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Frequency of clonally expanded T cells evaluated by PCR from a single cell.
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2
Direct analysis of viral-specific CD8+ T cells with soluble HLA-A2/Tax11-19 tetramer complexes in patients with human T cell lymphotropic virus-associated myelopathy.用人T细胞嗜淋巴病毒相关脊髓病患者的可溶性HLA - A2/Tax11 - 19四聚体复合物直接分析病毒特异性CD8 + T细胞
J Immunol. 1999 Feb 1;162(3):1765-71.
3
Detailed chromosomal and molecular genetic analysis of single cells by whole genome amplification and comparative genomic hybridisation.通过全基因组扩增和比较基因组杂交对单细胞进行详细的染色体和分子遗传学分析。
Nucleic Acids Res. 1999 Feb 15;27(4):1214-8. doi: 10.1093/nar/27.4.1214.
4
Highly efficient recovery of functional single-chain Fv fragments from inclusion bodies overexpressed in Escherichia coli by controlled introduction of oxidizing reagent--application to a human single-chain Fv fragment.通过控制引入氧化试剂从大肠杆菌中过表达的包涵体中高效回收功能性单链Fv片段——应用于一种人单链Fv片段
J Immunol Methods. 1998 Oct 1;219(1-2):119-29. doi: 10.1016/s0022-1759(98)00127-6.
5
Induction and exhaustion of lymphocytic choriomeningitis virus-specific cytotoxic T lymphocytes visualized using soluble tetrameric major histocompatibility complex class I-peptide complexes.利用可溶性四聚体主要组织相容性复合体I类-肽复合物观察淋巴细胞性脉络丛脑膜炎病毒特异性细胞毒性T淋巴细胞的诱导和耗竭。
J Exp Med. 1998 May 4;187(9):1383-93. doi: 10.1084/jem.187.9.1383.
6
Clonotypic analysis of T cells in patients with autoimmune and viral hepatitis.自身免疫性肝炎和病毒性肝炎患者T细胞的克隆型分析。
Hepatology. 1997 May;25(5):1070-6. doi: 10.1002/hep.510250504.
7
Correlation of clonal T cell expansion with disease activity in systemic lupus erythematosus.系统性红斑狼疮中克隆性T细胞扩增与疾病活动的相关性
Int Immunol. 1997 Apr;9(4):547-54. doi: 10.1093/intimm/9.4.547.
8
A three-domain T cell receptor is biologically active and specifically stains cell surface MHC/peptide complexes.一种三结构域T细胞受体具有生物活性，并能特异性地标记细胞表面的MHC/肽复合物。
J Immunol. 1997 Mar 1;158(5):2218-27.
9
Characterization of T cell receptor single-chain Fv fragments secreted by myeloma cells.骨髓瘤细胞分泌的T细胞受体单链Fv片段的特性分析
Eur J Immunol. 1996 Oct;26(10):2410-6. doi: 10.1002/eji.1830261022.
10
Mammalian expression of single chain variable region fragments dimerized by Fc regions.由Fc区域二聚化的单链可变区片段的哺乳动物表达。
Mol Biol Rep. 1995;21(3):141-6. doi: 10.1007/BF00997236.

在不建立T细胞克隆的情况下，从单个T细胞中对T细胞受体α和β基因进行配对克隆。

Paired cloning of the T cell receptor alpha and beta genes from a single T cell without the establishment of a T cell clone.

作者信息

Kurokawa M, Tong J, Matsui T, Masuko-Hongo K, Yabe T, Nishioka K, Yamamoto K, Kato T

机构信息

Rheumatology, Immunology and Genetic Program, Institute of Medical Science, St Marianna University School of Medicine, Kawasaki, Japan.

出版信息

Clin Exp Immunol. 2001 Feb;123(2):340-5. doi: 10.1046/j.1365-2249.2001.01437.x.

DOI:10.1046/j.1365-2249.2001.01437.x
PMID:11207667
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1905975/
Abstract

T cell receptors, which recognize antigen peptides on MHC molecules, are essential probes for the analysis of T cell antigen specificity. The identification of paired T cell receptor (TCR) chains, alpha/beta or gamma/delta, usually requires the establishment of T cell clones, which is not always available. In this study, we tried, as an alternative method, the paired cloning of TCR alpha/beta genes directly from a single T cell. T cells were sorted as a single cell from which RNA was extracted. Then, TCR alpha/beta CDR3 regions were amplified from the single cell-derived cDNA by reverse transcriptase-polymerase chain reaction to determine their sequences. We successfully identified pairs of TCR alpha/beta genes, and reconstructed the TCR molecule by a bacterial expression system. This strategy makes it possible to obtain recombinant TCR molecules from a single T cell without cellular cloning and promotes the investigation of T cell antigen specificity.

摘要

T细胞受体可识别MHC分子上的抗原肽，是分析T细胞抗原特异性的重要探针。鉴定配对的T细胞受体（TCR）链，即α/β或γ/δ，通常需要建立T细胞克隆，但这并非总是可行。在本研究中，作为一种替代方法，我们尝试直接从单个T细胞中对TCRα/β基因进行配对克隆。将T细胞作为单个细胞分选出来，从中提取RNA。然后，通过逆转录-聚合酶链反应从单细胞来源的cDNA中扩增TCRα/β CDR3区域，以确定其序列。我们成功鉴定出TCRα/β基因对，并通过细菌表达系统重建了TCR分子。该策略使得无需细胞克隆即可从单个T细胞获得重组TCR分子，并促进了对T细胞抗原特异性的研究。