Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published January 29, 2019 | Supplemental Material + Accepted Version
Journal Article Open

T cell antigen discovery via trogocytosis

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.

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.

Attached Files

Accepted Version - nihms-1037677.pdf

Supplemental Material - 41592_2018_305_Fig10_ESM.jpg

Supplemental Material - 41592_2018_305_Fig11_ESM.jpg

Supplemental Material - 41592_2018_305_Fig12_ESM.jpg

Supplemental Material - 41592_2018_305_Fig13_ESM.jpg

Supplemental Material - 41592_2018_305_Fig14_ESM.jpg

Supplemental Material - 41592_2018_305_Fig15_ESM.jpg

Supplemental Material - 41592_2018_305_Fig6_ESM.jpg

Supplemental Material - 41592_2018_305_Fig7_ESM.jpg

Supplemental Material - 41592_2018_305_Fig8_ESM.jpg

Supplemental Material - 41592_2018_305_Fig9_ESM.jpg

Supplemental Material - 41592_2018_305_MOESM1_ESM.pdf

Supplemental Material - 41592_2018_305_MOESM2_ESM.pdf

Supplemental Material - 41592_2018_305_MOESM3_ESM.pdf

Supplemental Material - 41592_2018_305_MOESM4_ESM.xls

Supplemental Material - 41592_2018_305_MOESM5_ESM.xls

Supplemental Material - 41592_2018_305_MOESM6_ESM.xlsx

Supplemental Material - 41592_2018_305_MOESM7_ESM.xlsx

Supplemental Material - 41592_2018_305_MOESM8_ESM.xlsx

Files

41592_2018_305_Fig11_ESM.jpg
Files (13.6 MB)
Name Size Download all
md5:fb35294c4fbd1ef999d3139c3e8e043e
134.6 kB Preview Download
md5:b66a42e9fe667189d38a4ed554fcc8a6
90.8 kB Preview Download
md5:3f5d120d16dbc8cf1a97824072dd36f7
54.3 kB Preview Download
md5:5c3be55883c4862d3dffc98d19d7bd70
10.0 kB Download
md5:8c7b2e68fc4049e3970148da2589d3a0
111.7 kB Preview Download
md5:cd87a796689b45a15ad7683fc7594097
6.0 MB Download
md5:e58dce7e0447bedb0bb04cc50842aa0e
10.7 kB Download
md5:07e8813b1ae75f0b7c9109b05026ed74
94.0 kB Preview Download
md5:46eb006824baa35ae602035521989070
117.4 kB Preview Download
md5:5eff69545189d707c6e5d74a405f6c0d
86.6 kB Preview Download
md5:83c07911b9b3d8c3b259aa65ab091745
100.2 kB Preview Download
md5:23d7f07ad16942fc79988e245bf2642f
59.9 kB Preview Download
md5:43260c6c92891088b8d3c6cf21c2c061
74.3 kB Preview Download
md5:1142089242d8f04717669eba9baf1f90
4.1 MB Preview Download
md5:18584ea5068f387e9a105d4782e75d6b
1.3 MB Download
md5:c0b3a8495241978531590737e58f0925
10.4 kB Download
md5:44ca34f7137a4c28a85365d5281be953
50.5 kB Preview Download
md5:442b7e14884bed1509cf8cc267c74e72
113.4 kB Preview Download
md5:8263f3327477026001bf27e0c33e4725
1.2 MB Preview Download

Additional details

Created:
August 22, 2023
Modified:
October 19, 2023