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A kinetic investigation of interacting, stimulated T cells identifies conditions for rapid functional enhancement, minimal phenotype differentiation, and improved adoptive cell transfer tumor eradication

Zhou, Jing and Bethune, Michael T. and Malkova, Natalia and Sutherland, Alexander M. and Comin-Anduix, Begonya and Su, Yapeng and Baltimore, David and Ribas, Antoni and Heath, James R. (2018) A kinetic investigation of interacting, stimulated T cells identifies conditions for rapid functional enhancement, minimal phenotype differentiation, and improved adoptive cell transfer tumor eradication. PLOS ONE, 13 (1). Art. No. e0191634. ISSN 1932-6203. PMCID PMC5779691. https://resolver.caltech.edu/CaltechAUTHORS:20180129-092440324

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[img] MS Word (S1 Method. Immunohistochemistry) - Supplemental Material
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[img] MS Word (S2 Method. Ensemble cytokine secretion detection) - Supplemental Material
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[img] MS Word (S3 Method. Tumor infiltrating lymphocytes (TIL) characterization) - Supplemental Material
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[img] MS Word (S4 Method. Flow cytometry analysis of mouse samples) - Supplemental Material
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[img] MS Word (S5 Method. RNA-seq analysis) - Supplemental Material
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[img] MS Word (S1 Fig. Phenotype dynamics of antigen specific OT1 CD8+ T cells) - Supplemental Material
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[img] MS Word (S2 Fig. Transcriptome dynamics of OT1 CD8+ T cells) - Supplemental Material
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[img] MS Word (S3 Fig. Enriched transcriptional network of OT1 CD8+ T cells from transcriptome analysis) - Supplemental Material
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[img] MS Word (S4 Fig. Enriched biological processes of OT1 CD8+ T cells from transcriptome analysis as T1 is increased) - Supplemental Material
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[img] MS Word (S5 Fig. Gene dynamics of OT1 CD8+ T cells that are highly correlated with effector-vs-memory regulation) - Supplemental Material
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[img] MS Word (S6 Fig. Enriched transcription factors (A) and biological processes (B) by genes that are regulated in the same way in comparison of effector CD8 T cells versus memory CD8 T cells as T1 increases) - Supplemental Material
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[img] MS Word (S7 Fig. In vivo antitumor efficacy with peptide control vs. selected conditions in Fig 2) - Supplemental Material
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[img] MS Word (S8 Fig. Gross cell morphology of EG.7 tumor 4 days after ACT under various conditions) - Supplemental Material
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[img] MS Word (S9 Fig. The level of proliferation in EG.7 tumor after 4 days after ACT under various conditions) - Supplemental Material
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[img] MS Word (S10 Fig. Quantification of CD137 staining) - Supplemental Material
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[img] MS Word (S11 Fig. Population of OT1+ cells that infiltrate into the tumor site) - Supplemental Material
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[img] MS Word (S12 Fig. Ensemble cytokine secretion during T1 = 16 h for different molecular stimulation (anti-CD3 + anti-CD28, left) and (OT1 tetramer + anti-CD28)) - Supplemental Material
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[img] MS Word (S13 Fig. Enlarged luciferase images displayed in Fig 2C) - Supplemental Material
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[img] MS Word (S14 Fig. Relative number of cells that secrete a particular cytokine for human CD4+ T cells) - Supplemental Material
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[img] MS Word (S15 Fig. Enriched transcriptional network of human CD8+ T cells from transcriptome analysis) - Supplemental Material
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[img] MS Word (S16 Fig. Enriched biological processes of human CD8+ T cells from transcriptome analysis as T1 is increased from 0 hour (NS) to 2 hours (A) and from 2 hours to 4 hours (B)) - Supplemental Material
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[img] MS Word (S17 Fig. Gene dynamics of human CD8+ T cells that are highly correlated with effector-vs-memory regulation) - Supplemental Material
