Hepatitis B surface antigen- and tetanus toxoid-specific clonal expansion of CD4+ cells in vitro determined by TCRBV CDR3 length and nucleotide sequence.
Hepatitis B surface antigen- and tetanus toxoid-specific clonal expansion of CD4+ cells in vitro determined by TCRBV CDR3 length and nucleotide sequence.
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通过 TCRBV CDR3 长度和核苷酸序列确定 CD4 细胞体外乙型肝炎表面抗原和破伤风类毒素特异性克隆扩增。
DOI：
10.1038/sj.gene.6363729
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Alper,CA
中科院分区：
文献类型：
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作者：
Uko,GP;Fraser,PA;Awdeh,ZL;Fici,DA;Crawford,KD;Larsen,CE;Alper,CA
我们证明了原代人TCRBV特异性CD 4+细胞在体外对B肝炎表面抗原（HBsAg）和破伤风类毒素（TT）的活化，而不使用细胞系，克隆或添加的细胞因子。通过多重PCR分析和光谱分析，抗原激活的细胞在特定和有限的TCRBV家族中表现出克隆性T细胞受体扩增。扩增的CD 4 + T细胞为CD 45 RO+。4例无关HBsAg应答者中有3例在TCRBV 16家族内显示CD 4+扩增。在所有三个供体中，应答主要包括单个CDR 3序列，其中一个供体完全是单克隆的。然而，应答者之间的CDR 3长度和序列不同。HBsAg诱导的TCRBV 16细胞克隆性与TT诱导的TCRBV 16细胞克隆性在序列、核苷酸添加和多样性（BD）或连接（BJ）元件使用方面具有特异性、可重复性和不同性。因此，对于第一次，我们显示了单克隆或寡克隆扩增的原代人CD 4+外周血单核细胞（PBMC）在体外响应于标称蛋白抗原，而无需操纵利用外源性IL-2。诱导对HBsAg的单克隆/寡克隆应答的能力现在允许进行基序鉴定研究，以确定T细胞在对HBsAg疫苗无应答中的作用。
We demonstrate activation of primary human TCRBV-specific CD4+ cells in vitro towards hepatitis B surface antigen (HBsAg) and tetanus toxoid (TT) without the use of cell lines, clones or added cytokines. By multiplex PCR analysis and spectratyping, antigen-activated cells exhibited clonal T cell receptor expansion within specific and limited TCRBV families. The expanded CD4+ T cells were CD45RO+. Three of four unrelated HBsAg responders showed CD4+ expansion within the TCRBV16 family. The response comprised predominantly single CDR3 sequences in all three donors and was completely monoclonal in one of them. However, the CDR3 lengths and sequences differed among the responders. Clonality induced by HBsAg in TCRBV16 was specific, reproducible and distinct from that induced by TT in terms of sequence, nucleotide addition and diversity (BD) or junctional (BJ) element usage. Thus, for the first time, we show monoclonal or oligoclonal expansion of primary human CD4+ peripheral blood mononuclear cells (PBMC) in vitro in response to nominal protein antigen without manipulations utilizing exogenous IL-2. The ability to induce monoclonal/oligoclonal responses to HBsAg now permits motif identification studies for determining the T cell role in non-responsiveness to the HBsAg vaccine.