CoChat
Sign Up
Cited by 1,815CAR-T cell therapy researchT-cell and B-cell ImmunologyImmunotherapy and Immune Responses

4-1BB costimulation ameliorates T cell exhaustion induced by tonic signaling of chimeric antigen receptors

Open access
Long, Adrienne H, Haso, Waleed M, Shern, Jack F, Wanhainen, Kelsey M, Murgai, Meera, Ingaramo, Maria, Smith, Jillian P, Walker, Alec J +7 more·2015·Nature Medicine
DOI 10.1038/nm.3838PMC PMC4458184
Open this paper in CoChat — free account

Abstract

Chimeric antigen receptors (CARs) targeting CD19 have mediated dramatic antitumor responses in hematologic malignancies, but tumor regression has rarely occurred using CARs targeting other antigens. It remains unknown whether the impressive effects of CD19 CARs relate to greater susceptibility of hematologic malignancies to CAR therapies, or superior functionality of the CD19 CAR itself. We show that tonic CAR CD3-ζ phosphorylation, triggered by antigen-independent clustering of CAR single-chain variable fragments, can induce early exhaustion of CAR T cells that limits antitumor efficacy. Such activation is present to varying degrees in all CARs studied, except the highly effective CD19 CAR. We further determine that CD28 costimulation augments, whereas 4-1BB costimulation reduces, exhaustion induced by persistent CAR signaling. Our results provide biological explanations for the antitumor effects of CD19 CARs and for the observations that CD19 CAR T cells incorporating the 4-1BB costimulatory domain are more persistent than those incorporating CD28 in clinical trials.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/nm.3838

