Feasibility to Generate Monocyte-Derived Dendritic Cell from Coculture with Melanoma Tumor Cells in the Presence of Granulocyte / Macrophage Colony-Stimulating Factor (GM-CSF) and Interleukin-4
Article first published online: 25 MAR 2003
American Journal of Reproductive Immunology
Volume 49, Issue 4, pages 230–238, April 2003
How to Cite
Kim, Y. T., Hersh, E. M. and Trevor, K. T. (2003), Feasibility to Generate Monocyte-Derived Dendritic Cell from Coculture with Melanoma Tumor Cells in the Presence of Granulocyte / Macrophage Colony-Stimulating Factor (GM-CSF) and Interleukin-4. American Journal of Reproductive Immunology, 49: 230–238. doi: 10.1034/j.1600-0897.2003.01200.x
- Issue published online: 25 MAR 2003
- Article first published online: 25 MAR 2003
- Submitted July 7, 2002; revised September 9, 2002; accepted September 17, 2002.
- dendritic cell;
OBJECTIVE: We investigated how melanoma cells and membrane-bound granulocyte/macrophage colony stimulating factor (mbGM-CSF) melanoma cell lines affect the differentiation of dendritic cells (DC) from CD14+ monocytes.
METHOD OF STUDY: The malignant melanoma cell lines (Conley and Jorp) and mbGM-CSF-positive cell lines (Conley B-F8 and Jorp C-E6) were cultured and cell-free supernatants were collected from each cell line cultures to assess the GM-CSF level. Adherent monocytes were cocultured for 6–7 days with irradiated mbGM-CSF and wild type melanoma cells (50 Gy) at each 1 : 1 and 0.1 : 1 ratio in six-well culture plates in ex vivo culture medium containing interleukin (IL)-4. Immunophenotyping was performed by triple color immunofluorescence staining with flow cytometry analysis.
RESULTS: GM-CSF was detected at low levels in the culture supernatants of Conley B-F8 (0.48 ng/106 cells/24 hr), whereas there was no detectable GM-CSF in that of Conley melanoma cell line. Monocytes cultured with GM-CSF/IL-4 generated the expression of high levels of HLA DR, CD1a and CD86, while the expression of CD14 and CD83 were in low amounts. However, the addition of GM-CSF to these cultures resulted in an increased expression of these markers and decreased that of CD14. Monocytes cocultured with Jorp C-E6 illustrated similar expression pattern of CD1a, HLA DR and CD14 in the presence or absence of GM-CSF as Conley B-F8 melanoma cell line. Monocytes cocultured with Conley B-F8 melanoma cells at 1 : 1 and 0.1 : 1 ratio showed no significant difference in expression of CD1a, CD14 and CD83 between the two ratios.
CONCLUSION: Our results illustrate the feasibility to generate monocyte-derived DC from coculture with melanoma tumor cells in the presence of GM-CSF and IL-4. However, mbGM-CSF tumor cells did not significantly enhance the DC differentiation through juxtacrine stimulation unless soluble GM-CSF was added in the culture media.