Rep. 4, 4166 .42.Cohen, C.J., Li, Y.F., El-Gamil, M., Robbins, P.F., Rosenberg, S.A., and Morgan, R.A. (2007). Enhanced antitumor activity of T cells engineeredto express T-cell receptors with a second disulﬁde bond. Cancer Res.67, 3898–3903 . 43.Kuball, J., Dossett, M.L., Wolﬂ, M., Ho, W.Y., Voss, R.-H., Fowler, C., and Greenberg, P.D. (2007). Facilitating matched pairing and expres- sion of TCR chains introduced into human T cells. Blood 109, 2331– 2338 . 44.Wickham, H. (2009). Ggplot2: Elegant Graphics for Data Analysis (Springer) . Cell Reports Medicine 1, 100141, November 17, 2020 9Reportll OPEN ACCESS STAR+METHODS KEY RESOURCES TABLE REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies CD107a-BV786 BD Biosciences Cat # 563869; RRID:AB_2738458 CD137-BV650 BD Biosciences Cat # 564092; RRID:AB_2738586 CD138-PE BD Biosciences Cat # 552026; RRID:AB_394323 CD14-FITC BioLegend Cat # 301804; RRID:AB_400086 CD152 (CTLA-4)-PE Cy5 BD Biosciences Cat # 555854; RRID:AB_396177 CD19-APC-Cy7 BD Biosciences Cat # 557791; RRID:AB_396873 CD19-BUV496 BD Biosciences Cat # 564655; RRID:AB_2744311 CD1c-BV421 BD Biosciences Cat # 565050; RRID:AB_2744319 CD20-BUV805 BD Biosciences Cat # 612905; RRID:AB_2870192 CD22-BV510 BioLegend Cat # 302526; RRID:AB_2721532 CD24-BV650 BD Biosciences Cat # 563720; RRID:AB_2632388 CD25-BV605 BD Biosciences Cat # 562660; RRID:AB_2744343CD25-PerCPCy5.5 BD Biosciences Cat # 560503; RRID:AB_1727453 CD267-APC BioLegend Cat # 311912; RRID:AB_2565423 CD269-PE-CF594 BioLegend Cat # 357512; RRID:AB_2566531 CD26-APC BioLegend Cat # 302710; RRID:AB_10916120 CD26-PECF594 BD Biosciences Cat # 565158; RRID:AB_2739085 CD27-BV711 BD Biosciences Cat # 356430; RRID:AB_2650751 CD27-BV786 BD Biosciences Cat # 563327; RRID:AB_2744353 CD3-BV421 BD Biosciences Cat # 563797; RRID:AB_2744383 CD38-BV711 BioLegend Cat # 303528; RRID:AB_2563811CD3-FITC BioLegend Cat # 300306; RRID:AB_314042 CD45RA-Alexa Fluor 700 BD Biosciences Cat # 560673; RRID:AB_1727496 CD45RO-BV605 BD Biosciences Cat # 562791; RRID:AB_2744411 CD4-BUV496 BD Biosciences Cat # 564652; RRID:AB_2744422 CD56-FITC BioLegend Cat # 362546; RRID:AB_2565964 CD62L-FITC BD Biosciences Cat # 555543; RRID:AB_395927 CD69-BV786 BD Biosciences Cat # 563834; RRID:AB_2738441 CD69-PE Cy7 BD Biosciences Cat # 557745; RRID:AB_396851 CD79a-PE-Cy7 BioLegend Cat # 333510; RRID:AB_2564595 CD8-BUV805 BD Biosciences Cat # 564912; RRID:AB_2744465 Foxp3-PE Cy7 Life Technologies Cat # 25-4776-42; RRID:AB_10804638Gamma-9-APC BioLegend Cat # 331310; RRID:AB_2057504 HLA-DR-PE-Cy5 BioLegend Cat # 307608; RRID:AB_314686 ICOS-BUV395 BD Biosciences Cat # 564777; RRID:AB_2738946 Ig kappa light chain-Alexa Fluor 700 BD Biosciences Cat # 561319; RRID:AB_10611867 Ig lamda light chain-PerCPCy5.5 BioLegend Cat # 316618; RRID:AB_2561512 IgD-BV605 BD Biosciences Cat # 563313; RRID:AB_2738134 IgM-BUV396 BD Biosciences Cat # 563903; RRID:AB_2721269 