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


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 IFNgamma in CD8+T-cells of vaccinated MTC patients. Both types of DC induced killing of SHER-I after short in vitro restimulation. 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.
Dubsky
Peter
P
Department of Surgery, Medical University of Vienna, Währinger Gürtel 18-20, 1090 Vienna, Austria. peter.dubsky@meduniwien.ac.at
Hayden
Hubert
H
Sachet
Monika
M
Bachleitner-Hofmann
Thomas
T
Hassler
Michaela
M
Pfragner
Roswitha
R
Gnant
Michael
M
Stift
Anton
A
Friedl
Josef
J
D000951:Antigens, Neoplasm / Q000737:chemistry*; D017252:CD3 Complex / Q000096:biosynthesis; D002277:Carcinoma / Q000276:immunology* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D045744:Cell Line, Tumor; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D003713:Dendritic Cells / Q000166:cytology*; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D016753:Interleukin-10 / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D009369:Neoplasms / Q000276:immunology*; D011506:Proteins / Q000737:chemistry*; D013964:Thyroid Neoplasms / Q000276:immunology* / Q000378:metabolism; D014184:Transplantation, Homologous / Q000379:methods*
D000951:Antigens, Neoplasm; D017252:CD3 Complex; D016895:Culture Media, Serum-Free; D011506:Proteins; D016753:Interleukin-10; D015847:Interleukin-4