論文 - 詳細
| RRC ID | 38208 |
|---|---|
| 著者 | Arakaki A, Ooya K, Akiyama Y, Hosokawa M, Komiyama M, Iizuka A, Yamaguchi K, Matsunaga T. |
| タイトル | TCR-beta repertoire analysis of antigen-specific single T cells using a high-density microcavity array. |
| ジャーナル | Biotechnol Bioeng |
| Abstract |
The antigen specificity of cytotoxic T cells, provided by T-cell receptors (TCRs), plays a central role in human autoimmune diseases, infection, and cancer. As the TCR repertoire is unique in individual cytotoxic T cells, a strategy to analyze its gene rearrangement at the single-cell level is required. In this study, we applied a high-density microcavity array enabling target cell screening of several thousands of single cells for identification of functional TCR-beta gene repertoires specific to melanoma (gp100) and cytomegalovirus (CMV) antigens. T cells expressing TCRs with the ability to recognize fluorescent-labeled antigen peptide tetramers were isolated by using a micromanipulator under microscopy. Regularly arranged cells on the microcavity array eased detection and isolation of target single cells from a polyclonal T-cell population. The isolated single cells were then directly utilized for RT-PCR. By sequencing the amplified PCR products, antigen-specific TCR-beta repertoires for gp100 and human cytomegalovirus antigens were successfully identified at the single-cell level. This simple, accurate, and cost-effective technique for single-cell analysis has further potential as a valuable and widely applicable tool for studies on gene screening and expression analyses of various kinds of cells. |
| 巻・号 | 106(2) |
| ページ | 311-8 |
| 公開日 | 2010-6-1 |
| DOI | 10.1002/bit.22663 |
| PMID | 20073087 |
| MeSH | Cell Separation / instrumentation* Equipment Design Equipment Failure Analysis Oligonucleotide Array Sequence Analysis / instrumentation* Receptors, Antigen, T-Cell / genetics* Reverse Transcriptase Polymerase Chain Reaction / instrumentation* |
| IF | 4.002 |
| 引用数 | 10 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| ヒト・動物細胞 | |