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Fms-like tyrosine kinase 3 receptor ligand (Flt3L)-based vaccination administered with an adenoviral vector prevents tumor growth of colorectal cancer in a BALB/c mouse model

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Abstract

Purpose

Colorectal cancer is the third most frequent cancer in industrial nations. Therapeutic strategies to treat metastatic disease and prevent recurrence are needed. Anti-tumor immunity can be induced by dendritic cells. Dendritic cells can be expanded by the fms-like tyrosine kinase 3 ligand (Flt3L) in vivo. The aim of this study was to develop an adenoviral-based immune-gene therapy of colorectal cancer with Flt3L in a BALB/c mouse model.

Methods

A new Flt3L-encoding adenoviral vector (pAdFlt3L) was administered in two approaches in a CT26 colon cancer model in female BALB/c mice. In the therapeutic approach, pAdFlt3L was injected into the tail vein or directly into subcutaneous CT26 colon carcinoma tumors in BALB/c mice. In the vaccination protocol, mice were vaccinated with CT26 cell lysate and pAdFlt3L subcutaneous prior to subcutaneous application of vital CT26 cells.

Results

Application of pAdFlt3L led to high levels of Flt3L in vitro and in vivo. Significant expansion of dendritic cells after application of pAdFlt3L in vivo was confirmed by the use of CD11c and CD11b surface markers in immunohistochemistry and flow cytometry (p = 0.019). In the therapeutic approach, neither intravenous nor intratumoral treatments with pAdFlt3L lead to regression of CT26 tumors. In the vaccination protocol, vaccination completely prevented tumor growth and resulted in superior survival compared to control mice (p < 0.001).

Conclusions

Our results demonstrate that immunostimulatory therapy with pAdFlt3L is effective to prevent tumor development through vaccination and may represent a therapeutic tool to prevent metastatic disease.

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Abbreviations

BSA:

Bovine serum albumin

CT26 cells:

Colon carcinoma cell line

DC:

Dendritic cells

DMEM:

Dulbecco’s modified Eagle medium supplemented with 10 % heat-inactivated fetal bovine serum

Flt3L:

Fms-like tyrosine kinase 3 receptor ligand

FCS:

Heat-inactivated fetal bovine serum

FITC:

Fluorescein isothiocyanate

FACS:

Fluorscence activated cell sorter flow cytometry

GM-CSF:

Granulocyte macrophage colony stimulating factor

GFP:

Green fluorescent protein (GFP)

HEK-293 cells:

Human embryonic kidney cell line

MOI:

Multiplicity of infection

pAdFlt3L:

Replication-deficient adenoviral vector encoding for Flt3L and GFP

pAdGFP:

Replication-deficient adenoviral vector encoding for GFP

PBS:

Phosphate-buffered saline

RIPA-buffer:

50, 150 mM sodiumchloride, 1 % Nonidet p-40, 0.5 % sodiumdeoxycholate, 1 % sodium dodecyl sulfate

Rpm:

Runs per minute

SDS:

Sodium dodecyl sulfate-plyacrylamide

TBS:

Tris-buffered saline

TBS Tween:

TRis-biffered saline Tween

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Acknowledgments

The authors thank Stefan Urban and Anke Lonsdorf for helpful discussion. We also thank Ulrike Protzer for kindly providing pAdGFP and Sabine Tuma and Jasmin Geib for their excellent technical assistance.

Conflict of interest

None.

Author information

Author notes

  1. Carina Riediger

    Present address: Department of General, Thoracic and Vascular Surgery, University Hospital Carl Gustav Carus, Technische Universität Dresden, Fetscherstrasse 74, 01307, Dresden, Germany

  2. Gerhard Wingender

    Present address: Division of Developmental Immunology, La Jolla Institute for Allergy and Immunology, 9420 Athena Circle, La Jolla, CA, 92037, USA

Authors and Affiliations

  1. Department of Internal Medicine IV and Otto-Meyerhof-Center for Medical Sciences, University of Heidelberg, Heidelberg, Germany

    Carina Riediger, Wolfgang Stremmel & Jens Encke

  2. Institute of Molecular Medicine and Experimental Immunology, University of Bonn, Bonn, Germany

    Gerhard Wingender & Percy Knolle

  3. Institute of Pathology, University of Heidelberg, Heidelberg, Germany

    Sebastian Aulmann

  4. Institute of Molecular Immunology, Technische Universität München, Munich, Germany

    Percy Knolle

Authors
  1. Carina Riediger
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  2. Gerhard Wingender
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Corresponding author

Correspondence to Carina Riediger.

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Riediger, C., Wingender, G., Knolle, P. et al. Fms-like tyrosine kinase 3 receptor ligand (Flt3L)-based vaccination administered with an adenoviral vector prevents tumor growth of colorectal cancer in a BALB/c mouse model. J Cancer Res Clin Oncol 139, 2097–2110 (2013). https://doi.org/10.1007/s00432-013-1532-z

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  • DOI: https://doi.org/10.1007/s00432-013-1532-z

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