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01.06.2008 | Original Article

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

verfasst von: Peter Dubsky, Hubert Hayden, Monika Sachet, Thomas Bachleitner-Hofmann, Michaela Hassler, Roswitha Pfragner, Michael Gnant, Anton Stift, Josef Friedl

Erschienen in: Cancer Immunology, Immunotherapy | Ausgabe 6/2008

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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 CD1a^high, CD14^low 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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Titel: Allogeneic tumor lysate can serve as both antigen source and protein supplementation for dendritic cell culture
verfasst von: Peter Dubsky
Hubert Hayden
Monika Sachet
Thomas Bachleitner-Hofmann
Michaela Hassler
Roswitha Pfragner
Michael Gnant
Anton Stift
Josef Friedl
Publikationsdatum: 01.06.2008
Verlag: Springer-Verlag
Erschienen in: Cancer Immunology, Immunotherapy / Ausgabe 6/2008
Print ISSN: 0340-7004
Elektronische ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-007-0422-0

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