Cites

8 of 8 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 56

  1. [1]Lee DW The Future Is Now: Chimeric Antigen Receptors as New Targeted Therapies for Childhood Cancer Clinical Cancer Research 2012 18 10 2780 2790 22589486 PMC4119811
  2. [2]Sadelain M Brentjens R Rivière I The Basic Principles of Chimeric Antigen Receptor Design Cancer Discovery 2013 3 4 388 398 23550147 PMC3667586
  3. [5]Kochenderfer JN B-cell depletion and remissions of malignancy along with cytokine-associated toxicity in a clinical trial of anti-CD19 chimeric-antigen-receptor-transduced T cells Blood 2012 119 12 2709 2720 22160384 PMC3327450
  4. [6]Porter DL Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia New England Journal of Medicine 2011 365 8 725 733 21830940 PMC3387277
  5. [7]Grupp SA Chimeric Antigen Receptor-Modified T Cells for Acute Lymphoid Leukemia New England Journal of Medicine 2013 368 16 1509 1518 23527958 PMC4058440
  6. [8]Savoldo B CD28 costimulation improves expansion and persistence of chimeric antigen receptor-modified T cells in lymphoma patients The Journal of Clinical Investigation 2011 121 5 1822 1826 21540550 PMC3083795
  7. [9]Brentjens RJ CD19-Targeted T Cells Rapidly Induce Molecular Remissions in Adults with Chemotherapy-Refractory Acute Lymphoblastic Leukemia Science Translational Medicine 2013 5 177 177ra138 177ra138 PMC3742551 23515080
  8. [10]Davila ML Efficacy and Toxicity Management of 19-28z CAR T Cell Therapy in B Cell Acute Lymphoblastic Leukemia Science Translational Medicine 2014 6 224 224ra225 PMC4684949 24553386
  9. [11]Kershaw MH A phase I study on adoptive immunotherapy using gene-modified T cells for ovarian cancer Clinical Cancer Research 2006 12 20 6106 6115 17062687 PMC2154351
  10. [12]Park JR Adoptive transfer of chimeric antigen receptor re-directed cytolytic T lymphocyte clones in patients with neuroblastoma Molecular therapy 2007 15 4 825 833 17299405
  11. [13]Pule MA Virus-specific T cells engineered to coexpress tumor-specific receptors: persistence and antitumor activity in individuals with neuroblastoma Nature medicine 2008 14 11 1264 1270 PMC2749734 18978797
  12. [14]Louis CU Antitumor activity and long-term fate of chimeric antigen receptor-positive T cells in patients with neuroblastoma Blood 2011 118 23 6050 6056 21984804 PMC3234664
  13. [15]Till BG CD20-specific adoptive immunotherapy for lymphoma using a chimeric antigen receptor with both CD28 and 4-1BB domains: pilot clinical trial results Blood 2012 119 17 3940 3950 22308288 PMC3350361
  14. [16]Lamers CHJ Treatment of Metastatic Renal Cell Carcinoma With CAIX CAR-engineered T cells: Clinical Evaluation and Management of On-target Toxicity Mol Ther 2013 21 4 904 912 23423337 PMC5189272
  15. [17]Robbins PF Cutting edge: persistence of transferred lymphocyte clonotypes correlates with cancer regression in patients receiving cell transfer therapy The Journal of Immunology 2004 173 12 7125 7130 15585832 PMC2175171
  16. [18]Kowolik CM CD28 Costimulation Provided through a CD19-Specific Chimeric Antigen Receptor Enhances In vivo Persistence and Antitumor Efficacy of Adoptively Transferred T Cells Cancer Research 2006 66 22 10995 11004 17108138
  17. [19]Milone MC Chimeric receptors containing CD137 signal transduction domains mediate enhanced survival of T cells and increased antileukemic efficacy in vivo Molecular Therapy 2009 17 8 1453 1464 19384291 PMC2805264
  18. [20]Haso W Anti-CD22-chimeric antigen receptors targeting B-cell precursor acute lymphoblastic leukemia Blood 2013 121 7 1165 1174 23243285 PMC3575759
  19. [22]Sakuishi K Targeting Tim-3 and PD-1 pathways to reverse T cell exhaustion and restore anti-tumor immunity The Journal of experimental medicine 2010 207 10 2187 2194 20819927 PMC2947065
  20. [23]Baitsch L Exhaustion of tumor-specific CD8+ T cells in metastases from melanoma patients The Journal of clinical investigation 2011 121 6 2350 21555851 PMC3104769
  21. [24]Zhou Q Coexpression of Tim-3 and PD-1 identifies a CD8+ T-cell exhaustion phenotype in mice with disseminated acute myelogenous leukemia Blood 2011 117 17 4501 4510 21385853 PMC3099570
  22. [25]Woo S-R Immune inhibitory molecules LAG-3 and PD-1 synergistically regulate T-cell function to promote tumoral immune escape Cancer research 2012 72 4 917 927 22186141 PMC3288154
  23. [26]Topalian SL Safety, activity, and immune correlates of anti-PD-1 antibody in cancer New England Journal of Medicine 2012 366 26 2443 2454 22658127 PMC3544539
  24. [27]Virgin HW Wherry EJ Ahmed R Redefining chronic viral infection Cell 2009 138 1 30 50 19596234
  25. [29]Rossig C Targeting of GD2-positive tumor cells by human T lymphocytes engineered to express chimeric T-cell receptor genes International journal of cancer 2001 94 2 228 236 11668503
  26. [30]Pulè MA A chimeric T cell antigen receptor that augments cytokine release and supports clonal expansion of primary human T cells Molecular Therapy 2005 12 5 933 941 15979412
  27. [31]Hudecek M The non-signaling extracellular spacer domain of chimeric antigen receptors is decisive for in vivo antitumor activity Cancer immunology research 2014 canimm. 0127.2014 PMC4692801 25212991
  28. [32]Kochenderfer JN Construction and Pre-clinical Evaluation of an Anti-CD19 Chimeric Antigen Receptor Journal of immunotherapy (Hagerstown, Md: 1997) 2009 32 7 689 PMC2747302 19561539
