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In vitro modelling of local gene therapy with IL-15/IL-15Rα and a PD-L1 antagonist in melanoma reveals an interplay between NK cells and CD4(+) T cells

Blockade of the immune checkpoint axis consisting of programmed death-1 (PD-1) and its ligand PD-L1 alleviates the functional inhibition of tumor-infiltrating lymphoid cells yet weakly induces their expansion. Exogenous cytokines could further expand lymphoid cells and thus synergize with αPD-L1 the...

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Autores principales: Awad, Robin Maximilian, De Vlaeminck, Yannick, Meeus, Fien, Ertveldt, Thomas, Zeven, Katty, Ceuppens, Hannelore, Goyvaerts, Cleo, Verdonck, Magali, Salguero, Gustavo, Raes, Geert, Devoogdt, Nick, Breckpot, Karine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10624833/
https://www.ncbi.nlm.nih.gov/pubmed/37923822
http://dx.doi.org/10.1038/s41598-023-45948-w
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author Awad, Robin Maximilian
De Vlaeminck, Yannick
Meeus, Fien
Ertveldt, Thomas
Zeven, Katty
Ceuppens, Hannelore
Goyvaerts, Cleo
Verdonck, Magali
Salguero, Gustavo
Raes, Geert
Devoogdt, Nick
Breckpot, Karine
author_facet Awad, Robin Maximilian
De Vlaeminck, Yannick
Meeus, Fien
Ertveldt, Thomas
Zeven, Katty
Ceuppens, Hannelore
Goyvaerts, Cleo
Verdonck, Magali
Salguero, Gustavo
Raes, Geert
Devoogdt, Nick
Breckpot, Karine
author_sort Awad, Robin Maximilian
collection PubMed
description Blockade of the immune checkpoint axis consisting of programmed death-1 (PD-1) and its ligand PD-L1 alleviates the functional inhibition of tumor-infiltrating lymphoid cells yet weakly induces their expansion. Exogenous cytokines could further expand lymphoid cells and thus synergize with αPD-L1 therapy. However, systemic delivery of most cytokines causes severe toxicity due to unspecific expansion of immune cells in the periphery. Here, we modelled local delivery of cytokines and αPD-L1 therapeutics to immune cell-containing in vitro melanoma tumors. Three-dimensional tumor models consisting of 624-MEL cells were co-cultured with human peripheral blood lymphoid cells (PBLs) in presence of the cytokines IL-2, IL-7, IL-15, IL-21 and IFN-γ. To model local gene therapy, melanoma tumors were modified with lentiviral vectors encoding IL-15 fused to IL-15Rα (IL-15/IL-15Rα) and K2-Fc, a fusion of a human PD-L1 specific single domain antibody to immunoglobulin (Ig)G1 Fc. To evaluate the interplay between PBL fractions, NK cells, CD4(+) T cells or CD8(+) T cells were depleted. Tumor cell killing was followed up using real time imaging and immune cell expansion and activation was evaluated with flow cytometry. Among the tested cytokines, IL-15 was the most potent cytokine in stimulating tumor cell killing and expanding both natural killer (NK) cells and CD8(+) T cells. Gene-based delivery of IL-15/IL-15Rα to tumor cells, shows expansion of NK cells, activation of NK cells, CD4(+) and CD8(+) T cells, and killing of tumor spheroids. Both NK cells and CD8(+) T cells are necessary for tumor cell killing and CD4(+) T-cell activation was reduced without NK cells. Co-delivery of K2-Fc improved tumor cell killing coinciding with increased activation of NK cells, which was independent of bystander T cells. CD4(+) or CD8(+) T cells were not affected by the co-delivery of K2-Fc even though NK-cell activation impacted CD4(+) T-cell activation. This study demonstrates that gene-based delivery of IL-15/IL-15Rα to tumor cells effectively mediates anti-tumor activity and sensitizes the tumor microenvironment for therapy with αPD-L1 therapeutics mainly by impacting NK cells. These findings warrant further investigation of gene-based IL-15 and K2-Fc delivery in vivo.
