| 研究実績の概要 |
T cell is a major player in the immune system, it can kill cancer cells when its unique T cell receptor (TCR; Each T cell has different TCR) specifically recognizes an antigen on the cancer cell, but must be activated inducing proliferation by antigen in advance. There are several types of immunotherapies can help the body to activate or generate the cancer specific T cells. However, these immunotherapies all need the antigen-TCR pair information, and it is difficult to find a specific TCR-antigen pair using the current established methods. The purpose here is to develop a novel high-throughput platform for TCR-antigen pair identification, which can identify both the cancer-related antigens and the specific TCR sequences for these antigens.
In this fiscal year, we have picked up the T cells by an established picking-up system. The TCR (T cell receptor) sequences were amplified from single T cells, and some of them were confirmed by Sanger sequencing. We used a T cell which has the known TCR sequence, and we confirmed that the amplified sequences are correct.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
(1) Cell picking-up.
We have built-up a custom cell picking-up system which is used for this project. We have picked up the T cells by an established picking-up system to the PCR tubes.
(2) TCR amplification
The TCR α and β chains were amplified by reverse transcription and PCR. Multiple conditions were screened to improve the amplification efficiency.
(3) TCR sequence identification
The amplified TCR cDNA were sequenced by Sangar sequencing. For control T cells, we compared the identified sequences with the known TCR sequence, which confirmed that the identified TCR sequence were correct.
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| 今後の研究の推進方策 |
In this fiscal year (2019.4.1~2019.8.31), I will discriminate different cases of T cell activity by imaging, and pick up the different T cells.
(1) find the different cases of T cell activity using imaging
(2) pick up the different T cell to PCR tubes.
(3) amplify the TCR from picked up T cells.
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