| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2002: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 2001: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2000: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1999: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Research Abstract |
Dendritic Cells are the most potent antigen presenting cells that initiate primary immune responses. The aims of this study are to analyze the composition of umbilical cord blood cells and to examine the characteristics of dendritic cells before and after culture. Umbilical cord blood cells were freshly determined the cell surface antigen, and then after culturing for 1, 2, 3, and 4 weeks with GM-CSF, IL-3 and SCF, every week they were determined the antigens. Method of flowcytometry was used to determine the number of dendritic cells and cell surface antigens before and after culture by using monoclonal antibody. Monoclonal antibodies included CD4, CD8, CD19, CD34, Cd83, CD1a, CD11c and CDw123. In the fresh umbilical cord blood, CD4^+ cells are 48.1%, CD8^+ 19.6, CD19^+ 7.5%, CD34^+ 2.9%, CD38^+ 73.5%, CD83^+ 1.5%, CD1a^+ 0.3% and CD11c^+ 12.2%. By the way, in fresh umbilical cord blood cells, total cells of CD83^+ were 1.9±1.1x10^5/ml, CD1a^+ 0.3±0.2x10^5/ml, CD11c^+ 5.9±2.5x10^5/ml and CDw123^+ 2.7±2.2x10^5/ml. On the other hand, after having cultured for 3 weeks, total cells of CD CD83^+ were 9.2±5.2x10^5/ml, CD1a^+ 9.8±9.3x10^5/ml, CD11c^+ 23.2±11.5x10^5/ml and CDw123^+ 19.6±10.1x10^5/ml.
As characteristic of umbilical cord blood after having cultured with cytokines of SCF, GM-CSF and IL-3, average population of lymphocytes decreased gradually, while average numbers of monocytes increased gradually in population. By culture with cytokine, dendritic cells increased up 3.1%-53.4%, and 6.9-28.2x10^5/ml in actual numbers. In comparison with fresh cord blood and their three-week cultured blood, CD83^+ active numbers were about 5.4 times, CD1a^+ 36.7 times, CD11c^+ 4.8 times and CDw123^+ 6.8 times. Umbilical cord blood cells cultured with cytokines differentiated into CD1a^+, CD83^+, CD11c^+ and CD83^+ dendritic cells. It seems these dendritic cells are very effective as antigen presenting cell and for cancer immunotherapy.
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