating cells for transduction in the presence of Toll-like receptor ligands further enhanced IFN-gamma production. Thus, we have built a virus-specific TCR library that has potential for therapeutic intervention in chronic viral infection or virus-related cancers.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Bertoletti, A" uniqKey="Bertoletti A">A. Bertoletti</name></author><author><name sortKey="Ferrari, C" uniqKey="Ferrari C">C. Ferrari</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pagliara, D" uniqKey="Pagliara D">D. Pagliara</name></author><author><name sortKey="Savoldo, B" uniqKey="Savoldo B">B. Savoldo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lau, G K" uniqKey="Lau G">G. K. Lau</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lau, G K K" uniqKey="Lau G">G. K. K. Lau</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Loggi, E" uniqKey="Loggi E">E. 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Lee</name></author><author><name sortKey="Strominger, J L" uniqKey="Strominger J">J. L. Strominger</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Labrecque, N" uniqKey="Labrecque N">N. Labrecque</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Arber, C" uniqKey="Arber C">C. Arber</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Leen, A M" uniqKey="Leen A">A. M. Leen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chang, C X L" uniqKey="Chang C">C. X. L. Chang</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Singapour</li><li>États-Unis</li></country></list><tree><country name="Singapour"><noRegion><name sortKey="Banu, Nasirah" sort="Banu, Nasirah" uniqKey="Banu N" first="Nasirah" last="Banu">Nasirah Banu</name></noRegion><name sortKey="Bertoletti, Antonio" sort="Bertoletti, Antonio" uniqKey="Bertoletti A" first="Antonio" last="Bertoletti">Antonio Bertoletti</name><name sortKey="Bertoletti, Antonio" sort="Bertoletti, Antonio" uniqKey="Bertoletti A" first="Antonio" last="Bertoletti">Antonio Bertoletti</name><name sortKey="Chia, Adeline" sort="Chia, Adeline" uniqKey="Chia A" first="Adeline" last="Chia">Adeline Chia</name><name sortKey="Garcia, Alfonso Tan" sort="Garcia, Alfonso Tan" uniqKey="Garcia A" first="Alfonso Tan" last="Garcia">Alfonso Tan Garcia</name><name sortKey="Gehring, Adam J" sort="Gehring, Adam J" uniqKey="Gehring A" first="Adam J." last="Gehring">Adam J. Gehring</name><name sortKey="Grotenbreg, Gijsbert M" sort="Grotenbreg, Gijsbert M" uniqKey="Grotenbreg G" first="Gijsbert M." last="Grotenbreg">Gijsbert M. Grotenbreg</name><name sortKey="Ho, Zi Zong" sort="Ho, Zi Zong" uniqKey="Ho Z" first="Zi Zong" last="Ho">Zi Zong Ho</name><name sortKey="Paravasivam, Komathi" sort="Paravasivam, Komathi" uniqKey="Paravasivam K" first="Komathi" last="Paravasivam">Komathi Paravasivam</name></country><country name="États-Unis"><noRegion><name sortKey="Gehring, Adam J" sort="Gehring, Adam J" uniqKey="Gehring A" first="Adam J." last="Gehring">Adam J. Gehring</name></noRegion><name sortKey="Gehring, Adam J" sort="Gehring, Adam J" uniqKey="Gehring A" first="Adam J." last="Gehring">Adam J. Gehring</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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