Tetracyclines enhance antitumor T-cell immunity via the Zap70 signaling pathway

Background Cancer immunotherapy including immune checkpoint inhibitors is only effective for a limited population of patients with cancer. Therefore, the development of novel cancer immunotherapy is anticipated. In preliminary studies, we demonstrated that tetracyclines enhanced T-cell responses. Th...

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Main Authors: Atsushi Kumanogoh, Hisashi Wada, Yoshito Takeda, Mari Tone, Yasushi Shintani, Kota Iwahori, Michinari Hirata, Azumi Ueyama, Akiyoshi Tani, Jun-Ichi Haruta
Format: Article
Language:English
Published: BMJ Publishing Group 2024-04-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/12/4/e008334.full
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author Atsushi Kumanogoh
Hisashi Wada
Yoshito Takeda
Mari Tone
Yasushi Shintani
Kota Iwahori
Michinari Hirata
Azumi Ueyama
Akiyoshi Tani
Jun-Ichi Haruta
author_facet Atsushi Kumanogoh
Hisashi Wada
Yoshito Takeda
Mari Tone
Yasushi Shintani
Kota Iwahori
Michinari Hirata
Azumi Ueyama
Akiyoshi Tani
Jun-Ichi Haruta
author_sort Atsushi Kumanogoh
collection DOAJ
description Background Cancer immunotherapy including immune checkpoint inhibitors is only effective for a limited population of patients with cancer. Therefore, the development of novel cancer immunotherapy is anticipated. In preliminary studies, we demonstrated that tetracyclines enhanced T-cell responses. Therefore, we herein investigated the efficacy of tetracyclines on antitumor T-cell responses by human peripheral T cells, murine models, and the lung tumor tissues of patients with non-small cell lung cancer (NSCLC), with a focus on signaling pathways in T cells.Methods The cytotoxicity of peripheral and lung tumor-infiltrated human T cells against tumor cells was assessed by using bispecific T-cell engager (BiTE) technology (BiTE-assay system). The effects of tetracyclines on T cells in the peripheral blood of healthy donors and the tumor tissues of patients with NSCLC were examined using the BiTE-assay system in comparison with anti-programmed cell death-1 (PD-1) antibody, nivolumab. T-cell signaling molecules were analyzed by flow cytometry, ELISA, and qRT-PCR. To investigate the in vivo antitumor effects of tetracyclines, tetracyclines were administered orally to BALB/c mice engrafted with murine tumor cell lines, either in the presence or absence of anti-mouse CD8 inhibitors.Results The results obtained revealed that tetracyclines enhanced antitumor T-cell cytotoxicity with the upregulation of granzyme B and increased secretion of interferon-γ in human peripheral T cells and the lung tumor tissues of patients with NSCLC. The analysis of T-cell signaling showed that CD69 in both CD4+ and CD8+ T cells was upregulated by minocycline. Downstream of T-cell receptor signaling, Zap70 phosphorylation and Nur77 were also upregulated by minocycline in the early phase after T-cell activation. These changes were not observed in T cells treated with anti-PD-1 antibodies under the same conditions. The administration of tetracyclines exhibited antitumor efficacy with the upregulation of CD69 and increases in tumor antigen-specific T cells in murine tumor models. These changes were canceled by the administration of anti-mouse CD8 inhibitors.Conclusions In conclusion, tetracyclines enhanced antitumor T-cell immunity via Zap70 signaling. These results will contribute to the development of novel cancer immunotherapy.