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[img] MS Word (S19 Fig. Correlation of human (A) CD4+ and (B) CD8+ T cells from the same patient as T1 increases from 0 hour (NS) to 4 hours) - Supplemental Material
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[img] MS Word (S20 Fig. Phenotypic evolution and transcriptome dynamics for human CD4+ T cells after various T1 conditioning time) - Supplemental Material
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[img] MS Word (S21 Fig. Cytokine secretion dynamics and calibration curve) - Supplemental Material
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[img] MS Word (S22 Fig. Cell clustering dynamics with strong molecular stimulation (OT1 tetramer + anti-CD28 + PMA + ionomycin)) - Supplemental Material
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[img] MS Word (S23 Fig. The RNA expression level of IL2 receptor (IL2RA) as a function of T1 conditioning time) - Supplemental Material
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[img] MS Word (S2 Table. List of antibody panel for human T cells in single cell barcode chip (SCBC)) - Supplemental Material
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[img] MS Word (S3 Table. List of antibody panel for mouse T cells in SCBC) - Supplemental Material
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Abstract

For adoptive cell transfer (ACT) immunotherapy of tumor-reactive T cells, an effective therapeutic outcome depends upon cell dose, cell expansion in vivo through a minimally differentiated phenotype, long term persistence, and strong cytolytic effector function. An incomplete understanding of the biological coupling between T cell expansion, differentiation, and response to stimulation hinders the co-optimization of these factors. We report on a biophysical investigation of how the short-term kinetics of T cell functional activation, through molecular stimulation and cell-cell interactions, competes with phenotype differentiation. T cells receive molecular stimulation for a few minutes to a few hours in bulk culture. Following this priming period, the cells are then analyzed at the transcriptional level, or isolated as single cells, with continuing molecular stimulation, within microchambers for analysis via 11-plex secreted protein assays. We resolve a rapid feedback mechanism, promoted by T cell—T cell contact interactions, which strongly amplifies T cell functional performance while yielding only minimal phenotype differentiation. When tested in mouse models of ACT, optimally primed T cells lead to complete tumor eradication. A similar kinetic process is identified in CD8^+ and CD4^+ T cells collected from a patient with metastatic melanoma.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1371/journal.pone.0191634DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5779691/PubMed CentralArticle
https://doi.org/10.1371/journal.pone.0191634.s001DOIS1 Method. Immunohistochemistry
https://doi.org/10.1371/journal.pone.0191634.s002DOIS2 Method. Ensemble cytokine secretion detection
https://doi.org/10.1371/journal.pone.0191634.s003DOIS3 Method. Tumor infiltrating lymphocytes (TIL) characterization
https://doi.org/10.1371/journal.pone.0191634.s004DOIS4 Method. Flow cytometry analysis of mouse samples
https://doi.org/10.1371/journal.pone.0191634.s005DOIS5 Method. RNA-seq analysis
https://doi.org/10.1371/journal.pone.0191634.s006DOIS1 Fig. Phenotype dynamics of antigen specific OT1 CD8+ T cells
https://doi.org/10.1371/journal.pone.0191634.s007DOIS2 Fig. Transcriptome dynamics of OT1 CD8+ T cells
https://doi.org/10.1371/journal.pone.0191634.s008DOIS3 Fig. Enriched transcriptional network of OT1 CD8+ T cells from transcriptome analysis
https://doi.org/10.1371/journal.pone.0191634.s009DOIS4 Fig. Enriched biological processes of OT1 CD8+ T cells from transcriptome analysis as T1 is increased
https://doi.org/10.1371/journal.pone.0191634.s010DOIS5 Fig. Gene dynamics of OT1 CD8+ T cells that are highly correlated with effector-vs-memory regulation
https://doi.org/10.1371/journal.pone.0191634.s011DOIS6 Fig. Enriched transcription factors (A) and biological processes (B) by genes that are regulated in the same way in comparison of effector CD8 T cells versus memory CD8 T cells as T1 increases
https://doi.org/10.1371/journal.pone.0191634.s012DOIS7 Fig. In vivo antitumor efficacy with peptide control vs. selected conditions in Fig 2
https://doi.org/10.1371/journal.pone.0191634.s013DOIS8 Fig. Gross cell morphology of EG.7 tumor 4 days after ACT under various conditions
https://doi.org/10.1371/journal.pone.0191634.s014DOIS9 Fig. The level of proliferation in EG.7 tumor after 4 days after ACT under various conditions
https://doi.org/10.1371/journal.pone.0191634.s015DOIS10 Fig. Quantification of CD137 staining
https://doi.org/10.1371/journal.pone.0191634.s016DOIS11 Fig. Population of OT1+ cells that infiltrate into the tumor site