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spelling pubmed-106248332023-11-05 In vitro modelling of local gene therapy with IL-15/IL-15Rα and a PD-L1 antagonist in melanoma reveals an interplay between NK cells and CD4(+) T cells Awad, Robin Maximilian De Vlaeminck, Yannick Meeus, Fien Ertveldt, Thomas Zeven, Katty Ceuppens, Hannelore Goyvaerts, Cleo Verdonck, Magali Salguero, Gustavo Raes, Geert Devoogdt, Nick Breckpot, Karine Sci Rep Article Blockade of the immune checkpoint axis consisting of programmed death-1 (PD-1) and its ligand PD-L1 alleviates the functional inhibition of tumor-infiltrating lymphoid cells yet weakly induces their expansion. Exogenous cytokines could further expand lymphoid cells and thus synergize with αPD-L1 therapy. However, systemic delivery of most cytokines causes severe toxicity due to unspecific expansion of immune cells in the periphery. Here, we modelled local delivery of cytokines and αPD-L1 therapeutics to immune cell-containing in vitro melanoma tumors. Three-dimensional tumor models consisting of 624-MEL cells were co-cultured with human peripheral blood lymphoid cells (PBLs) in presence of the cytokines IL-2, IL-7, IL-15, IL-21 and IFN-γ. To model local gene therapy, melanoma tumors were modified with lentiviral vectors encoding IL-15 fused to IL-15Rα (IL-15/IL-15Rα) and K2-Fc, a fusion of a human PD-L1 specific single domain antibody to immunoglobulin (Ig)G1 Fc. To evaluate the interplay between PBL fractions, NK cells, CD4(+) T cells or CD8(+) T cells were depleted. Tumor cell killing was followed up using real time imaging and immune cell expansion and activation was evaluated with flow cytometry. Among the tested cytokines, IL-15 was the most potent cytokine in stimulating tumor cell killing and expanding both natural killer (NK) cells and CD8(+) T cells. Gene-based delivery of IL-15/IL-15Rα to tumor cells, shows expansion of NK cells, activation of NK cells, CD4(+) and CD8(+) T cells, and killing of tumor spheroids. Both NK cells and CD8(+) T cells are necessary for tumor cell killing and CD4(+) T-cell activation was reduced without NK cells. Co-delivery of K2-Fc improved tumor cell killing coinciding with increased activation of NK cells, which was independent of bystander T cells. CD4(+) or CD8(+) T cells were not affected by the co-delivery of K2-Fc even though NK-cell activation impacted CD4(+) T-cell activation. This study demonstrates that gene-based delivery of IL-15/IL-15Rα to tumor cells effectively mediates anti-tumor activity and sensitizes the tumor microenvironment for therapy with αPD-L1 therapeutics mainly by impacting NK cells. These findings warrant further investigation of gene-based IL-15 and K2-Fc delivery in vivo. Nature Publishing Group UK 2023-11-03 /pmc/articles/PMC10624833/ /pubmed/37923822 http://dx.doi.org/10.1038/s41598-023-45948-w Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Awad, Robin Maximilian
De Vlaeminck, Yannick
Meeus, Fien
Ertveldt, Thomas
Zeven, Katty
Ceuppens, Hannelore
Goyvaerts, Cleo
Verdonck, Magali
Salguero, Gustavo
Raes, Geert
Devoogdt, Nick
Breckpot, Karine
In vitro modelling of local gene therapy with IL-15/IL-15Rα and a PD-L1 antagonist in melanoma reveals an interplay between NK cells and CD4(+) T cells
title In vitro modelling of local gene therapy with IL-15/IL-15Rα and a PD-L1 antagonist in melanoma reveals an interplay between NK cells and CD4(+) T cells
title_full In vitro modelling of local gene therapy with IL-15/IL-15Rα and a PD-L1 antagonist in melanoma reveals an interplay between NK cells and CD4(+) T cells
title_fullStr In vitro modelling of local gene therapy with IL-15/IL-15Rα and a PD-L1 antagonist in melanoma reveals an interplay between NK cells and CD4(+) T cells
title_full_unstemmed In vitro modelling of local gene therapy with IL-15/IL-15Rα and a PD-L1 antagonist in melanoma reveals an interplay between NK cells and CD4(+) T cells
title_short In vitro modelling of local gene therapy with IL-15/IL-15Rα and a PD-L1 antagonist in melanoma reveals an interplay between NK cells and CD4(+) T cells
title_sort in vitro modelling of local gene therapy with il-15/il-15rα and a pd-l1 antagonist in melanoma reveals an interplay between nk cells and cd4(+) t cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10624833/
https://www.ncbi.nlm.nih.gov/pubmed/37923822
http://dx.doi.org/10.1038/s41598-023-45948-w
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