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In Vitro Generation of Stem Cell Memory-Like T Cells from Activated T Cells

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T-Cell Receptor Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2111))

Abstract

Adoptive T-cell therapy is an attractive strategy for cancer immunotherapy. The transfer of in vitro expanded tumor-associated antigen (TAA)-specific T cells from patients may effectively fight against the original tumor cells. The chimeric antigen receptor-engineered T (CAR-T) cells are also shown to be a promising therapy for hematologic malignancies. However, one of the limitations of these T-cell-based therapies is a rapid acquisition of tolerant (anergy, deletion, dysfunctional and/or exhausted) phenotypes of T cells during activation in vitro and/or after transfer in vivo. We and others found that stem cell memory T (TSCM) cells are strongly resistant against such tolerance, showing strong expansion and persistence in vivo, and provide long-lasting antitumor effects. Here we describe a protocol for the generation of phenotypically TSCM-like cells (iTSCM cells), which can be induced by simple co-culture of activated T cells with OP9 stroma cells expressing a Notch ligand. We also showed the methods of cancer immunotherapy by using NSG mice.

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References

  1. June CH (2007) Adoptive T cell therapy for cancer in the clinic. J Clin Invest 117(6):1466–1476

    Article  CAS  Google Scholar 

  2. Rosenberg SA, Dudley ME (2009) Adoptive cell therapy for the treatment of patients with metastatic melanoma. Curr Opin Immunol 21(2):233–240

    Article  CAS  Google Scholar 

  3. June CH et al (2018) CAR T cell immunotherapy for human cancer. Science 359(6382):1361–1365

    Article  CAS  Google Scholar 

  4. Dunbar CE et al (2018) Gene therapy comes of age. Science 359(6372):eaan4672

    Article  Google Scholar 

  5. June CH, Sadelain M (2018) Chimeric antigen receptor therapy. N Engl J Med 379(1):64–73

    Article  CAS  Google Scholar 

  6. Gattinoni L et al (2005) Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells. J Clin Invest 115(6):1616–1626

    Article  CAS  Google Scholar 

  7. Levine BL et al (2017) Global manufacturing of CAR T cell therapy. Mol Ther Methods Clin Dev 4:92–101

    Article  CAS  Google Scholar 

  8. Louis CU et al (2011) Antitumor activity and long-term fate of chimeric antigen receptor-positive T cells in patients with neuroblastoma. Blood 118(23):6050–6056

    Article  CAS  Google Scholar 

  9. Hinrichs CS et al (2009) Adoptively transferred effector cells derived from naive rather than central memory CD8+ T cells mediate superior antitumor immunity. Proc Natl Acad Sci U S A 106(41):17469–17474

    Article  CAS  Google Scholar 

  10. Berger C et al (2008) Adoptive transfer of effector CD8+ T cells derived from central memory cells establishes persistent T cell memory in primates. J Clin Invest 118(1):294–305

    Article  CAS  Google Scholar 

  11. Gattinoni L et al (2011) A human memory T cell subset with stem cell-like properties. Nat Med 17(10):1290–U325

    Article  CAS  Google Scholar 

  12. Alvarez-Fernandez C et al (2016) A short CD3/CD28 costimulation combined with IL-21 enhance the generation of human memory stem T cells for adoptive immunotherapy. J Transl Med 14(1):214

    Article  CAS  Google Scholar 

  13. Cieri N et al (2013) IL-7 and IL-15 instruct the generation of human memory stem T cells from naive precursors. Blood 121(4):573–584

    Article  CAS  Google Scholar 

  14. Hurton LV et al (2016) Tethered IL-15 augments antitumor activity and promotes a stem-cell memory subset in tumor-specific T cells. Proc Natl Acad Sci U S A 113(48):E7788–E7797

    Article  CAS  Google Scholar 

  15. Scholz G et al (2016) Modulation of mTOR signalling triggers the formation of stem cell-like memory T cells. EBioMedicine 4:50–61

    Article  Google Scholar 

  16. Sabatino M et al (2016) Generation of clinical-grade CD19-specific CAR-modified CD8+ memory stem cells for the treatment of human B-cell malignancies. Blood 128(4):519–528

    Article  CAS  Google Scholar 

  17. Nikolich-Zugich J (2014) Aging of the T cell compartment in mice and humans: from no naive expectations to foggy memories. J Immunol 193(6):2622–2629

    Article  CAS  Google Scholar 

  18. Kondo T et al (2017) Notch-mediated conversion of activated T cells into stem cell memory-like T cells for adoptive immunotherapy. Nat Commun 8:15338

    Article  CAS  Google Scholar 

  19. Kondo T et al (2018) Generation and application of human induced-stem cell memory T cells for adoptive immunotherapy. Cancer Sci 109(7):2130–2140

    Article  CAS  Google Scholar 

  20. Kubuschok B et al (2002) Use of spontaneous Epstein-Barr virus-lymphoblastoid cell lines genetically modified to express tumor antigen as cancer vaccines: mutated p21 ras oncogene in pancreatic carcinoma as a model. Hum Gene Ther 13(7):815–827

    Article  CAS  Google Scholar 

  21. Hui-Yuen J et al (2011) Establishment of Epstein-Barr virus growth-transformed lymphoblastoid cell lines. J Vis Exp 57:3321

    Google Scholar 

  22. Chosewood LC et al (2009) Biosafety in microbiological and biomedical laboratories, 5th edn. U.S. Dept. of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, National Institutes of Health, Washington, D.C., U.S.

    Google Scholar 

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Correspondence to Akihiko Yoshimura .

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Ando, M., Ikeda, M., Yoshimura, A., Kondo, T. (2020). In Vitro Generation of Stem Cell Memory-Like T Cells from Activated T Cells. In: Liu, C. (eds) T-Cell Receptor Signaling. Methods in Molecular Biology, vol 2111. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0266-9_11

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  • DOI: https://doi.org/10.1007/978-1-0716-0266-9_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0265-2

  • Online ISBN: 978-1-0716-0266-9

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