Intratumoral expression of IL-12 and CD40 ligand (CD154) from plasmids generates antitumor responses that eliminate tumoral T regs

Abstract Intratumoral immunotherapy (ITIT) strives to generate effective antitumor immunity by directly stimulating the immune system in tumors to reverse local tumor-mediated immune suppression. In vivo expression of Interleukin-12 (IL-12) using in vivo plasmid transfection as an intratumoral cance...

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Main Authors: Gregory W. Ho, Alicia Santos, Jennifer Fields, Pamela C. Rosato, Mary Jo Turk, Nicole F. Steinmetz, Steven Fiering
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-025-09535-5
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author Gregory W. Ho
Alicia Santos
Jennifer Fields
Pamela C. Rosato
Mary Jo Turk
Nicole F. Steinmetz
Steven Fiering
author_facet Gregory W. Ho
Alicia Santos
Jennifer Fields
Pamela C. Rosato
Mary Jo Turk
Nicole F. Steinmetz
Steven Fiering
author_sort Gregory W. Ho
collection DOAJ
description Abstract Intratumoral immunotherapy (ITIT) strives to generate effective antitumor immunity by directly stimulating the immune system in tumors to reverse local tumor-mediated immune suppression. In vivo expression of Interleukin-12 (IL-12) using in vivo plasmid transfection as an intratumoral cancer immunotherapy entered Phase II clinical trials for metastatic melanoma but to limited clinical success. We sought to improve the efficacy of in vivo IL-12 electroporation by the addition of a CD154 (CD40 ligand)- expressing plasmid to the IL-12 encoding plasmid treatment and assessing efficacy against solid tumors. Mice with intradermal B16F10 melanoma or MC38 murine colon carcinoma tumors received 2 weekly intratumoral (IT) injections of plasmids encoding IL-12 and CD154, followed by in vivo electroporation. The addition of CD154 to IL-12 was superior to IL-12 alone and resulted in frequent tumor clearance of treated tumors, marked by an increase in CD8 T cells and a drastic reduction in T regulatory cells in the tumor microenvironment. Tumor treatment responses were abrogated in mice which lack conventional DC1 cells (BatF3 KO) or lack CD8 T cells. These findings highlight the potential of adding CD154 to IL-12 plasmid electroporation as a cancer immunotherapy and suggest that other combinations would be therapeutically valuable.
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spelling doaj-art-bccb0ac6388e44d7b3b5e944de222b452025-08-20T03:05:25ZengNature PortfolioScientific Reports2045-23222025-07-0115111310.1038/s41598-025-09535-5Intratumoral expression of IL-12 and CD40 ligand (CD154) from plasmids generates antitumor responses that eliminate tumoral T regsGregory W. Ho0Alicia Santos1Jennifer Fields2Pamela C. Rosato3Mary Jo Turk4Nicole F. Steinmetz5Steven Fiering6Department of Microbiology and Immunology, Dartmouth Geisel School of MedicineDepartment of Microbiology and Immunology, Dartmouth Geisel School of MedicineDepartment of Microbiology and Immunology, Dartmouth Geisel School of MedicineDepartment of Microbiology and Immunology, Dartmouth Geisel School of MedicineDepartment of Microbiology and Immunology, Dartmouth Geisel School of MedicineAiiso Yufeng Li Family Department of Chemical and Nano Engineering, University of California San DiegoDepartment of Microbiology and Immunology, Dartmouth Geisel School of MedicineAbstract Intratumoral immunotherapy (ITIT) strives to generate effective antitumor immunity by directly stimulating the immune system in tumors to reverse local tumor-mediated immune suppression. In vivo expression of Interleukin-12 (IL-12) using in vivo plasmid transfection as an intratumoral cancer immunotherapy entered Phase II clinical trials for metastatic melanoma but to limited clinical success. We sought to improve the efficacy of in vivo IL-12 electroporation by the addition of a CD154 (CD40 ligand)- expressing plasmid to the IL-12 encoding plasmid treatment and assessing efficacy against solid tumors. Mice with intradermal B16F10 melanoma or MC38 murine colon carcinoma tumors received 2 weekly intratumoral (IT) injections of plasmids encoding IL-12 and CD154, followed by in vivo electroporation. The addition of CD154 to IL-12 was superior to IL-12 alone and resulted in frequent tumor clearance of treated tumors, marked by an increase in CD8 T cells and a drastic reduction in T regulatory cells in the tumor microenvironment. Tumor treatment responses were abrogated in mice which lack conventional DC1 cells (BatF3 KO) or lack CD8 T cells. These findings highlight the potential of adding CD154 to IL-12 plasmid electroporation as a cancer immunotherapy and suggest that other combinations would be therapeutically valuable.https://doi.org/10.1038/s41598-025-09535-5
spellingShingle Gregory W. Ho
Alicia Santos
Jennifer Fields
Pamela C. Rosato
Mary Jo Turk
Nicole F. Steinmetz
Steven Fiering
Intratumoral expression of IL-12 and CD40 ligand (CD154) from plasmids generates antitumor responses that eliminate tumoral T regs
Scientific Reports
title Intratumoral expression of IL-12 and CD40 ligand (CD154) from plasmids generates antitumor responses that eliminate tumoral T regs
title_full Intratumoral expression of IL-12 and CD40 ligand (CD154) from plasmids generates antitumor responses that eliminate tumoral T regs
title_fullStr Intratumoral expression of IL-12 and CD40 ligand (CD154) from plasmids generates antitumor responses that eliminate tumoral T regs
title_full_unstemmed Intratumoral expression of IL-12 and CD40 ligand (CD154) from plasmids generates antitumor responses that eliminate tumoral T regs
title_short Intratumoral expression of IL-12 and CD40 ligand (CD154) from plasmids generates antitumor responses that eliminate tumoral T regs
title_sort intratumoral expression of il 12 and cd40 ligand cd154 from plasmids generates antitumor responses that eliminate tumoral t regs
url https://doi.org/10.1038/s41598-025-09535-5
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