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Allogeneic tumor lysate can serve as both antigen source and protein supplementation for dendritic cell culture

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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Background

Recent preclinical and clinical evidence suggests the use of allogeneic tumor as a source of antigen for DC-based immunotherapy against cancer. We hypothesized that addition of allogeneic tumor lysate to monocyte-derived DC culture could serve a dual purpose: (1) antigen source and (2) protein supplementation of DC culture media. Protein supplementation whether of known origin (human serum/plasma, fetal bovine serum, human serum albumin) or undeclared origin (“serum-free” media) is a source of variability and bias. We addressed the question whether protein supplementation can be omitted in the presence of allogeneic tumor lysate.

Materials and methods

Human DC cultured in the presence of lysate from medullary thyroid carcinoma (MTC) cell line SHER-I (TuLy-DC) and DC pulsed with the same lysate but cultured in the presence of FBS (FBS-DC) were assessed for morphology, phenotype, maturation and functional properties.

Results

In comparison of FBS-DC/TuLy-DC no significant differences in morphology, phenotype and maturation could be detected. Both culture conditions produced CD1ahigh, CD14low DC with high expression of costimulatory molecules and CD83 upon stimulation. TuLy-DC gave significantly better yields and produced more IL12p70. DC showed high (allo)stimulatory capacity toward T-cells. TuLy-DC induced more intracellular IFNγ in CD8+T-cells of vaccinated MTC patients. Both types of DC induced killing of SHER-I after short in vitro restimulation.

Summary and conclusion

Tumor lysate from SHER-I can substitute for further protein supplementation in DC culture. Allogeneic tumor lysates should be taken into consideration as both source of antigen and protein supplementation in monocyte-derived DC culture.

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Acknowledgments

Special thanks to Prof. Bruno Niederle and Dr. Christian Scheuba for continued support and referral of patients. Prof. Christine Brostjan for discussions and proof reading of the manuscript. We would like to thank our patients for participating in clinical trials and putting continued trust in our work. This work was supported in part by the Funds for Innovative Interdisciplinary Cancer Research of the City of Vienna and by a Research Grant from the DDr. Karl Fellinger Foundation for Cancer Research. This work was performed under the auspices of the Center of Excellence for Clinical and Experimental Oncology University of Vienna.

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Authors and Affiliations

  1. Department of Surgery, Medical University of Vienna, Währinger Gürtel 18-20, 1090, Vienna, Austria

    Peter Dubsky, Hubert Hayden, Monika Sachet, Thomas Bachleitner-Hofmann, Michaela Hassler, Michael Gnant, Anton Stift & Josef Friedl

  2. Department of Pathophysiology, Medical University of Graz, 8010, Graz, Austria

    Roswitha Pfragner

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  1. Peter Dubsky
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  2. Hubert Hayden
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Correspondence to Peter Dubsky.

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Peter Dubsky and Hubert Hayden contributed equally to the manuscript.

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Dubsky, P., Hayden, H., Sachet, M. et al. Allogeneic tumor lysate can serve as both antigen source and protein supplementation for dendritic cell culture. Cancer Immunol Immunother 57, 859–870 (2008). https://doi.org/10.1007/s00262-007-0422-0

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  • DOI: https://doi.org/10.1007/s00262-007-0422-0

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