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T-cell epitopes of the major peach allergen, Pru p 3: Identification and differential T-cell response of peach-allergic and non-allergic subjects

机译:主要桃子变应原的T细胞表位,Pru p 3:桃子变态反应和非变态反应对象的鉴定和差异化T细胞反应

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摘要

Lipid transfer proteins (LTPs), particularly peach Pru p 3, are the most relevant plant food allergens in the South of Europe, and, therefore, their allergic properties have been extensively studied. However, neither T-cell epitopes nor their effect on the patients’ T-cell response has been investigated in any member of the LTP panallergen family. The objective of the present study was to map the major T-cell epitopes of Pru p 3, as well as to evaluate their induced T-cell response in peach-allergic versus control subjects. Thus, peripheral blood mononuclear cells (PBMCs) from 18 peach-allergic patients and Pru p 3-specific T-cell lines (TCLs) from 9 of them were cultured with Pru p 3 and with a panel of 17 derived peptides (10-mer overlapping in 5 amino acids representing the full sequence of Pru p 3). Proliferation in 5-day assays was carried out via tritiated-thymidine incorporation, while IL4 and IFNγ production was assessed via sandwich enzyme-linked immunosorbent tests (ELISA) of TCL culture supernatants. The results were compared to those obtained from 10 non-peach allergic control volunteers. Two consecutive peptides showed the highest activation capacity. About 74% of PBMCs and TCLs recognized them, forming a single T-epitope: Pru p 365–80. Additionally, other specific T-cell epitopes were observed. Pru p 325–35 was detected by more than 60% of TCLs from peach-allergic patients, and Pru p 345–55 only activated PBMCs from control subjects. Interestingly, TCLs from patients were associated with a Th2-type, whereas control TCLs presented a Th1-type cytokine response. The major immunogenic T-cell epitope identified in Pru p 3, Pru p 365–80, is a good candidate to develop new vaccines for hypersensitivity reactions associated with LTP allergens from Rosaceae fruits.
机译:脂质转移蛋白(LTPs),特别是桃Pru p 3,是欧洲南部最相关的植物性食物过敏原,因此,已经广泛研究了它们的过敏特性。但是,在LTP全变应原家族的任何成员中,都没有研究过T细胞表位及其对患者T细胞反应的影响。本研究的目的是绘制Pru p 3的主要T细胞表位,并评估桃子过敏与对照对象中诱导的T细胞反应。因此,将来自18位桃子过敏患者的外周血单核细胞(PBMC)和来自其中9位患者​​的Pru p 3特异性T细胞系(TCL)与Pru p 3和一组17种衍生肽(10-mer 5个氨基酸重叠表示Pru p 3的完整序列)。通过掺入tri代胸腺嘧啶核苷进行5天试验的增殖,同时通过TCL培养物上清液的夹心酶联免疫吸附试验(ELISA)评估IL4和IFNγ的产生。将结果与从10名非桃类过敏对照志愿者获得的结果进行比较。两个连续的肽显示出最高的激活能力。大约74%的PBMC和TCL识别出它们,形成了一个T型表位:Pru p 365–80。另外,观察到其他特定的T细胞表位。桃过敏患者中超过60%的TCL检测到Pru p 325-35,而Pru p 345-55仅激活了对照组的PBMC。有趣的是,来自患者的TCL与Th2型相关,而对照TCL呈现Th1型细胞因子反应。 Pru p 3(Pru p 365-80)中鉴定出的主要免疫原性T细胞表位是开发与酒渣鼻果实LTP过敏原相关的超敏反应新疫苗的良好候选者。

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