| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1999: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1998: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Research Abstract |
A monoclonal antibody (Mab JT-95) against thyroid carcinoma has been developed. A thyroid carcinoma cell line, designed SW1736 which express the sialyl fibronectin(FN) around cell surface as antigen of Mab JT-95, was incubated with Mab JT-95(100 μs/ml) and human lymphocytes at 37℃ in 72 hours. Another thyroid carcinoma cell line, TT, which does not express the sialyl fibronectin, was also incubated. Many cell lyses was observed in SW1736 after incubation. In contrast, a few or no cell lysis was seen in TT cell line.
We hae a hypothesis that the cell death might be induced by binding Mab JT-95 and FN from these results. To investigate this hypothesis,
1) the dose titration of MAb JT-95 (10,100,1000,10000 μs/ml) were made, and added to supernatant of SW1736 and TT, respectively. After 72hrs incubation at 37℃, colorimetric assay and ィイD13ィエD1T-Thymidine incorporate assay were performed. The dead cells increased parallel to dose depending of MAb JT-95 in SW1736. On the other hand, there is no change in TT cell line
2) Several peptides whichis compatible to a part of fibronectine were prepared and added to the SW1736 supernatant under the same concentration (100 μs/ml) of Mab JT-95 and incubated. From this experiment, a result revealed that the peptide of heparin binding domain which localizes near the C-terminal portion inhibited the cell death.
3) Heparin was also titrated and added to SW1736 with the same manner of MAb JT-95, and incubated. The dead cell increasing according to the concentration of heparin was seen.
4) SW1736 cells with Mab JT-95 incubation was lysised and extracted DNA to see the DNA laddering by gel electrophoresis. Tunel Method also performed. The cell death induced Mab JT-95 and heparin was apoptosis.
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