Molecular features of the complementarity determining region 3 motif of the T cell population and subsets in the blood of patients with chronic severe hepatitis B.
Molecular features of the complementarity determining region 3 motif of the T cell population and subsets in the blood of patients with chronic severe hepatitis B.
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DOI：
10.1186/1479-5876-9-210
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发表时间：
2011-12-08
影响因子：
7.4
通讯作者：
Li L
中科院分区：
文献类型：
--
作者：
Yang J;He J;Lu H;Wei L;Li S;Wang B;Diao H;Li L
T细胞受体（TCR）反映T细胞的状态和功能。我们之前开发了一种基因熔解谱（GMSP）检测方法，该方法通过使用定量实时逆转录PCR（qRT-PCR）和DNA熔解曲线分析快速检测HBV患者T细胞受体β可变基因（TCRBV）的克隆扩增。然而，慢性重型肝炎B（CSH B）患者外周血单个核细胞（PBMCs）和CD 8+、CD 8-细胞亚群中TCRBV的分子特征尚不清楚。使用密度梯度离心和磁激活细胞分选（MACS）分离人PBMC并分选成CD 8+和CD 8-细胞亚群。使用GMSP分析确定TCRBV CDR 3基序的分子特征;当GMSP谱显示单峰（指示单克隆群体）时，对TCRBV家族进行克隆和测序。通过GMSP分析评估，CD 8+细胞亚群中的偏斜TCRBV的数量显著高于CD 8-细胞亚群。TCRBV 11和BV 7在PBMC和CD 8+、CD 8-亚群中的表达频率高于TCRBV家族其他成员。CSHB患者PBMC中TCRBV 22、BV 18和BV 11 CDR 3也存在相对保守的氨基酸基序。CSHB患者外周血单个核细胞和CD 8+、CD 8-细胞亚群中TCRBV CDR 3的分子特征存在明显差异。分析CD 8+亚群中的TCRBV表达在评估循环T细胞的状态和功能方面更准确。TCRBV 11、BV 7和相对保守的CDR 3氨基酸基序的表达也有助于预测和治疗CSHB患者。
T cell receptor (TCR) reflects the status and function of T cells. We previously developed a gene melting spectral pattern (GMSP) assay, which rapidly detects clonal expansion of the T cell receptor β variable gene (TCRBV) in patients with HBV by using quantitative real-time reverse transcription PCR (qRT-PCR) with DNA melting curve analysis. However, the molecular profiles of TCRBV in peripheral blood mononuclear cells (PBMCs) and CD8+, CD8- cell subsets from chronic severe hepatitis B (CSHB) patients have not been well described. Human PBMCs were separated and sorted into CD8+ and CD8- cell subsets using density gradient centrifugation and magnetic activated cell sorting (MACS). The molecular features of the TCRBV CDR3 motif were determined using GMSP analysis; the TCRBV families were cloned and sequenced when the GMSP profile showed a single-peak, indicative of a monoclonal population. The number of skewed TCRBV in the CD8+ cell subset was significantly higher than that of the CD8- cell subset as assessed by GMSP analysis. The TCRBV11 and BV7 were expressed more frequently than other members of TCRBV family in PBMCs and CD8+, CD8- subsets. Also the relatively conserved amino acid motifs were detected in the TCRBV22, BV18 and BV11 CDR3 in PBMCs among patients with CSHB. The molecular features of the TCRBV CDR3 were markedly different among PBMCs and CD8+, CD8- cell subsets derived from CSHB patients. Analysis of the TCRBV expression in the CD8+ subset was more accurate in assessing the status and function of circulating T cells. The expression of TCRBV11, BV7 and the relatively conserved CDR3 amino acid motifs could also help to predict and treat patients with CSHB.
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影响因子：
56.9
作者：
Morgan, Richard A.;Dudley, Mark E.;Rosenberg, Steven A.
通讯作者：
Rosenberg, Steven A.
影响因子：
15.3
作者：
Das, Abhishek;Hoare, Matthew;Davies, Nathan;Lopes, A. Ross;Dunn, Claire;Kennedy, Patrick T. F.;Alexander, Graeme;Finney, Helene;Lawson, Alistair;Plunkett, Fiona J.;Bertoletti, Antonio;Akbar, Arne N.;Maini, Mala K.
通讯作者：
Maini, Mala K.
影响因子：
2.6
作者：
Du, Jin-Wei;Gu, Jiang-Ying;Zhu, Ping
通讯作者：
Zhu, Ping
影响因子：
24.5
作者：
Imamura, T;Yokosuka, O;Saisho, H
通讯作者：
Saisho, H
影响因子：
4.4
作者：
Melenhorst, J. Joseph;Lay, Matthew D. H.;Barrett, A. John
通讯作者：
Barrett, A. John