  29. [33]Shin H A Role for the Transcriptional Repressor Blimp-1 in CD8+ T Cell Exhaustion during Chronic Viral Infection Immunity 2009 31 2 309 320 19664943 PMC2747257
  30. [34]Doering TA Network Analysis Reveals Centrally Connected Genes and Pathways Involved in CD8< sup>+</sup> T Cell Exhaustion versus Memory Immunity 2012 37 6 1130 1144 23159438 PMC3749234
  31. [35]Paley MA Progenitor and terminal subsets of CD8+ T cells cooperate to contain chronic viral infection Science 2012 338 6111 1220 1225 23197535 PMC3653769
  32. [36]Friend LD A dose-dependent requirement for the proline motif of CD28 in cellular and humoral immunity revealed by a targeted knockin mutant The Journal of experimental medicine 2006 203 9 2121 2133 16908623 PMC2118406
  33. [37]Dodson LF Targeted knock-in mice expressing mutations of CD28 reveal an essential pathway for costimulation Molecular and cellular biology 2009 29 13 3710 3721 19398586 PMC2698768
  34. [38]Zhao Y A herceptin-based chimeric antigen receptor with modified signaling domains leads to enhanced survival of transduced T lymphocytes and antitumor activity The Journal of Immunology 2009 183 9 5563 5574 19843940 PMC6292203
  35. [39]Nieba L Honegger A Krebber C Plückthun A Disrupting the hydrophobic patches at the antibody variable/constant domain interface: improved in vivo folding and physical characterization of an engineered scFv fragment Protein Engineering 1997 10 4 435 444 9194169
  36. [40]Dolezal O Single-chain Fv multimers of the anti-neuraminidase antibody NC10: the residue at position 15 in the VL domain of the scFv-0 (VL− VH) molecule is primarily responsible for formation of a tetramer-trimer equilibrium Protein engineering 2003 16 1 47 56 12646692
  37. [41]Whitlow M Multivalent Fvs: characterization of single-chain Fv oligomers and preparation of a bispecific Fv Protein Engineering 1994 7 8 1017 1026 7809028
  38. [43]Crawford A Molecular and Transcriptional Basis of CD4+ T Cell Dysfunction during Chronic Infection Immunity 40 2 289 302 24530057 PMC3990591
  39. [44]Guest RD The role of extracellular spacer regions in the optimal design of chimeric immune receptors: evaluation of four different scFvs and antigens Journal of Immunotherapy 2005 28 3 203 15838376
  40. [45]James SE Antigen sensitivity of CD22-specific chimeric TCR is modulated by target epitope distance from the cell membrane The Journal of Immunology 2008 180 10 7028 7038 18453625 PMC2585549
  41. [46]Frigault MJ Identification of chimeric antigen receptors that mediate constitutive or inducible proliferation of T cells Cancer immunology research (in the press) PMC4390458 25600436
  42. [47]Agnellini P Impaired NFAT nuclear translocation results in split exhaustion of virus-specific CD8+ T cell functions during chronic viral infection Proceedings of the National Academy of Sciences 2007 104 11 4565 4570 PMC1815473 17360564
  43. [48]Vezys V 4-1BB signaling synergizes with programmed death ligand 1 blockade to augment CD8 T cell responses during chronic viral infection The Journal of Immunology 2011 187 4 1634 1642 21742975 PMC4404506
  44. [49]Wang C 4-1BBL induces TNF receptor-associated factor 1-dependent Bim modulation in human T cells and is a critical component in the costimulation-dependent rescue of functionally impaired HIV-specific CD8 T cells The Journal of Immunology 2007 179 12 8252 8263 18056369
  45. [52]Pearce EL Enhancing CD8 T-cell memory by modulating fatty acid metabolism Nature 2009 460 7251 103 107 19494812 PMC2803086
  46. [53]van der Windt Gerritje JW Mitochondrial Respiratory Capacity Is a Critical Regulator of CD8+ T Cell Memory Development Immunity 2012 36 1 68 78 22206904 PMC3269311
  47. [54]Araki K mTOR regulates memory CD8 T-cell differentiation Nature 2009 460 7251 108 112 19543266 PMC2710807
  48. [55]Roederer M Nozzi JL Nason MC SPICE: Exploration and analysis of post-cytometric complex multivariate datasets Cytometry Part A 2011 79 2 167 174 PMC3072288 21265010
  49. [56]Morgan RA Recognition of glioma stem cells by genetically modified T cells targeting EGFRvIII and development of adoptive cell therapy for glioma Human gene therapy 2012 23 10 1043 1053 22780919 PMC3472555
  50. [57]Brochet X Lefranc M-P Giudicelli V IMGT/V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis Nucleic Acids Research 2008 36 suppl 2 W503 W508 18503082 PMC2447746
  51. [58]Sen G Induction of IgG antibodies by an anti-idiotype antibody mimicking disialoganglioside GD2 Journal of Immunotherapy 1998 21 1 75 83 9456440
  52. [59]Jena B Chimeric Antigen Receptor (CAR)-Specific Monoclonal Antibody to Detect CD19-Specific T Cells in Clinical Trials PLoS ONE 2013 8 3 e57838 23469246 PMC3585808
  53. [60]Edelstein A Computer control of microscopes using µManager Current protocols in molecular biology 2010 14.20. 11-14.20. 17 PMC3065365 20890901
  54. [61]Elangovan M Characterization of one-and two-photon excitation fluorescence resonance energy transfer microscopy Methods 2003 29 1 58 73 12543072
  55. [63]Olenych SG Claxton NS Ottenberg GK Davidson MW The fluorescent protein color palette Current Protocols in Cell Biology 2007 18228502
  56. [64]Martin CE IL-7/anti-IL-7 mAb complexes augment cytokine potency in mice through association with IgG-Fc and by competition with IL-7R Blood 2013 121 22 4484 4492 23610371 PMC3668485

