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2007 Program and Abstracts | 2007 Posters
Dendritic Cells Generation from Peripheral Blood Mononuclear Cells Obtained from Jaundiced Patients with Pancreatic Adenocarcinoma
Andre S. Matheus*1, Mariza Treglia2, Jose Jukemura1, Jose Eduardo M. Cunha1, Marcel C. Machado1, Jose Alexandre M. Barbuto2
1Gastroenterology, University of São Paulo, São Paulo, Brazil; 2Biomedical Science Institute, University of São Paulo, São Paulo, Brazil

Pancreatic adenocarcinoma (PAde) is an aggressive malignancy with poor prognosis, urging for improved or new therapeutic strategies. Dendritic cells (DC) -based vaccination is one of such promising approaches. DC are the most potent antigen-presenting cells and central to the induction and maintenance of an immune response. However, in cancer patients DC generation and function may be deficient, imposing an obstacle to the success of their use. Here, we describe the in vitro generation of DC from peripheral blood mononuclear cells (PBMC) obtained from jaundiced patients with PAde and, also, the effect of jaundiced plasma (JP) in the phenomenon. PBMC were separated from blood obtained from 10 patients and 10 healthy controls over a density gradient. Adherent cells were cultured with GM-CSF and IL-4 (50ng/mL) for 7 days. On the 5th day, TNF-Alpha (50ng/mL) was added for DC activation. Cultures were performed in 10% JP or normal plasma (NP). Non-adherent cells were harvested at day 7, labeled with FITC- or PE-conjugated monoclonal antibodies against CD86, CD80, CD1a, CD11c, CD14, HLA-DR and analyzed by flow cytometry. Patient cells, cultured in 10% JP, compared to healthy donor cells, cultured in 10% NP, had a significantly (p<0.05) lower expression of CD11c, CD80, CD86 and HLA-DR. It is noteworthy that cells generated from patients PBMC did not express CD11c, while 60% of those derived from healthy donor cells did so. The presence of JP in healthy donor cells cultures caused a significant decrease in the percentage of HLA-DR+ (66% x 89%), CD11c+ (43% x 60%) and CD86+ cells (43% x 66%). Finally, when patients PBMC were cultured in NP, a significant increase in HLA-DR (87% x 75%) and a tendency (p=0.06) to increase in CD86 (43% x 30%) expression occurred. These data indicate a significant alteration in the patients PBMC ability to differentiate into DC in vitro, a phenomenon that seems to depend both on soluble factors present in plasma and on the cells, themselves.


2007 Program and Abstracts | 2007 Posters
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