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Concomitant gemcitabine therapy negatively affects DC vaccine-induced CD8+ T-cell and B-cell responses but improves clinical efficacy in a murine pancreatic carcinoma model

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Abstract

Background

Multiple studies have shown that dendritic cell (DC)-based vaccines can induce antitumor immunity. Previously, we reported that gemcitabine enhances the efficacy of DC vaccination in a mouse model of pancreatic carcinoma. The present study aimed at investigating the influence of gemcitabine on vaccine-induced anti-tumoral immune responses in a syngeneic pancreatic cancer model.

Materials and methods

Subcutaneous or orthotopic pancreatic tumors were induced in C57BL/6 mice using Panc02 cells expressing the model antigen OVA. Bone marrow-derived DC were loaded with soluble OVA protein (OVA-DC). Animals received gemcitabine twice weekly. OVA-specific CD8+ T-cells and antibody titers were monitored by FACS analysis and ELISA, respectively.

Results

Gemcitabine enhanced clinical efficacy of the OVA-DC vaccine. Interestingly, gemcitabine significantly suppressed the vaccine-induced frequency of antigen-specific CD8+ T-cells and antibody titers. DC migration to draining lymph nodes and antigen cross-presentation were unaffected. Despite reduced numbers of tumor-reactive T-cells in peripheral blood, in vivo cytotoxicity assays revealed that cytotoxic T-cell (CTL)-mediated killing was preserved. In vitro assays revealed sensitization of tumor cells to CTL-mediated lysis by gemcitabine. In addition, gemcitabine facilitated recruitment of CD8+ T-cells into tumors in DC-vaccinated mice. T- and B-cell suppression by gemcitabine could be avoided by starting chemotherapy after two cycles of DC vaccination.

Conclusions

Gemcitabine enhances therapeutic efficacy of DC vaccination despite its negative influence on vaccine-induced T-cell proliferation. Quantitative analysis of tumor-reactive T-cells in peripheral blood may thus not predict vaccination success in the setting of concomitant chemotherapy.

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Abbreviations

CFSE:

Carboxyfluorescein succinimidyl ester

CTL:

Cytotoxic T-cell

DC:

Dendritic cell

ELISA:

Enzyme-linked immunosorbent assay

FACS:

Fluorescence-activated cell sorter

FoxP3:

Forkheadbox P3

Gem:

Gemcitabine

GM-CSF:

Granulocyte macrophage colony-stimulating factor

IFN-γ:

Interferon-γ

i.v.:

Intravenous

i.p.:

Intraperitoneal

ICS:

Intracellular staining

LPS:

Lipopolysaccharide

MHC-I:

Major histocompatibility complex I

MDSC:

Myeloid-derived suppressor cell

OD:

Optical density

OVA:

Ovalbumine

OVA-DC:

OVA protein-loaded DC

p15E:

Retroviral protein expressed by Panc02 cells

s.c.:

Subcutaneous

SIINFEKL:

Immunodominant MHC-I epitope of the ovalbumine protein

TNF-α:

Tumor necrosis factor-α

Treg :

Regulatory CD4+ Foxp3+ T-cell

TRP2:

Tryosinase-related peptide 2

U-DC:

Unloaded but LPS-stimulated DC

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Acknowledgments

Christian Bauer, Franz Bauernfeind and Marc Dauer were supported by grants from the University of Munich (FoeFoLe No. 481, Promotionsstudium Molekulare Medizin, Gravenhorst-Stiftung) and the Saarland University (HOMFOR). Max Schnurr was supported by the Deutsche Krebshilfe (Max Eder Research Grant) and the Deutsche Forschungsgemeinschaft (SCHN 664/3-1, SCHN 664/3-2 and GK 1202). Transgenic OT-1 animals were kindly provided by Prof. Thomas Brocker (Department of Immunology, University of Munich, Germany).

Conflict of interest

All authors declare that no conflicts of interest exist.

Author information

Authors and Affiliations

  1. Section of Gastroenterology, Medizinische Klinik und Poliklinik IV, University of Munich, Munich, Germany

    Christian Bauer, Alexander Sterzik, Franz Bauernfeind, Peter Duewell, Rosemarie Kiefl & Max Schnurr

  2. Division of Clinical Pharmacology, Medizinische Klinik und Poliklinik IV, University of Munich, Munich, Germany

    Stefan Endres

  3. Department of Medicine II, Kliniken St. Elisabeth, Müller-Gnadenegg-Weg 4, 86633, Neuburg an der Donau, Germany

    Marc Dauer

  4. Department of Internal Medicine I, Klinikum Dritter Orden, Munich, Germany

    Andreas Eigler

  5. Massachusetts General Hospital, Harvard University, Boston, MA, USA

    Claudius Conrad

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  1. Christian Bauer
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  2. Alexander Sterzik
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  3. Franz Bauernfeind
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Correspondence to Marc Dauer.

Additional information

Christian Bauer and Alexander Sterzik, and Max Schnurr and Marc Dauer have contributed equally to this article.

This work is part of the doctoral thesis of Alexander Sterzik and Franz Bauernfeind at the University of Munich, Germany.

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Bauer, C., Sterzik, A., Bauernfeind, F. et al. Concomitant gemcitabine therapy negatively affects DC vaccine-induced CD8+ T-cell and B-cell responses but improves clinical efficacy in a murine pancreatic carcinoma model. Cancer Immunol Immunother 63, 321–333 (2014). https://doi.org/10.1007/s00262-013-1510-y

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