| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
|
|---|
| Outline of Final Research Achievements |
We isolated sarcoma stem-like cells (SSC) from the malignant fibrous histiocytoma cell line, MFH2003. After autologous PBMC was stimulated with SSC, we established CTL clone Tc4c-6 recognizing SSC in the context of HLA class I. Next, we isolated cDNA of T cell receptor (TCR) alpha (A) & beta (B) chains. Two TCRAs were isolated (A5 and A7). Each cDNA of TCRA5B and TCRA7B was transfected into 58a-b- cells lacking TCR (58A5B and 58A7B). 58A7B cells could specifically recognize SSCs, however, the response was weak. Next, we transfected TCRA7B into the Jurkat cell line expressing NFAT-luciferase (Jrt/MA). The resultant cell line (J-A7B) recognized autologous SSCs but not non-SSCs. Moreover, J-A7B also recognized allogeneic osteosarcoma cell line KIKU in the context of HLA class I. Now we are trying to determine the HLA class I allele that presents the CTL epitope peptide derived from SSC antigen. These findings are useful for cDNA expression cloning of autologous CTL-defined SSC antigen.
|
