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Prophylactic DNA vaccine targeting Foxp3+ regulatory T cells depletes myeloid-derived suppressor cells and improves anti-melanoma immune responses in a murine model

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Abstract

Regulatory T cells (Treg) and myeloid-derived suppressor cells (MDSC) are the two important and interactive immunosuppressive components of the tumor microenvironment that hamper anti-tumor immune responses. Therefore, targeting these two populations together might be beneficial for overcoming immune suppression in the tumor microenvironment. We have recently shown that prophylactic Foxp3 DNA/recombinant protein vaccine (Foxp3 vaccine) promotes immunity against Treg in tumor-free conditions. In the present study, we investigated the immune modulatory effects of a prophylactic regimen of the redesigned Foxp3 vaccine in the B16F10 melanoma model. Our results indicate that Foxp3 vaccination continuously reduces Treg population in both the tumor site and the spleen. Surprisingly, Treg reduction was associated with a significant decrease in the frequency of MDSC, both in the spleen and in the tumor environment. Furthermore, Foxp3 vaccination resulted in a significant reduction of arginase-1(Arg-1)-induced nitric oxide synthase (iNOS), reactive oxygen species (ROS) and suppressed MDSC activity. Moreover, this concurrent depletion restored production of inflammatory cytokine IFN-γ and enhanced tumor-specific CTL response, which subsequently resulted in the reduction of tumor growth and the improved survival rate of vaccinated mice. In conclusion, our results revealed that Foxp3 vaccine promotes an immune response against tumor by targeting both Treg and MDSC, which could be exploited as a potential immunotherapy approach.

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Abbreviations

Arg-1:

Arginase-1

BCA:

Bicinchoninic acid

CFSE:

Carboxy fluorescein succinimidyl ester

DCFDA:

2′,7′-Dichlorodihydrofluorescein diacetate

E. coli :

Escherichia coli

Gr-1:

Granulocyte differentiation marker 1

His-tag:

(Poly)-histidine-tagged

IFA:

Incomplete Freund’s adjuvant

iNOS:

Induced nitric oxide synthase

MDSC:

Myeloid-derived suppressor cell(s)

Ni–NTA:

Nickel nitrilotriacetic acid

ROS:

Reactive oxygen species

Treg:

Regulatory T cell(s)

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Author information

Authors and Affiliations

  1. Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

    Afshin Namdar & Abbas Rezaei

  2. Department of Immunology, Building No. 7, School of Medicine, Tehran University of Medical Sciences, Poursina Avenue, Tehran, 14155-6447, Iran

    Afshin Namdar, Reza Mirzaei, Roobina Boghosian, Hamid Reza Mirzaei, Hamid Farajifard, Mehdi Zavar, Farshid Noorbakhsh & Jamshid Hadjati

  3. Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran

    Arash Memarnejadian

  4. Recurrent Abortion Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

    Morteza Samadi

  5. Department of Clinical Laboratory Sciences, School of Allied Medical Sciences, Kashan University of Medical Sciences, Kashan, Iran

    Hamid Reza Mirzaei

  6. Department of Virology, Pasteur Institute of Iran, Tehran, Iran

    Kayhan Azadmanesh

  7. Department of Dentistry, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Canada

    Afshin Namdar & Shokrollah Elahi

  8. Department of Medical Microbiology and Immunology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Canada

    Shokrollah Elahi

  9. Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    Hamid Reza Mirzaei

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  1. Afshin Namdar
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  2. Reza Mirzaei
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Contributions

AN performed plasmid and protein preparation, animal studies, flow cytometry analysis, real-time PCR, suppression assay, and nitrite assay, analyzed the data, and wrote the manuscript. RM contributed in flow cytometry analysis and animal studies. AM advised in plasmid design. RB performed intracellular cytokine staining. MS assisted in plasmid and protein preparation. HRM performed Treg staining and CD107a cytotoxic assay. HF performed RNA extraction. MZ performed MDSC staining. KA provided the main construct and co-supervised the designing of plasmids. SE contributed to writing and editing the article. FN advised on plasmid design and data analysis. AR co-supervised the study. JH supervised the study.

Corresponding author

Correspondence to Jamshid Hadjati.

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Funding

The main part of this study was supported by the Iran National Science Foundation (INSF, Grant Number 90007957) and partially by the Isfahan University of Medical Sciences (Grant Number 391146).

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

Mouse experiments were approved by the animal ethics committee of Tehran University of Medical Sciences (Tehran, Iran).

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Namdar, A., Mirzaei, R., Memarnejadian, A. et al. Prophylactic DNA vaccine targeting Foxp3+ regulatory T cells depletes myeloid-derived suppressor cells and improves anti-melanoma immune responses in a murine model. Cancer Immunol Immunother 67, 367–379 (2018). https://doi.org/10.1007/s00262-017-2088-6

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