Simultaneous detection of circulating autoreactive CD8+ T-cells specific for different islet cell-associated epitopes using combinatorial MHC multimers.
Simultaneous detection of circulating autoreactive CD8+ T-cells specific for different islet cell-associated epitopes using combinatorial MHC multimers.
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作者：
Velthuis JH;Unger WW;Abreu JR;Duinkerken G;Franken K;Peakman M;Bakker AH;Reker-Hadrup S;Keymeulen B;Drijfhout JW;Schumacher TN;Roep BO
1 型糖尿病是由 T 细胞选择性破坏胰腺中产生胰岛素的 β 细胞引起的。在此过程中，胰岛表位特异性 CD8+ T 细胞发挥着关键作用。因此，监测多个胰岛特异性 CD8+ T 细胞可能对测量疾病活动、进展和干预有价值。然而，传统的检测技术（ELISPOT 和 HLA 四聚体）需要许多细胞并且相对不敏感。在这里，我们使用组合量子点主要组织相容性复合物多聚体技术来同时监测 HLA-A2 限制性胰岛素 B10-18、前胰岛素原 (PPI)15-24、胰岛抗原 (IA)-2797-805、GAD65114-123、胰岛特异性葡萄糖-6-磷酸酶催化的存在。 新近发病的糖尿病患者、其兄弟姐妹、健康对照受试者和胰岛细胞移植受者中的亚基相关蛋白 (IGRP)265–273 和前原胰岛淀粉样多肽 (ppIAPP)5–13 特异性 CD8+ T 细胞。使用该试剂盒，识别胰岛素 B10-18、IA-2797-805 和 IGRP265-273 的胰岛自身反应性 CD8+ T 细胞在新近发病的糖尿病患者中经常可检测到，但在健康对照受试者中很少检测到； PPI15-24 被证明是最敏感的表位。将“Diab-Q-kit”应用于胰岛细胞移植受者的样本，可以检测针对多个胰岛细胞衍生表位的自身反应性 T 细胞频率的变化，这些表位与疾病活动性相关并与临床结果相关。我们开发了一种试剂盒，可以同时检测对多个 HLA-A2 限制性 β 细胞表位有反应的 CD8+ T 细胞，只需少量血液，无需体外培养，适用于储存的血液样本。
Type 1 diabetes results from selective T-cell–mediated destruction of the insulin-producing β-cells in the pancreas. In this process, islet epitope–specific CD8+ T-cells play a pivotal role. Thus, monitoring of multiple islet–specific CD8+ T-cells may prove to be valuable for measuring disease activity, progression, and intervention. Yet, conventional detection techniques (ELISPOT and HLA tetramers) require many cells and are relatively insensitive. Here, we used a combinatorial quantum dot major histocompatibility complex multimer technique to simultaneously monitor the presence of HLA-A2 restricted insulin B10–18, prepro-insulin (PPI)15–24, islet antigen (IA)-2797–805, GAD65114–123, islet-specific glucose-6-phosphatase catalytic subunit–related protein (IGRP)265–273, and prepro islet amyloid polypeptide (ppIAPP)5–13–specific CD8+ T-cells in recent-onset diabetic patients, their siblings, healthy control subjects, and islet cell transplantation recipients. Using this kit, islet autoreactive CD8+ T-cells recognizing insulin B10–18, IA-2797–805, and IGRP265–273 were shown to be frequently detectable in recent-onset diabetic patients but rarely in healthy control subjects; PPI15–24 proved to be the most sensitive epitope. Applying the “Diab-Q-kit” to samples of islet cell transplantation recipients allowed detection of changes of autoreactive T-cell frequencies against multiple islet cell–derived epitopes that were associated with disease activity and correlated with clinical outcome. A kit was developed that allows simultaneous detection of CD8+ T-cells reactive to multiple HLA-A2–restricted β-cell epitopes requiring limited amounts of blood, without a need for in vitro culture, that is applicable on stored blood samples.
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DOI：
10.1073/pnas.0508621102
发表时间：
2005-12-20
影响因子：
11.1
作者：
Pinkse, GGM;Tysma, OHM;Roep, BO
通讯作者：
Roep, BO
影响因子：
32.4
作者：
Ernst, B;Lee, DS;Surh, CD
通讯作者：
Surh, CD
影响因子：
15.9
作者：
Skowera, Ania;Ellis, Richard J.;Peakman, Mark
通讯作者：
Peakman, Mark
影响因子：
56.9
作者：
Altman, JD;Moss, PAH;Davis, MM
通讯作者：
Davis, MM
自身免疫性糖尿病中针对谷氨酸脱羧酶特异的细胞毒性T细胞。
影响因子：
15.3
作者：
Panina-Bordignon, Paola;Lang, Rosmarie;Van Endert, Peter M.;Benazzi, Elena;Felix, Arthur M.;Pastore, Rocco M.;Spinas, Giatgen A.;Sinigaglia, Francesco
通讯作者：
Sinigaglia, Francesco