Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer-0

After transduction, DC were cultured up to 48 h. GFP expression was analyzed by flow cytometry at 24 h () and 48 h culture (), as described in . A population of GFPCD86DC at 24 h () and 48 h () is shown. Staining with isotype control is indicated in and . The percentage of GFPCD86DC is indicated in the upper right corner, and MFI is also reported for D and C. The density plots for one representative experiment of four are shown.<p><b>Copyright information:</b></p><p>Taken from "Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer"</p><p>http://www.translational-medicine.com/content/6/1/38</p><p>Journal of Translational Medicine 2008;6():38-38.</p><p>Published online 22 Jul 2008</p><p>PMCID:PMC2507704.</p><p></p

Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer-2

DC. A population of DXCD86obtained at 4°C and at 37°C is shown. Values indicate the percentage and MFI of DXCD86. Density plots of a representative experiment of three are shown.<p><b>Copyright information:</b></p><p>Taken from "Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer"</p><p>http://www.translational-medicine.com/content/6/1/38</p><p>Journal of Translational Medicine 2008;6():38-38.</p><p>Published online 22 Jul 2008</p><p>PMCID:PMC2507704.</p><p></p

Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer-4

DC, or CV-DC were seeded into a 48 well plate in 1 ml of culture medium in the presence or absence of the indicated stimuli. For IL-12 assays, cells were stimulated with IFN-γ + LPS, and for IP-10 and MIG assays cells were stimulated with LPS only. The supernatants from stimulated and unstimulated DC were harvested after 48 h culture. IL-12p), IP-10 , and MIG were measured by ELISA. Values refer to cytokine concentration expressed in ng/million cells. One representative experiment of four is shown.<p><b>Copyright information:</b></p><p>Taken from "Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer"</p><p>http://www.translational-medicine.com/content/6/1/38</p><p>Journal of Translational Medicine 2008;6():38-38.</p><p>Published online 22 Jul 2008</p><p>PMCID:PMC2507704.</p><p></p

Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer-3

Ell proliferation. The effect of pulsing 100% (DC KLH 100 and AdDC KLH 100) or 30% of cells (AdDC KLH 30) with 10 μg/ml KLH prior to initiating the MLR assay was assessed. After pre-treatment with mitomycin C and KLH, where indicated, DC and CCL21-DC were mixed with allogeneic T cells at several ratios, as shown. Cell proliferation was assessed by BrdU incorporation following 5 days incubation at 37°C. One representative experiment of three is shown. T cells were cultured for 6 days with IL-2 (15 U/ml) prior to the TT presentation assay. After mitomycin-C treatment and KLH pulsing where indicated, DC (), CCL21-DC (), or CV-DC () were mixed at 1:10 ratio to autologous T cells in the presence of TT (2 μg/ml) or BSA (2 μg/ml). In (), IFN-γ production was measured in the cell supernatant collected after 48 hours of cell proliferation. The measured values of IFN-γ detected by ELISA were within the linear portion of the IFN-γ concentration curve. The concentration of IFN-γ is expressed in pg/ml with basal IFN-γ production subtracted. In (), T cell proliferation in response to TT and BSA was measured by BrdU incorporation following 5 days co-culture at 37°C and is expressed as a percentage of proliferating cells. One representative experiment of at least three is shown. P value ≤ 0.05 was considered significant.<p><b>Copyright information:</b></p><p>Taken from "Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer"</p><p>http://www.translational-medicine.com/content/6/1/38</p><p>Journal of Translational Medicine 2008;6():38-38.</p><p>Published online 22 Jul 2008</p><p>PMCID:PMC2507704.</p><p></p