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. Author manuscript; available in PMC: 2012 Feb 1.
Published in final edited form as: Prostate. 2010 Aug 17;71(2):125–133. doi: 10.1002/pros.21228

Fig. 1. Feasibility of generating tumor-loaded functional αDC1s from prostate cancer patients.

Fig. 1

A. Recovery of DCs is not affected by tumor loading. Monocytes isolated from PCa patients' blood were plated at 5×105 cells per well in 24 well plates and cultured for 6 days in the presence of GM-CSF and IL4. On day 6, DC were induced to mature into polarized αDC1 (with TNFα, IL1β, IFNα, IFNγ, and Poly-I:C) or sDC (with TNFα, IL1β, IL6, and PGE2) in the presence or absence of UV-irradiated apoptotic LNCaP cells. αDC1s and sDCs were harvested on day 8, washed and counted under trypan blue. DC recovery was calculated as % of plated monocytes (n=10). B-C. Intact mature phenotype of the αDC1s and sDCs loaded with UVB-irradiated LNCaP cells. αDC1s and sDCs were harvested on day 8 and analyzed by flow cytometry for the expression of maturation and migratory markers. B. Representative data from one patient. C. Cumulative data from 10 donors expressed as mean of delta MFIs (increase in specific fluorescence over isotype controls) ±SD; n=10. Note that the expression levels of CD86 and CD80 on αDC1s were higher than those on sDCs (P < 0.05), while the expression levels of CD11c, CD83 and CCR7 were comparable. In addition, there was no significant impact of the tumor loading in the expression of these molecules in DCs by the pulsing of tumor antigens. C. Elevated L-12 production by αDC1s is not affected by loading with PCa cells. αDC1s and sDCs from 10 PCa patients were harvested on day 8 and stimulated with CD40L for 24 hr. The αDC1s showed significantly higher production of IL-12p70 than sDCs (P < 0.001). Loading of tumor antigen onto αDC1s did not significantly changed IL-12 secretion by these cells (n=10 different patients).