2015 Fiscal Year Final Research Report
Immune response against sarcoma stem-like cell antigen.
| Project/Area Number |
25462344
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Sapporo Medical University |
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Principal Investigator |
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Keywords | sarcoma stem cell / CTL clone / immunotherapy |
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| 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.
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| Free Research Field |
骨軟部腫瘍学・腫瘍免疫学・癌幹細胞
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