Raulf-Heimsoth M, Chen Z, Rihs H P, Kalbacher H, Liebers V, Baur X
Research Institute for Occupational Medicine (BGFA), Ruhr-University Bochum, Germany.
Clin Exp Allergy. 1998 Mar;28(3):339-48. doi: 10.1046/j.1365-2222.1998.00230.x.
The rubber elongation factor in Hevea rubber (Hev b 1) is one of the important latex-allergenic proteins inducing the production of specific IgE antibodies in latex-exposed subjects. In addition, Hev b 1 induces lymphocyte proliferation responses. A study about detailed T-cell epitope analysis of major latex allergens has not been published yet.
The aim of this study was to define the T-cell reactive regions on the latex allergen Hev b 1.
Nine overlapping peptides with 19 or 17 amino acid lengths representing the complete sequence of Hev b 1 were used for T-cell epitope mapping. Peripheral blood mononuclear cells (PBMCs) of latex-sensitized patients and healthy subjects were isolated and stimulated with the synthetic peptides to determine the specific proliferation response. The examined patients were sensitized to latex by occupational exposure (n=14) and suffered from rhinitis, conjunctivitis, contact urticaria and/or asthma. Two control groups of non-sensitized subjects were studied, one occupationally exposed to latex (n=4), the other one not exposed to latex (n=4).
Positive proliferation response induced by one or more peptides was detected in 10 of the latex-sensitized patients and in two of the latex-exposed non-sensitized subjects. Each patient exhibited an individual epitope reactivity pattern. The whole protein was found to be immunogenic, but the intensity of lymphocyte responses induced by the peptides was different. The majority of the patients' PBMCs (more than 66%) responded to the peptides 31-49 and 91-109. Using the peptide selection of Stille's algorithms, three epitopes were predicted covering region 38-74 and two others covering region 82-104; the two peptides 31-49 and 91-109 are part of these regions. A strong HLA-DR4Dw4 (DRB1*0401)-binding motif according to published DR4w4 binding motifs was predicted in the region of 102-110 including peptide 91-109 to which the majority of tested HLA-DR4-positive patients responded.
Our data indicate that the whole 14 kDa protein Hev b 1 is immunogenic having regions that demonstrate individual and varying PBMC stimulation. Experimental data of T-cell reactive regions based on PBMC-stimulation complement the information on T-cell epitope prediction. In addition, the Hev b 1 molecule contains a HLA-DR4Dw4 (DRB1*0401)-binding motif.
橡胶树橡胶中的橡胶延伸因子（Hev b 1）是重要的乳胶变应原性蛋白之一，可诱导接触乳胶的个体产生特异性IgE抗体。此外，Hev b 1可诱导淋巴细胞增殖反应。关于主要乳胶变应原详细T细胞表位分析的研究尚未见报道。
本研究旨在确定乳胶变应原Hev b 1上的T细胞反应区。
使用9个长度为19或17个氨基酸的重叠肽，它们代表Hev b 1的完整序列，用于T细胞表位定位。分离乳胶致敏患者和健康受试者的外周血单个核细胞（PBMC），并用合成肽刺激以确定特异性增殖反应。所检测的患者因职业暴露而对乳胶致敏（n = 14），患有鼻炎、结膜炎、接触性荨麻疹和/或哮喘。研究了两个未致敏受试者对照组，一组为职业性接触乳胶者（n = 4），另一组为未接触乳胶者（n = 4）。
在10例乳胶致敏患者和2例接触乳胶的未致敏受试者中检测到一种或多种肽诱导的阳性增殖反应。每位患者均表现出个体表位反应模式。发现整个蛋白具有免疫原性，但肽诱导的淋巴细胞反应强度不同。大多数患者的PBMC（超过66%）对肽31 - 49和91 - 109有反应。使用斯蒂勒算法的肽选择法，预测了3个表位覆盖38 - 74区域，另外2个覆盖82 - 104区域；肽31 - 49和91 - 109是这些区域的一部分。根据已发表的DR4w4结合基序，在102 - 110区域预测到一个强HLA - DR4Dw4（DRB1*0401）结合基序，包括大多数检测的HLA - DR4阳性患者有反应的肽91 - 109。
我们的数据表明，整个14 kDa蛋白Hev b 1具有免疫原性，其某些区域可显示个体性和不同的PBMC刺激。基于PBMC刺激的T细胞反应区实验数据补充了T细胞表位预测信息。此外，Hev b 1分子包含一个HLA - DR4Dw4（DRB1*0401）结合基序。
