Isolation and characterization of native Cry j 3 from Japanese cedar (Cryptomeria japonica) pollen
Isolation and characterization of native Cry j 3 from Japanese cedar (Cryptomeria japonica) pollen
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DOI：
10.1111/j.1398-9995.2007.01331.x
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发表时间：
2007-05-01
期刊：
影响因子：
12.4
通讯作者：
Sakaguchi, M.
中科院分区：
文献类型：
--
作者：
Fujimura, T.;Futamura, N.;Sakaguchi, M.
背景：日本雪松花粉症是日本最常见的过敏性疾病。最近，日本雪松花粉过敏原Cry j 3被克隆为来自雪松(Juniperus Ashei)花粉的主要过敏原jun a 3的同源物。然而，天然的Cry j 3还没有被分离出来，也没有关于它的致敏活性的报道。本研究的目的是从雪松花粉中分离纯化天然Cry j 3，并对其免疫球蛋白E(IgE)结合能力进行评价。我们用免疫印迹分析和酶联免疫吸附试验检测了对雪松花粉过敏的患者的血清的IgE结合能力。此外，我们还检测了Cry j3诱导患者白细胞释放组胺的能力。我们从日本雪松花粉的cDNA文库中克隆了与纯化的Cry j 3相对应的基因。对雪松花粉过敏患者血清中Cry-j3的IgE结合率为27%(27/100)。CryJ3可诱导白细胞释放组胺。我们克隆了该基因并将其命名为Cry j 3.8。结论：Cryj3是雪松花粉症的致病变应原，在与口腔变态反应综合征的交叉反应中可能起重要作用。
Background: Japanese cedar (Cryptomeria japonica) pollinosis is the most prevalent allergy in Japan. Recently, the Japanese cedar pollen allergen Cry j 3 was cloned as a homologue of Jun a 3, which is a major allergen from mountain cedar (Juniperus ashei) pollen. However, native Cry j 3 has not been isolated and there are no reports on its allergenic activity. The aims of this study were to isolate native Cry j 3 and assess its immunoglobulin E (IgE)-binding capacity in patients with Japanese cedar pollinosis.Methods: Native Cry j 3 was purified from Japanese cedar pollen by multidimensional chromatography. We assessed the IgE-binding capacity using sera from patients allergic to Japanese cedar pollen by immunoblot analysis and ELISA. Moreover, we assayed the capacity of Cry j 3 to induce histamine release from the patients' leukocytes. We cloned cDNA corresponding to purified Cry j 3 from a cDNA library of Japanese cedar pollen.Results: We isolated native Cry j 3 as a 27-kDa protein. The IgE-binding frequency of Cry j 3 from the sera of patients allergic to Japanese cedar pollen was estimated as 27% (27/100) by ELISA. Cry j 3 induced the release of histamine from leukocytes. We cloned the cDNA and named it Cry j 3.8. Cry j 3.8 cDNA encoded 225 amino acids and had significant homology with thaumatin-like proteins.Conclusions: Cry j 3 is a causative allergen in Japanese cedar pollinosis and may play crucial roles in the cross-reactivity with oral allergy syndrome.