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series Journal for ImmunoTherapy of Cancer
spelling doaj-art-c59908baeae14bb5b96141a34ab1d9492024-11-14T02:40:10ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262024-04-0112410.1136/jitc-2023-008334Tetracyclines enhance antitumor T-cell immunity via the Zap70 signaling pathwayAtsushi Kumanogoh0Hisashi Wada1Yoshito Takeda2Mari Tone3Yasushi Shintani4Kota Iwahori5Michinari Hirata6Azumi Ueyama7Akiyoshi Tani8Jun-Ichi Haruta91 Department of Respiratory Medicine and Clinical Immunology, Osaka University Graduate School of Medicine, Suita City, JapanDepartment of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Osaka, JapanDepartment of Respiratory Medicine and Clinical Immunology, Graduate School of Medicine, Osaka University, Osaka, JapanDepartment of Respiratory Medicine and Clinical Immunology, Graduate School of Medicine, Osaka University, Osaka, JapanDepartment of General Thoracic Surgery, Graduate School of Medicine, Osaka University, Osaka, JapanDepartment of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Osaka, JapanDepartment of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Osaka, JapanDepartment of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Osaka, JapanCompound Library Screening Center, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, JapanLead Explorating Units, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, JapanBackground Cancer immunotherapy including immune checkpoint inhibitors is only effective for a limited population of patients with cancer. Therefore, the development of novel cancer immunotherapy is anticipated. In preliminary studies, we demonstrated that tetracyclines enhanced T-cell responses. Therefore, we herein investigated the efficacy of tetracyclines on antitumor T-cell responses by human peripheral T cells, murine models, and the lung tumor tissues of patients with non-small cell lung cancer (NSCLC), with a focus on signaling pathways in T cells.Methods The cytotoxicity of peripheral and lung tumor-infiltrated human T cells against tumor cells was assessed by using bispecific T-cell engager (BiTE) technology (BiTE-assay system). The effects of tetracyclines on T cells in the peripheral blood of healthy donors and the tumor tissues of patients with NSCLC were examined using the BiTE-assay system in comparison with anti-programmed cell death-1 (PD-1) antibody, nivolumab. T-cell signaling molecules were analyzed by flow cytometry, ELISA, and qRT-PCR. To investigate the in vivo antitumor effects of tetracyclines, tetracyclines were administered orally to BALB/c mice engrafted with murine tumor cell lines, either in the presence or absence of anti-mouse CD8 inhibitors.Results The results obtained revealed that tetracyclines enhanced antitumor T-cell cytotoxicity with the upregulation of granzyme B and increased secretion of interferon-γ in human peripheral T cells and the lung tumor tissues of patients with NSCLC. The analysis of T-cell signaling showed that CD69 in both CD4+ and CD8+ T cells was upregulated by minocycline. Downstream of T-cell receptor signaling, Zap70 phosphorylation and Nur77 were also upregulated by minocycline in the early phase after T-cell activation. These changes were not observed in T cells treated with anti-PD-1 antibodies under the same conditions. The administration of tetracyclines exhibited antitumor efficacy with the upregulation of CD69 and increases in tumor antigen-specific T cells in murine tumor models. These changes were canceled by the administration of anti-mouse CD8 inhibitors.Conclusions In conclusion, tetracyclines enhanced antitumor T-cell immunity via Zap70 signaling. These results will contribute to the development of novel cancer immunotherapy.https://jitc.bmj.com/content/12/4/e008334.full
spellingShingle Atsushi Kumanogoh
Hisashi Wada
Yoshito Takeda
Mari Tone
Yasushi Shintani
Kota Iwahori
Michinari Hirata
Azumi Ueyama
Akiyoshi Tani
Jun-Ichi Haruta
Tetracyclines enhance antitumor T-cell immunity via the Zap70 signaling pathway
Journal for ImmunoTherapy of Cancer
title Tetracyclines enhance antitumor T-cell immunity via the Zap70 signaling pathway
title_full Tetracyclines enhance antitumor T-cell immunity via the Zap70 signaling pathway
title_fullStr Tetracyclines enhance antitumor T-cell immunity via the Zap70 signaling pathway
title_full_unstemmed Tetracyclines enhance antitumor T-cell immunity via the Zap70 signaling pathway
title_short Tetracyclines enhance antitumor T-cell immunity via the Zap70 signaling pathway
title_sort tetracyclines enhance antitumor t cell immunity via the zap70 signaling pathway
url https://jitc.bmj.com/content/12/4/e008334.full
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