Comparison of t cell activities mediated by human TCRs and CARs that use the same recognition domains

Daniel T. Harris, Marlies V. Hager, Sheena N. Smith, Qi Cai, Jennifer D. Stone, Philipp Kruger, Melissa Lever, Omer Dushek, Thomas M. Schmitt, Philip D. Greenberg, David M Kranz

Research output: Contribution to journalArticle

Abstract

Adoptive T cell therapies have achieved significant clinical responses, especially in hematopoietic cancers. Two types of receptor systems have been used to redirect the activity of T cells, normal heterodimeric TCRs or synthetic chimeric Ag receptors (CARs). TCRs recognize peptide-HLA complexes whereas CARs typically use an Ab-derived single-chain fragments variable that recognizes cancer-associated cell-surface Ags. Although both receptors mediate diverse effector functions, a quantitative comparison of the sensitivity and signaling capacity of TCRs and CARs has been limited due to their differences in affinities and ligands. In this study we describe their direct comparison by using TCRs that could be formatted either as conventional ab heterodimers, or as single-chain fragments variable constructs linked to CD3z and CD28 signaling domains or to CD3z alone. Two high-affinity TCRs (K D values of ∼50 and 250 nM) against MART1/HLA-A2 or WT1/HLA-A2 were used, allowing MART1 or WT1 peptide titrations to easily assess the impact of Ag density. Although CARs were expressed at higher surface levels than TCRs, they were 10–100-fold less sensitive, even in the absence of the CD8 coreceptor. Mathematical modeling demonstrated that lower CAR sensitivity could be attributed to less efficient signaling kinetics. Furthermore, reduced cytokine secretion observed at high Ag density for both TCRs and CARs suggested a role for negative regulators in both systems. Interestingly, at high Ag density, CARs also mediated greater maximal release of some cytokines, such as IL-2 and IL-6. These results have implications for the next-generation design of receptors used in adoptive T cell therapies.

Original languageEnglish (US)
Pages (from-to)1088-1100
Number of pages13
JournalJournal of Immunology
Volume200
Issue number3
DOIs
StatePublished - Feb 1 2018

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Human Activities
HLA-A2 Antigen
Single-Chain Antibodies
Cell- and Tissue-Based Therapy
T-Lymphocytes
Cytokines
Peptides
Interleukin-2
Interleukin-6
Neoplasms
Ligands

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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Comparison of t cell activities mediated by human TCRs and CARs that use the same recognition domains. / Harris, Daniel T.; Hager, Marlies V.; Smith, Sheena N.; Cai, Qi; Stone, Jennifer D.; Kruger, Philipp; Lever, Melissa; Dushek, Omer; Schmitt, Thomas M.; Greenberg, Philip D.; Kranz, David M.

In: Journal of Immunology, Vol. 200, No. 3, 01.02.2018, p. 1088-1100.

Research output: Contribution to journalArticle

Harris, DT, Hager, MV, Smith, SN, Cai, Q, Stone, JD, Kruger, P, Lever, M, Dushek, O, Schmitt, TM, Greenberg, PD & Kranz, DM 2018, 'Comparison of t cell activities mediated by human TCRs and CARs that use the same recognition domains' Journal of Immunology, vol. 200, no. 3, pp. 1088-1100. https://doi.org/10.4049/jimmunol.1700236
Harris, Daniel T. ; Hager, Marlies V. ; Smith, Sheena N. ; Cai, Qi ; Stone, Jennifer D. ; Kruger, Philipp ; Lever, Melissa ; Dushek, Omer ; Schmitt, Thomas M. ; Greenberg, Philip D. ; Kranz, David M. / Comparison of t cell activities mediated by human TCRs and CARs that use the same recognition domains. In: Journal of Immunology. 2018 ; Vol. 200, No. 3. pp. 1088-1100.
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AU - Smith, Sheena N.

AU - Cai, Qi

AU - Stone, Jennifer D.

AU - Kruger, Philipp

AU - Lever, Melissa

AU - Dushek, Omer

AU - Schmitt, Thomas M.

AU - Greenberg, Philip D.

AU - Kranz, David M

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AB - Adoptive T cell therapies have achieved significant clinical responses, especially in hematopoietic cancers. Two types of receptor systems have been used to redirect the activity of T cells, normal heterodimeric TCRs or synthetic chimeric Ag receptors (CARs). TCRs recognize peptide-HLA complexes whereas CARs typically use an Ab-derived single-chain fragments variable that recognizes cancer-associated cell-surface Ags. Although both receptors mediate diverse effector functions, a quantitative comparison of the sensitivity and signaling capacity of TCRs and CARs has been limited due to their differences in affinities and ligands. In this study we describe their direct comparison by using TCRs that could be formatted either as conventional ab heterodimers, or as single-chain fragments variable constructs linked to CD3z and CD28 signaling domains or to CD3z alone. Two high-affinity TCRs (K D values of ∼50 and 250 nM) against MART1/HLA-A2 or WT1/HLA-A2 were used, allowing MART1 or WT1 peptide titrations to easily assess the impact of Ag density. Although CARs were expressed at higher surface levels than TCRs, they were 10–100-fold less sensitive, even in the absence of the CD8 coreceptor. Mathematical modeling demonstrated that lower CAR sensitivity could be attributed to less efficient signaling kinetics. Furthermore, reduced cytokine secretion observed at high Ag density for both TCRs and CARs suggested a role for negative regulators in both systems. Interestingly, at high Ag density, CARs also mediated greater maximal release of some cytokines, such as IL-2 and IL-6. These results have implications for the next-generation design of receptors used in adoptive T cell therapies.

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