Cancer Immunology, Immunotherapy

, Volume 63, Issue 4, pp 321–333

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

  • Christian Bauer
  • Alexander Sterzik
  • Franz Bauernfeind
  • Peter Duewell
  • Claudius Conrad
  • Rosemarie Kiefl
  • Stefan Endres
  • Andreas Eigler
  • Max Schnurr
  • Marc Dauer
Original Article

DOI: 10.1007/s00262-013-1510-y

Cite this article as:
Bauer, C., Sterzik, A., Bauernfeind, F. et al. Cancer Immunol Immunother (2014) 63: 321. doi:10.1007/s00262-013-1510-y

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.

Keywords

Vaccination Dendritic cells Pancreatic carcinoma Chemotherapy Gemcitabine Survival 

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

Supplementary material

262_2013_1510_MOESM1_ESM.pdf (112 kb)
Supplementary material 1 (PDF 112 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Christian Bauer
    • 1
  • Alexander Sterzik
    • 1
  • Franz Bauernfeind
    • 1
  • Peter Duewell
    • 1
  • Claudius Conrad
    • 5
  • Rosemarie Kiefl
    • 1
  • Stefan Endres
    • 2
  • Andreas Eigler
    • 4
  • Max Schnurr
    • 1
  • Marc Dauer
    • 3
  1. 1.Section of Gastroenterology, Medizinische Klinik und Poliklinik IVUniversity of MunichMunichGermany
  2. 2.Division of Clinical Pharmacology, Medizinische Klinik und Poliklinik IVUniversity of MunichMunichGermany
  3. 3.Department of Medicine IIKliniken St. ElisabethNeuburg an der DonauGermany
  4. 4.Department of Internal Medicine IKlinikum Dritter OrdenMunichGermany
  5. 5.Massachusetts General HospitalHarvard UniversityBostonUSA