Use of HLA-B27 tetramers to identify low-frequency antigen-specific T cells in Chlamydia-triggered reactive arthritis
Use of HLA-B27 tetramers to identify low-frequency antigen-specific T cells in Chlamydia-triggered reactive arthritis
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DOI：
10.1186/ar1221
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发表时间：
2004-01-01
影响因子：
4.9
通讯作者：
Sieper, J
中科院分区：
文献类型：
--
作者：
Appel, H;Kuon, W;Sieper, J
利用HLA-B27/肽四聚体复合物研究HLA-B27（+）相关疾病中肽特异性CD8(+) T细胞的报道很少。为了建立HLA-B27四聚体，我们首先利用病毒表位比较了HLA-B27四聚体与HLA-A2四聚体的功能。负载Epstein Barr病毒免疫优势肽的HLA-B27和HLA-A2四聚体的产量相当，两种分子都检测到抗原特异性CD8(+) T细胞。应用HLA-B27四聚体在HLA-B27相关疾病中的应用，研究了滑膜液和外周血中最近发现的9种衣原体衍生肽，以检测9例衣原体引发的反应性关节炎（CtReA）患者的CD8(+) T细胞对沙眼衣原体抗原的反应。6例HLA-B27(+) Ct-ReA患者中有4例滑膜T细胞与至少一种HLA-B27/衣原体肽四聚体特异性结合。HLA-B27/衣原体肽195四聚体与6例患者中的3例滑膜T细胞结合，HLA-B27/衣原体肽133四聚体与2例患者的滑膜T细胞结合。然而，这些细胞的频率很低（0.02-0.09%）。此外，我们展示了两种生成hla - b27限制性T细胞系的方法。首先，使用HLA-B27四聚体和磁珠对抗原特异性CD8(+) T细胞进行分选。其次，利用衣原体感染的树突状细胞体外刺激CD8(+) T细胞。利用负载EBV肽的HLA-B27四聚体进行体外磁分选，可获得高纯度的CD8 T细胞系。衣原体感染的树突状细胞体外刺激CD8(+) T细胞，可增加衣原体特异性、HLA-B27四聚体结合CD8(+) T细胞的频率。我们得出结论，HLA-B27四聚体是检测和扩增HLA-B27限制性CD8(+) T细胞的有用工具。在患有Ct-ReA的HLA-B27（+）患者的滑液中发现了一种或三种衣原体衍生肽中的一种或多种特异性T细胞。这些细胞可以在体外扩增，表明它们具有免疫功能。
Reports of the use of HLA-B27/peptide tetrameric complexes to study peptide-specific CD8(+) T cells in HLA-B27(+)-related diseases are rare. To establish HLA-B27 tetramers we first compared the function of HLA-B27 tetramers with HLA-A2 tetramers by using viral epitopes. HLA-B27 and HLA-A2 tetramers loaded with immunodominant peptides from Epstein Barr virus were generated with comparable yields and both molecules detected antigen-specific CD8(+) T cells. The application of HLA-B27 tetramers in HLA-B27-related diseases was performed with nine recently described Chlamydia-derived peptides in synovial fluid and peripheral blood, to examine the CD8(+) T cell response against Chlamydia trachomatis antigens in nine patients with Chlamydia-triggered reactive arthritis (CtReA). Four of six HLA-B27(+) Ct-ReA patients had specific synovial T cell binding to at least one HLA-B27/Chlamydia peptide tetramer. The HLA-B27/Chlamydia peptide 195 tetramer bound to synovial T cells from three of six patients and HLA-B27/Chlamydia peptide 133 tetramer to synovial T cells from two patients. However, the frequency of these cells was low (0.02-0.09%). Moreover, we demonstrate two methods to generate HLA-B27-restricted T cell lines. First, HLA-B27 tetramers and magnetic beads were used to sort antigen-specific CD8(+) T cells. Second, Chlamydia-infected dendritic cells were used to stimulate CD8(+) T cells ex vivo. Highly pure CD8 T cell lines could be generated ex vivo by magnetic sorting by using HLA-B27 tetramers loaded with an EBV peptide. The frequency of Chlamydia-specific, HLA-B27 tetramer-binding CD8(+) T cells could be increased by stimulating CD8(+) T cells ex vivo with Chlamydia-infected dendritic cells. We conclude that HLA-B27 tetramers are a useful tool for the detection and expansion of HLA-B27-restricted CD8(+) T cells. T cells specific for one or more of three Chlamydia-derived peptides were found at low frequency in synovial fluid from HLA-B27(+) patients with Ct-ReA. These cells can be expanded ex vivo, suggesting that they are immunologically functional.