Large libraries of single-chain trimer peptide-MHCs enable antigen-specific CD8+ T cell discovery and analysis.
Large libraries of single-chain trimer peptide-MHCs enable antigen-specific CD8+ T cell discovery and analysis.
复制标题
DOI：
10.1038/s42003-023-04899-8
复制
发表时间：
2023-05-16
影响因子：
5.9
通讯作者：
Heath, James R. R.
中科院分区：
文献类型：
--
作者：
Chour, William;Choi, Jongchan;Xie, Jingyi;Chaffee, Mary E. E.;Schmitt, Thomas M. M.;Finton, Kathryn;DeLucia, Diana C. C.;Xu, Alexander M. M.;Su, Yapeng;Chen, Daniel G. G.;Zhang, Rongyu;Yuan, Dan;Hong, Sunga;Ng, Alphonsus H. C.;Butler, Jonah Z. Z.;Edmark, Rick A. A.;Jones, Lesley C. C.;Murray, Kim M. K. M.;Peng, Songming;Li, Guideng;Strong, Roland K. K.;Lee, John K. K.;Goldman, Jason D. D.;Greenberg, Philip D. D.;Heath, James R. R.
抗原特异性CD8+ T细胞克隆型的发现和表征通常涉及肽- mhc四聚体的劳动密集型合成和构建。我们将单链三聚体（SCT）技术应用于pMHC库生成的高通量平台，表明可以跨多个I类HLA等位基因快速制备数百个。我们使用这个平台来探索肽和SCT模板突变对蛋白质表达量、热稳定性和功能的影响。SCT文库是鉴定T细胞识别常见的病毒表位的有效工具。然后，我们构建SCT文库，从COVID-19参与者和健康供体中捕获SARS-CoV-2特异性CD8+ T细胞。这些表位的免疫原性是通过使用SCT文库捕获的克隆tcr对T细胞进行功能分析来验证的。这些技术应该能够在多种情况下快速分析基于肽的T细胞反应，包括自身免疫、癌症或传染病。高通量平台能够生产单链三聚体，用于任何肽和I类HLA等位基因的配对，用于快速发现表位和捕获抗原特异性CD8 + T细胞进行功能表征。
The discovery and characterization of antigen-specific CD8+ T cell clonotypes typically involves the labor-intensive synthesis and construction of peptide-MHC tetramers. We adapt single-chain trimer (SCT) technologies into a high throughput platform for pMHC library generation, showing that hundreds can be rapidly prepared across multiple Class I HLA alleles. We use this platform to explore the impact of peptide and SCT template mutations on protein expression yield, thermal stability, and functionality. SCT libraries were an efficient tool for identifying T cells recognizing commonly reported viral epitopes. We then construct SCT libraries to capture SARS-CoV-2 specific CD8+ T cells from COVID-19 participants and healthy donors. The immunogenicity of these epitopes is validated by functional assays of T cells with cloned TCRs captured using SCT libraries. These technologies should enable the rapid analyses of peptide-based T cell responses across several contexts, including autoimmunity, cancer, or infectious disease. A high-throughput platform enables the production of single-chain trimers for any pairing of peptide and Class I HLA allele for rapid discovery of epitopes and capture of antigen-specific CD8 + T cells for functional characterization.
登录
查看更多内容
DOI：
10.1038/mtm.2014.27
发表时间：
2014
期刊：
Molecular therapy. Methods & clinical development
影响因子：
--
作者：
通讯作者：
--
影响因子：
120.1
作者：
Foy, Susan P. P.;Jacoby, Kyle;Bota, Daniela A. A.;Hunter, Theresa;Pan, Zheng;Stawiski, Eric;Ma, Yan;Lu, William;Peng, Songming;Wang, Clifford L. L.;Yuen, Benjamin;Dalmas, Olivier;Heeringa, Katharine;Sennino, Barbara;Conroy, Andy;Bethune, Michael T. T.;Mende, Ines;White, William;Kukreja, Monica;Gunturu, Swetha;Humphrey, Emily;Hussaini, Adeel;An, Duo;Litterman, Adam J. J.;Quach, Boi Bryant;Ng, Alphonsus H. C.;Lu, Yue;Smith, Chad;Campbell, Katie M. M.;Anaya, Daniel;Skrdlant, Lindsey;Huang, Eva Yi-Hsuan;Mendoza, Ventura;Mathur, Jyoti;Dengler, Luke;Purandare, Bhamini;Moot, Robert;Yi, Michael C. C.;Funke, Roel;Sibley, Alison;Stallings-Schmitt, Todd;Oh, David Y. Y.;Chmielowski, Bartosz;Abedi, Mehrdad;Yuan, Yuan;Sosman, Jeffrey A. A.;Lee, Sylvia M. M.;Schoenfeld, Adam J. J.;Baltimore, David;Heath, James R. R.;Franzusoff, Alex;Ribas, Antoni;Rao, Arati V. V.;Mandl, Stefanie J. J.
通讯作者：
Mandl, Stefanie J. J.
影响因子：
32.4
作者：
Ferretti AP;Kula T;Wang Y;Nguyen DMV;Weinheimer A;Dunlap GS;Xu Q;Nabilsi N;Perullo CR;Cristofaro AW;Whitton HJ;Virbasius A;Olivier KJ Jr;Buckner LR;Alistar AT;Whitman ED;Bertino SA;Chattopadhyay S;MacBeath G
通讯作者：
MacBeath G
影响因子：
3.9
作者：
Dudley, ME;Wunderlich, JR;Rosenberg, SA
通讯作者：
Rosenberg, SA
影响因子：
11.2
作者：
Hebeisen, Michael;Schmidt, Julien;Rufer, Nathalie
通讯作者：
Rufer, Nathalie