Sánchez-Monge R, Blanco C, Perales A D, Collada C, Carrillo T, Aragoncillo C, Salcedo G
Unidad de Bioquímica, Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos, Madrid, Spain.
J Allergy Clin Immunol. 2000 Jul;106(1 Pt 1):190-5. doi: 10.1067/mai.2000.107599.
Class I chitinases have been identified as the major panallergens in fruits associated with the latex-fruit syndrome, such as avocado, banana, and chestnut. However, other plant foods containing these enzymes have not been related to this syndrome.
We sought out class I chitinases in the green bean, a legume that is known to express chitinases but is not associated with latex allergy, and examined whether the content or allergenic activity of chitinases can be modified by physical or chemical treatments.
IgE-binding proteins in untreated bean samples, as well as in ethylene- and heat-treated samples, were detected by using a pool of sera from patients with latex-fruit allergy. Putative allergens were purified by cation-exchange chromatography and characterized by N-terminal sequencing, enzymatic activity assays, immunodetection with sera and antichitinase antibodies, and immunoblot inhibition tests. Skin prick tests with untreated and heated purified allergens were also carried out.
An IgE-binding protein of 32 kd that was also recognized by antichitinase antibodies was detected in green bean extracts. This reactive component was strongly induced by ethylene treatment. The protein, designated PvChI, was identified as a class I chitinase closely related to the major avocado allergen Prs a 1. Immunoblot inhibition assays demonstrated cross-reactivity between both allergens. Purified PvChI induced positive skin prick test responses in 7 of 8 patients with latex-fruit allergy. Heat treatment of both Prs a 1 and PvChI produced a full loss of their allergenic capacities both in vitro and in vivo. No IgE-binding component was detected in the white mature bean in which the main isolated 32-kd protein corresponded to a nonreactive phytohemagglutinin.
Ethylene treatment induces the expression of plant class I chitinases. The allergenic activity of plant class I chitinases seems to be lost by heating. This fact could explain why plant foods containing these putative allergens that are consumed after cooking are not usually associated with the latex-fruit syndrome.
I类几丁质酶已被确定为与乳胶-水果综合征相关的水果中的主要泛过敏原,如鳄梨、香蕉和栗子。然而,其他含有这些酶的植物性食物与该综合征并无关联。
我们在绿豆中寻找I类几丁质酶,绿豆是一种已知能表达几丁质酶但与乳胶过敏无关的豆类,并研究几丁质酶的含量或致敏活性是否能通过物理或化学处理而改变。
使用乳胶-水果过敏患者的混合血清检测未处理的豆类样品以及经乙烯和热处理的样品中的IgE结合蛋白。通过阳离子交换色谱法纯化假定的过敏原,并通过N端测序、酶活性测定、用血清和抗几丁质酶抗体进行免疫检测以及免疫印迹抑制试验对其进行表征。还对未处理和加热的纯化过敏原进行了皮肤点刺试验。
在绿豆提取物中检测到一种32kd的IgE结合蛋白,该蛋白也能被抗几丁质酶抗体识别。乙烯处理强烈诱导了这种反应性成分。该蛋白被命名为PvChI,被鉴定为一种与主要鳄梨过敏原Prs a 1密切相关的I类几丁质酶。免疫印迹抑制试验证明了这两种过敏原之间的交叉反应性。纯化的PvChI在8名乳胶-水果过敏患者中的7名中诱导了阳性皮肤点刺试验反应。对Prs a 1和PvChI进行热处理后,它们在体外和体内的致敏能力完全丧失。在白色成熟豆中未检测到IgE结合成分,其中主要分离出的32kd蛋白对应于一种无反应性的植物血凝素。
乙烯处理可诱导植物I类几丁质酶的表达。植物I类几丁质酶的致敏活性似乎因加热而丧失。这一事实可以解释为什么烹饪后食用的含有这些假定过敏原的植物性食物通常与乳胶-水果综合征无关。