Target antigen density governs the efficacy of anti-CD20-CD28-CD3 ζ chimeric antigen receptor-modified effector CD8+ T cells

Keisuke Watanabe, Seitaro Terakura, Anton C. Martens, Tom Van Meerten, Susumu Uchiyama, Misa Imai, Reona Sakemura, Tatsunori Goto, Ryo Hanajiri, Nobuhiko Imahashi, Kazuyuki Shimada, Akihiro Tomita, Hitoshi Kiyoi, Tetsuya Nishida, Tomoki Naoe, Makoto Murata

Research output: Contribution to journalArticle

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Abstract

The effectiveness of chimeric Ag receptor (CAR)-transduced T (CAR-T) cells has been attributed to supraphysiological signaling through CARs. Second- and later-generation CARs simultaneously transmit costimulatory signals with CD3z signals upon ligation, but may lead to severe adverse effects owing to the recognition of minimal Ag expression outside the target tumor. Currently, the threshold target Ag density for CAR-T cell lysis and further activation, including cytokine production, has not yet been investigated in detail. Therefore, we determined the threshold target Ag density required to induce CAR-T cell responses using novel anti-CD20 CART cells with a CD28 intracellular domain and a CD20-transduced CEM cell model. The newly developed CD20CAR-T cells demonstrated Ag-specific lysis and cytokine secretion, which was a reasonable level as a second-generation CAR. For lytic activity, the threshold Ag density was determined to be ∼200 molecules per target cell, whereas the Ag density required for cytokine production of CAR-T cells was ∼10-fold higher, at a few thousand per target cell. CD20CAR-T cells responded efficiently to CD20-downregulated lymphoma and leukemia targets, including rituximab-or ofatumumab-refractory primary chronic lymphocytic leukemia cells. Despite the potential influence of the structure, localization, and binding affinity of the CAR/Ag, the threshold determined may be used for target Ag selection. An Ag density below the threshold may not result in adverse effects, whereas that above the threshold may be sufficient for practical effectiveness. CD20CAR-T cells also demonstrated significant lytic activity against CD20-downregulated tumor cells and may exhibit effectiveness for CD20-positive lymphoid malignancies.

Original languageEnglish
Pages (from-to)911-920
Number of pages10
JournalJournal of Immunology
Volume194
Issue number3
DOIs
Publication statusPublished - 01-02-2015

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CD3 Antigens
Antigen Receptors
T-Cell Antigen Receptor
T-Lymphocytes
Antigens
Cytokines
Down-Regulation
Neoplasms
B-Cell Chronic Lymphocytic Leukemia
Ligation
Lymphoma
Leukemia

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Watanabe, K., Terakura, S., Martens, A. C., Van Meerten, T., Uchiyama, S., Imai, M., ... Murata, M. (2015). Target antigen density governs the efficacy of anti-CD20-CD28-CD3 ζ chimeric antigen receptor-modified effector CD8+ T cells. Journal of Immunology, 194(3), 911-920. https://doi.org/10.4049/jimmunol.1402346
Watanabe, Keisuke ; Terakura, Seitaro ; Martens, Anton C. ; Van Meerten, Tom ; Uchiyama, Susumu ; Imai, Misa ; Sakemura, Reona ; Goto, Tatsunori ; Hanajiri, Ryo ; Imahashi, Nobuhiko ; Shimada, Kazuyuki ; Tomita, Akihiro ; Kiyoi, Hitoshi ; Nishida, Tetsuya ; Naoe, Tomoki ; Murata, Makoto. / Target antigen density governs the efficacy of anti-CD20-CD28-CD3 ζ chimeric antigen receptor-modified effector CD8+ T cells. In: Journal of Immunology. 2015 ; Vol. 194, No. 3. pp. 911-920.
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Watanabe, K, Terakura, S, Martens, AC, Van Meerten, T, Uchiyama, S, Imai, M, Sakemura, R, Goto, T, Hanajiri, R, Imahashi, N, Shimada, K, Tomita, A, Kiyoi, H, Nishida, T, Naoe, T & Murata, M 2015, 'Target antigen density governs the efficacy of anti-CD20-CD28-CD3 ζ chimeric antigen receptor-modified effector CD8+ T cells', Journal of Immunology, vol. 194, no. 3, pp. 911-920. https://doi.org/10.4049/jimmunol.1402346

Target antigen density governs the efficacy of anti-CD20-CD28-CD3 ζ chimeric antigen receptor-modified effector CD8+ T cells. / Watanabe, Keisuke; Terakura, Seitaro; Martens, Anton C.; Van Meerten, Tom; Uchiyama, Susumu; Imai, Misa; Sakemura, Reona; Goto, Tatsunori; Hanajiri, Ryo; Imahashi, Nobuhiko; Shimada, Kazuyuki; Tomita, Akihiro; Kiyoi, Hitoshi; Nishida, Tetsuya; Naoe, Tomoki; Murata, Makoto.

In: Journal of Immunology, Vol. 194, No. 3, 01.02.2015, p. 911-920.

Research output: Contribution to journalArticle

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AU - Watanabe, Keisuke

AU - Terakura, Seitaro

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AU - Van Meerten, Tom

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AU - Imai, Misa

AU - Sakemura, Reona

AU - Goto, Tatsunori

AU - Hanajiri, Ryo

AU - Imahashi, Nobuhiko

AU - Shimada, Kazuyuki

AU - Tomita, Akihiro

AU - Kiyoi, Hitoshi

AU - Nishida, Tetsuya

AU - Naoe, Tomoki

AU - Murata, Makoto

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N2 - The effectiveness of chimeric Ag receptor (CAR)-transduced T (CAR-T) cells has been attributed to supraphysiological signaling through CARs. Second- and later-generation CARs simultaneously transmit costimulatory signals with CD3z signals upon ligation, but may lead to severe adverse effects owing to the recognition of minimal Ag expression outside the target tumor. Currently, the threshold target Ag density for CAR-T cell lysis and further activation, including cytokine production, has not yet been investigated in detail. Therefore, we determined the threshold target Ag density required to induce CAR-T cell responses using novel anti-CD20 CART cells with a CD28 intracellular domain and a CD20-transduced CEM cell model. The newly developed CD20CAR-T cells demonstrated Ag-specific lysis and cytokine secretion, which was a reasonable level as a second-generation CAR. For lytic activity, the threshold Ag density was determined to be ∼200 molecules per target cell, whereas the Ag density required for cytokine production of CAR-T cells was ∼10-fold higher, at a few thousand per target cell. CD20CAR-T cells responded efficiently to CD20-downregulated lymphoma and leukemia targets, including rituximab-or ofatumumab-refractory primary chronic lymphocytic leukemia cells. Despite the potential influence of the structure, localization, and binding affinity of the CAR/Ag, the threshold determined may be used for target Ag selection. An Ag density below the threshold may not result in adverse effects, whereas that above the threshold may be sufficient for practical effectiveness. CD20CAR-T cells also demonstrated significant lytic activity against CD20-downregulated tumor cells and may exhibit effectiveness for CD20-positive lymphoid malignancies.

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