title: Building and optimizing a virus-specific T cell receptor library for targeted immunotherapy in viral infections.
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.
authors.affiliation: Singapore Institute for Clinical Sciences, Agency for Science Technology and Research (A*Star), Singapore.
authors.forename: Nasirah
authors.initials: N
authors.lastname: Banu
authors.affiliation: Singapore Institute for Clinical Sciences, Agency for Science Technology and Research (A*Star), Singapore.
authors.forename: Adeline
authors.initials: A
authors.lastname: Chia
authors.affiliation: Singapore Institute for Clinical Sciences, Agency for Science Technology and Research (A*Star), Singapore.
authors.forename: Zi Zong
authors.initials: ZZ
authors.lastname: Ho
authors.affiliation: Singapore Institute for Clinical Sciences, Agency for Science Technology and Research (A*Star), Singapore.
authors.forename: Alfonso Tan
authors.initials: AT
authors.lastname: Garcia
authors.affiliation: Singapore Institute for Clinical Sciences, Agency for Science Technology and Research (A*Star), Singapore.
authors.forename: Komathi
authors.initials: K
authors.lastname: Paravasivam
authors.affiliation: Departments of Microbiology and Biological Sciences, Immunology Programme, National University of Singapore, Singapore.
authors.forename: Gijsbert M
authors.initials: GM
authors.lastname: Grotenbreg
authors.affiliation: 1] Singapore Institute for Clinical Sciences, Agency for Science Technology and Research (A*Star), Singapore [2] Program of Emerging Viral Diseases, Duke-NUS Graduate Medical School, National University of Singapore, Singapore.
authors.forename: Antonio
authors.initials: A
authors.lastname: Bertoletti
authors.affiliation: 1] Singapore Institute for Clinical Sciences, Agency for Science Technology and Research (A*Star), Singapore [2] Molecular Microbiology and Immunology, Saint Louis University School of Medicine, Saint Louis, Missouri, USA [3] Saint Louis University Liver Center, Saint Louis University School of Medicine, Saint Louis, Missouri, USA.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Gehring
mesh_terms: D002460:Cell Line; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007167:Immunotherapy; D058990:Molecular Targeted Therapy; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D014161:Transduction, Genetic; D014777:Virus Diseases / Q000235:genetics* / Q000276:immunology / Q000628:therapy
chemical_list: D011948:Receptors, Antigen, T-Cell