https://doi.org/10.1371/journal.pone.0191634.s017DOIS12 Fig. Ensemble cytokine secretion during T1 = 16 h for different molecular stimulation (anti-CD3 + anti-CD28, left) and (OT1 tetramer + anti-CD28)
https://doi.org/10.1371/journal.pone.0191634.s018DOIS13 Fig. Enlarged luciferase images displayed in Fig 2C
https://doi.org/10.1371/journal.pone.0191634.s019DOIS14 Fig. Relative number of cells that secrete a particular cytokine for human CD4+ T cells
https://doi.org/10.1371/journal.pone.0191634.s020DOIS15 Fig. Enriched transcriptional network of human CD8+ T cells from transcriptome analysis
https://doi.org/10.1371/journal.pone.0191634.s021DOIS16 Fig. Enriched biological processes of human CD8+ T cells from transcriptome analysis as T1 is increased from 0 hour (NS) to 2 hours (A) and from 2 hours to 4 hours (B)
https://doi.org/10.1371/journal.pone.0191634.s022DOIS17 Fig. Gene dynamics of human CD8+ T cells that are highly correlated with effector-vs-memory regulation
https://doi.org/10.1371/journal.pone.0191634.s023DOIS18 Fig. Enriched transcription factors (A) and biological processes (B) by genes that are regulated in the same way in comparison of effector CD8 T cells versus memory CD8+ T cells as T1 increases
https://doi.org/10.1371/journal.pone.0191634.s024DOIS19 Fig. Correlation of human (A) CD4+ and (B) CD8+ T cells from the same patient as T1 increases from 0 hour (NS) to 4 hours
https://doi.org/10.1371/journal.pone.0191634.s025DOIS20 Fig. Phenotypic evolution and transcriptome dynamics for human CD4+ T cells after various T1 conditioning time
https://doi.org/10.1371/journal.pone.0191634.s026DOIS21 Fig. Cytokine secretion dynamics and calibration curve
https://doi.org/10.1371/journal.pone.0191634.s027DOIS22 Fig. Cell clustering dynamics with strong molecular stimulation (OT1 tetramer + anti-CD28 + PMA + ionomycin)
https://doi.org/10.1371/journal.pone.0191634.s028DOIS23 Fig. The RNA expression level of IL2 receptor (IL2RA) as a function of T1 conditioning time
https://doi.org/10.1371/journal.pone.0191634.s029DOIS1 Table. List of antibody panel used for flow cytometry immuno-phenotyping analysis for human patient sample
https://doi.org/10.1371/journal.pone.0191634.s030DOIS2 Table. List of antibody panel for human T cells in single cell barcode chip (SCBC)
https://doi.org/10.1371/journal.pone.0191634.s031DOIS3 Table. List of antibody panel for mouse T cells in SCBC
ORCID:
AuthorORCID
Baltimore, David0000-0001-8723-8190
Heath, James R.0000-0001-5356-4385
Additional Information:© 2018 Zhou et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: October 25, 2017; Accepted: January 9, 2018; Published: January 23, 2018. Data Availability Statement: All RNA-seq data files are publicly available at https://www.ncbi.nlm.nih. gov/Traces/study/?acc=SRP126680, accession: PRJNA422284. Funding: This work was supported by National Cancer Institute, grant #'s: R01 CA170689, 5U54 CA119347, 1U54 CA199090-01, CA-16042, AI-28697; Jean Perkins Foundation; and Stand Up 2 Cancer SU2C-AACR-DT1012. The authors thank Georgi K. Marinov and Barbara Wold for valuable discussions and help on the analysis of the RNA-seq data, Igor Antoshechkin for technical support for performing RNA-seq in Millard and Muriel Jacobs Genetics and Genomics Laboratory at Caltech. We also thank Young Shik Shin and Chao Ma for the help on the initial stage of the experiments; Min Xue and Jun Wang for the help on preparing reagents; Julia B Sun for help on data acquisition, Siwen Hu-Lieskovan and Stephen Mok for suggestions for the mice experiments, and Kellin Haley for technical services for flow cytometry studies for human patient samples, which were performed in the UCLA Jonsson Comprehensive Cancer Center (JCCC) and Center for AIDS Research Flow Cytometry Core Facility.
Funders:
Funding AgencyGrant Number
NIHR01 CA170689-01
NIH5U54 CA119347
NIH1U54 CA199090-01
NIHCA-16042
NIHAI-28697
Jean Perkins FoundationUNSPECIFIED
Stand Up 2 CancerSU2C-AACR-DT1012
National Cancer InstituteUNSPECIFIED
Issue or Number:1
PubMed Central ID:PMC5779691
Record Number:CaltechAUTHORS:20180129-092440324
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180129-092440324
Official Citation:Zhou J, Bethune MT, Malkova N, Sutherland AM, Comin-Anduix B, Su Y, et al. (2018) A kinetic investigation of interacting, stimulated T cells identifies conditions for rapid functional enhancement, minimal phenotype differentiation, and improved adoptive cell transfer tumor eradication. PLoS ONE 13(1): e0191634. https://doi.org/10.1371/journal.pone.0191634
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:84559
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:30 Jan 2018 13:23
Last Modified:17 Dec 2019 18:49

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