LIVE/DEAD Fixable Near-IR Dead Cell Stain Kit ThermoFisher Cat # L10119 PD-1-BV510 BD Biosciences Cat # 563076; RRID:AB_2737990PD-1-PE-CF594 Invitrogen Cat # 61-2799-42; RRID:AB_2574598 TCF7-PE BD Biosciences Cat # 564217; RRID:AB_2687845 (Continued on next page ) e1Cell Reports Medicine 1, 100141, November 17, 2020Reportll OPEN ACCESS RESOURCE AVAILABILITY Lead Contact Further information and requests for resources and reagents should be directed and will be fulﬁlled by the Lead Contact, Asaf Poran (asaf.poran@biontech.us ). Materials Availability All unique/stable reagents generated in this study are available from the Lead Contact with a completed Materials Transfer Agreement.Continued REAGENT or RESOURCE SOURCE IDENTIFIER Biological Samples Patient PBMCs from NT-001 trial Ott et al., 2020 N/A Healthy Donor PBMCs Precision for Medicine Cat # 93000-10M Healthy Donor PBMCs StemExpress Cat # LE010F Healthy Donor PBMCs AllCells https://www.allcells.com/tissue- products/leukopak/ Chemicals, Peptides, and Recombinant Proteins CEF viral peptide pool JPT Cat # PM-CEF-EBenzonase Millipore Sigma Cat # 70746 Assay peptides Ott et al., 2020 N/A MHC class I tetramers Ott et al., 2020 N/A Critical Commercial Assays Pan T cell Isolation Kit, human Miltenyi Biotec Cat # 130-096-535RNeasy Plus Micro Kit QIAGEN Cat # 74034 RNeasy Plus Mini Kit QIAGEN Cat # 74134 Qubit RNA HS Assay Kit ThermoFisher Cat # Q32855 KAPA Library Quantiﬁcation Kit Roche Cat # KK4824 MiSeq Reagent Kit v2 Illumina Cat # MS-102-2003 Fixation/Permeabilization Solution Kit BD Biosciences Cat # 554714 Foxp3/Transcription Factor Staining Buffer Set eBioscience Cat # 00-5523-00 AEC substrate-chromogen BD Biosciences Cat # 551951 IFN gamma Human ELISPOT Kit Invitrogen Cat # 88-7386-88 CD3 MicroBeads, human Miltenyi Biotec Cat # 130-050-101Experimental Models: Cell Lines Jurkat, Clone E6-1 ATCC Cat # ATCC-TIB-152 A375 ATCC Cat # ATCC-CRL-1619 293FT Cell Line Thermo Fisher Cat # R70007 Software and Algorithms MiXCR (version 3.0.12) Bolotin et al., 2015 N/AFlowJo (version 10) FlowJo Software (for Windows) Version 10. Becton, Dickinson and Company; 2019.https://www.ﬂowjo.com/ GraphPad Prism (version 7.01) GraphPad Software, La Jolla California, USA https://www.graphpad.com R (version 3.5.1) CRAN https://www.r-project.org/ DescTools (R package) Signorell et al., 2020 N/A Philentropy (R package) Drost et al., 2018 N/A gplots (R package) Warnes et al., 2015 N/A Stats (R package) R Core Team 2018 N/A Survival (R package) Therneau et al., 2000 N/A Survminer (R package) Kassambara et al., 2019 N/A Cell Reports Medicine 1, 100141, November 17, 2020 e2Reportll OPEN ACCESS Data and Code Availability Patient TCR sequencing information will be provided for patients consistent with their institutional informed consent. The code supporting the ﬁndings of this study is available upon request from the Lead Contact.