Cited by

Sorted by citations

Authors
Long, A. H., Haso, W. M., Shern, J. F., Wanhainen, K. M., Murgai, M., Ingaramo, M., Smith, J. P., Walker, A. J., Kohler, M. E., Venkateshwara, V. R., Kaplan, R. N., Patterson, G. H., Fry, T. J., Orentas, R. J., & Mackall, C. L.
Affiliations
  • Northwestern University (US)
  • National Institutes of Health (US)
Corresponding author
Mackall, Crystal L
ORCID
Venue
Nature Medicine
Publisher
Springer Science and Business Media LLC
Published
2015-05-04
Volume
21
Issue
6
Pages
581-590
ISSN
  • 1078-8956 (ISSN-L)
  • 1546-170X
DOI
10.1038/nm.3838
PMC
PMC4458184
OpenAlex
W1619537825
Cited by
1,815
References
8
Citations by year
2026: 183 · 2025: 158 · 2024: 201 · 2023: 218 · 2022: 211 · 2021: 217 · 2020: 183 · 2019: 133 · 2018: 116 · 2017: 121 · 2016: 55 · 2015: 15 · 2014: 1 · 2012: 1
FWCI
44.70
Type
journal article
Language
en
Open access
Licence
www.springer.com/tdm (tdm)
Locations
  • Nature Medicine (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health
  • National Cancer Institute (ZIABC011565, ZIABC011073, ZIABC011332)
  • National Institute of Biomedical Imaging and Bioengineering (ZIAEB000071)
Topics
  • Health Sciences › Medicine › Oncology › CAR-T cell therapy research
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
  • Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
Keywords
4-1BB costimulation, T cell exhaustion, chimeric antigen receptor, tonic signaling, CD3, CD28 costimulation, CAR T-cell therapy, CD19, antitumor efficacy, hematologic malignancies
MeSH terms
Humans, Interleukin-2, Lymphocyte Activation, Receptors, Antigen, T-Lymphocytes, Immunotherapy, Adoptive, CD28 Antigens, Antigens, CD19, Hematologic Neoplasms, Cell Line, Tumor, Tumor Necrosis Factor Receptor Superfamily, Member 9

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review