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T cell antigen discovery via trogocytosis

Li, Guideng and Bethune, Michael T. and Wong, Stephanie and Joglekar, Alok V. and Leonard, Michael T. and Wang, Jessica K. and Kim, Jocelyn T. and Cheng, Donghui and Peng, Songming and Zaretsky, Jesse M. and Su, Yapeng and Luo, Yicheng and Heath, James R. and Ribas, Antoni and Witte, Owen N. and Baltimore, David (2019) T cell antigen discovery via trogocytosis. Nature Methods, 16 (2). pp. 183-190. ISSN 1548-7091. PMCID PMC6719556. https://resolver.caltech.edu/CaltechAUTHORS:20181130-094559783

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[img] Image (JPEG) (Supplementary Figure 1: Establishment of Jurkat cells expressing F5 TCR or 1G4 TCR and K562 cells expressing single-chain trimer (SCT) of HLA-A2/MART126-35(A27L) or HLA-A2/NYESO1157-165(C165V)) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 2: Target cell trogocytosis occurs in an antigen-specific manner) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 3: Trogocytosis can be tracked by multiple protein transfer) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 4: Trogocytosis occurs among various TCR and pMHC allele pairs) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 5: Antigen-specific TCR transfer occurs from donor cells to acceptor cells regardless of cell identity) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 7: Target cell trogocytosis is enhanced by coexpression of CD8) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 10: Comparison of trogocytosis capacity of NeoTCR-CD8-Jurkat cells with MART1-K562, USP7mut-K562 or USP7wt-K562 cells) - Supplemental Material
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Abstract

T cell receptor (TCR) ligand discovery is essential for understanding and manipulating immune responses to tumors. We developed a cell-based selection platform for TCR ligand discovery that exploits a membrane transfer phenomenon called trogocytosis. We discovered that T cell membrane proteins are transferred specifically to target cells that present cognate peptide–major histocompatibility complex (MHC) molecules. Co-incubation of T cells expressing an orphan TCR with target cells collectively presenting a library of peptide–MHCs led to specific labeling of cognate target cells, enabling isolation of these target cells and sequencing of the cognate TCR ligand. We validated this method for two clinically employed TCRs and further used the platform to identify the cognate neoepitope for a subject-derived neoantigen-specific TCR. Thus, target cell trogocytosis is a robust tool for TCR ligand discovery that will be useful for studying basic tumor immunology and identifying new targets for immunotherapy.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1038/s41592-018-0305-7DOIArticle
https://rdcu.be/biV9SPublisherFree ReadCube access
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719556PubMed CentralArticle
ORCID:
AuthorORCID
Li, Guideng0000-0003-0840-7262
Joglekar, Alok V.0000-0001-7554-7447
Leonard, Michael T.0000-0001-9084-2647
Wang, Jessica K.0000-0003-1421-4969
Kim, Jocelyn T.0000-0001-8723-8190
Peng, Songming0000-0002-2742-6584
Luo, Yicheng0000-0003-3704-2389
Heath, James R.0000-0001-5356-4385
Witte, Owen N.0000-0003-4461-4533
Baltimore, David0000-0001-8723-8190
Additional Information:© 2019 Springer Nature Publishing AG. Received 29 June 2018; Accepted 13 December 2018; Published 28 January 2019. Data availability: The original NGS DNA-seq data have been deposited in the Sequence Read Archive under accession numbers SRR8217181, SRR8217182 and SRR8217183. The data that support the findings of this study are available from the corresponding author upon request. Source data for Figs. 2, 4 and 5 are available online. We thank I. Antoshechkin (Millard and Muriel Jacobs Genetics and Genomics Laboratory, Caltech) for deep DNA sequencing, and D. Perez, J. Tijerina and R.A. Diamond (Flow Cytometry Facility, Caltech) for cell sorting. This work was supported by the Prostate Cancer Foundation Challenge Award 15CHAL02 to D.B., O.N.W., L.Y. and M.T.B., and the National Cancer Institute (grant 1U54 CA199090-01 to J.R.H.). M.T.B. is the recipient of a Jane Coffin Childs Postdoctoral Fellowship. A.R. was supported by National Institutes of Health (NIH) grant R35 CA197633. G.L. was supported by the Parker Institute for Cancer Immunotherapy. J.T.K. was supported by NIH/National Center for Advancing Translational Science UCLA CTSI grant KL2TR001882. Author Contributions: M.T.B. conceived of the approach. G.L. and M.T.B. designed research. G.L., M.T.B., S.W., A.V.J., M.T.L., J.K.W., J.T.K., Y.S., Y.L. and D.C. performed experiments. S.P., J.M.Z., A.R. and J.R.H. provided critical reagents and analyzed results. O.N.W. analyzed results. G.L., M.T.B. and D.B. analyzed results and wrote the paper. Competing interests: M.T.B., G.L., J.T.K., S.W. and D.B. are co-inventors on a patent application concerning the described technology, which is licensed to PACT Pharma, Inc. J.R.H. and A.R. are directors and consultants of PACT; D.B. is a consultant of PACT and head of their scientific advisory board; M.T.B. and S.P. are employees of PACT; J.M.Z. is a consultant of PACT; and each of the foregoing individuals has equity interests in PACT.
Funders:
Funding AgencyGrant Number
Prostate Cancer Foundation15CHAL02
NIH1U54 CA199090-01
Jane Coffin Childs Memorial Fund for Medical ResearchUNSPECIFIED
NIHR35 CA197633
Parker InstituteUNSPECIFIED
NIHKL2TR001882
Subject Keywords:Adaptive immunity; Antigen processing and presentation; Immunological techniques
Issue or Number:2
PubMed Central ID:PMC6719556
Record Number:CaltechAUTHORS:20181130-094559783
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20181130-094559783
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:91355
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:29 Jan 2019 16:55
Last Modified:09 Mar 2020 13:19

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