González Eva María, Villalba Mayte, Quiralte Joaquín, Batanero Eva, Roncal Fernando, Albar Juan Pablo, Rodríguez Rosalía
Departamento Bioquímica y Biología Molecular, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Mol Immunol. 2006 Feb;43(6):570-8. doi: 10.1016/j.molimm.2005.04.015. Epub 2005 Jun 6.
Ole e 1 is a major allergen from olive pollen with an IgE-binding frequency around 80% among allergic population. Its diagnostic value has been demonstrated, and cross-reactive allergens have been found in ash, lilac and privet. We sought to determine IgE- and IgG-binding regions of Ole e 1. Ole e 1-specific polyclonal antiserum and sera from patients allergic to olive pollen were used to analyze IgG and IgE epitopes, respectively. Short overlapping synthetic peptides covering the complete sequence of Ole e 1 and point mutants of these peptides bound to membranes, as well as long recombinant peptides fused to GST were used in dot blot immunostaining and ELISA. Skin prick tests were performed on 14 allergic patients to assay the response in vivo to the recombinant fusion peptides. Residues at positions 8-11, 29, 32, 33, 55-59, 70, 107-110, 112, 120, 123, 141 of Ole e 1 sequence were found to be antigenically relevant in the IgG-binding. Although amino acids K137, L138, G139, Y141 and P142 were involved in the IgE-recognition of a pool of sera from allergic individuals, the response to the IgEs seemed to be preferentially conformational. IgE-binding capability of recombinant GST-fused peptide T114-M145 was demonstrated by in vivo (prick test) and in vitro (ELISA) experiments. Major IgG and IgE-binding regions of Ole e 1 have been identified being the C-terminal an immunodominant region. These data could help to design hypoallergenic forms of the allergen.
油橄榄1(Ole e 1)是来自油橄榄花粉的一种主要变应原,在过敏人群中IgE结合频率约为80%。其诊断价值已得到证实,并且在白蜡树、丁香和女贞中发现了交叉反应性变应原。我们试图确定Ole e 1的IgE和IgG结合区域。分别使用Ole e 1特异性多克隆抗血清和对油橄榄花粉过敏患者的血清来分析IgG和IgE表位。覆盖Ole e 1完整序列的短重叠合成肽及其点突变体与膜结合,以及与谷胱甘肽S-转移酶(GST)融合的长重组肽用于斑点印迹免疫染色和酶联免疫吸附测定(ELISA)。对14名过敏患者进行皮肤点刺试验,以检测体内对重组融合肽的反应。发现Ole e 1序列中第8 - 11、29、32、33、55 - 59、70、107 - 110、112、120、123、141位的残基在IgG结合中具有抗原相关性。尽管氨基酸K137、L138、G139、Y141和P142参与了对过敏个体血清池的IgE识别,但对IgE的反应似乎优先是构象性的。通过体内(点刺试验)和体外(ELISA)实验证实了重组GST融合肽T114 - M145的IgE结合能力。已确定Ole e 1的主要IgG和IgE结合区域是C末端的一个免疫显性区域。这些数据有助于设计该变应原的低变应原性形式。