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Enhancing lentiviral transduction to generate melanoma-specific human T cells for cancer immunotherapy.

Abstract

Introduction of a tumor antigen-specific T cell receptor (TCR) into patient-derived lymphocytes has already exhibited promising results for the treatment of melanoma and other malignancies in clinical trials. However, insufficient or unsuccessful ex vivo manufacturing of engineered T cells due to low expansion and/or transduction rate can still be observed in some patients. Thus, we isolated human CD8+ T cells from healthy donors and equipped them with a gp100-specific TCR using a lentiviral construct in combination with a novel chemical lentiviral transduction enhancer (Lentiboost) to increase the rate of transduced cells. Following experiments to determine the ideal multiplicity of infection (MOI) and to analyze the efficacy of the transduction enhancer using a GFP-encoding lentivirus, we analyzed in the next step the transduction rate, cell count, and functionality of gp100 TCR-transduced T cells, i.e. antigen-specific cytokine secretion and lytic capacity. In order to increase the number of transduced cells, antigen-specific stimulation was performed, either once for 1 week (1st activation) or twice for another week (2nd activation). In general, each cycle of antigen-specific stimulation resulted in expansion of TCR-positive cells, while no further significant increase of transduced cells was observed after 2nd activation. Cytokine production pattern of transduced cells after antigen encounter, however, revealed significant antigen-specific secretion of TNF and IFNγ after the 1st as well as the 2nd activation. Furthermore, TCR T cells, either activated once or twice, showed significant cytotoxicity towards antigen-positive tumor cells. Taken together, these results show that it is feasible to transduce human T cells using a lentiviral construct in combination with this novel lentiviral transduction enhancer, which shows potential in the growing field of cancer immunotherapy.

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  • Authors+Show Affiliations

    ,

    Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Universitätsklinikum Erlangen, Department of Dermatology, Erlangen, Germany; Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Division of Genetics, Department of Biology, Erlangen, Germany.

    ,

    Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Universitätsklinikum Erlangen, Department of Dermatology, Erlangen, Germany.

    ,

    Sirion Biotech GmbH, Planegg-Martinsried, Germany.

    ,

    Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Universitätsklinikum Erlangen, Department of Dermatology, Erlangen, Germany.

    ,

    Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Universitätsklinikum Erlangen, Department of Dermatology, Erlangen, Germany.

    Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Universitätsklinikum Erlangen, Department of Dermatology, Erlangen, Germany. Electronic address: ugur.uslu@uk-erlangen.de.

    Source

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    31207210

    Citation

    Simon, Bianca, et al. "Enhancing Lentiviral Transduction to Generate Melanoma-specific Human T Cells for Cancer Immunotherapy." Journal of Immunological Methods, 2019.
    Simon B, Harrer DC, Thirion C, et al. Enhancing lentiviral transduction to generate melanoma-specific human T cells for cancer immunotherapy. J Immunol Methods. 2019.
    Simon, B., Harrer, D. C., Thirion, C., Schuler-Thurner, B., Schuler, G., & Uslu, U. (2019). Enhancing lentiviral transduction to generate melanoma-specific human T cells for cancer immunotherapy. Journal of Immunological Methods, doi:10.1016/j.jim.2019.06.015.
    Simon B, et al. Enhancing Lentiviral Transduction to Generate Melanoma-specific Human T Cells for Cancer Immunotherapy. J Immunol Methods. 2019 Jun 14; PubMed PMID: 31207210.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Enhancing lentiviral transduction to generate melanoma-specific human T cells for cancer immunotherapy. AU - Simon,Bianca, AU - Harrer,Dennis C, AU - Thirion,Christian, AU - Schuler-Thurner,Beatrice, AU - Schuler,Gerold, AU - Uslu,Ugur, Y1 - 2019/06/14/ PY - 2019/04/26/received PY - 2019/06/06/revised PY - 2019/06/12/accepted PY - 2019/6/18/pubmed PY - 2019/6/18/medline PY - 2019/6/18/entrez KW - Adoptive T cell therapy KW - Lentiboost KW - Lentivirus KW - T cell receptor KW - Tumor antigen KW - gp100 JF - Journal of immunological methods JO - J. Immunol. Methods N2 - Introduction of a tumor antigen-specific T cell receptor (TCR) into patient-derived lymphocytes has already exhibited promising results for the treatment of melanoma and other malignancies in clinical trials. However, insufficient or unsuccessful ex vivo manufacturing of engineered T cells due to low expansion and/or transduction rate can still be observed in some patients. Thus, we isolated human CD8+ T cells from healthy donors and equipped them with a gp100-specific TCR using a lentiviral construct in combination with a novel chemical lentiviral transduction enhancer (Lentiboost) to increase the rate of transduced cells. Following experiments to determine the ideal multiplicity of infection (MOI) and to analyze the efficacy of the transduction enhancer using a GFP-encoding lentivirus, we analyzed in the next step the transduction rate, cell count, and functionality of gp100 TCR-transduced T cells, i.e. antigen-specific cytokine secretion and lytic capacity. In order to increase the number of transduced cells, antigen-specific stimulation was performed, either once for 1 week (1st activation) or twice for another week (2nd activation). In general, each cycle of antigen-specific stimulation resulted in expansion of TCR-positive cells, while no further significant increase of transduced cells was observed after 2nd activation. Cytokine production pattern of transduced cells after antigen encounter, however, revealed significant antigen-specific secretion of TNF and IFNγ after the 1st as well as the 2nd activation. Furthermore, TCR T cells, either activated once or twice, showed significant cytotoxicity towards antigen-positive tumor cells. Taken together, these results show that it is feasible to transduce human T cells using a lentiviral construct in combination with this novel lentiviral transduction enhancer, which shows potential in the growing field of cancer immunotherapy. SN - 1872-7905 UR - https://www.unboundmedicine.com/medline/citation/31207210/Enhancing_lentiviral_transduction_to_generate_melanoma-specific_human_T_cells_for_cancer_immunotherapy L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-1759(19)30172-3 DB - PRIME DP - Unbound Medicine ER -