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Development of animal models for cutaneous eosinophilic inflammation and analysis of its regulatory mechanisms

皮肤嗜酸性炎症动物模型的建立及其调控机制分析

基本信息

批准号：
18591260
负责人：
TERUI Tadashi
金额：
$ 2.52万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591260/
关键词：
skin eosinophilic inflammation mast cell animal model

项目摘要

Recent studies suggest that mast cell derived-TNF-alpha plays important roles in the development of contact hypersensitivity induced by hapten. Recently, we revealed that Fc epsilon RI beta-chain controls production of proinflammatory cytokines including TNF-alpha from mast cells through function of three tyrosine residues (Y219/Y229/Y225) of its ITAM. In the present study, we investigated the biological functions of mast cells expressing wild-type or mutated β-chain ITAM in development of the contact hypersensitivity employing mast cell "knock-in" mice.We prepared mast cells harboring wild-type (YYY) or Fc epsilon RI beta-chain ITAM mutant (FFF) by employing a retrovirus-mediated gene transfer into BMMCs derived from Fc epsilon RI beta-chain -/- mice. Those mast cells were intradermally transferred into ears of mast cell deficient mice (W/Wv). The mice were pre-sensitized by application of 2% oxazolone to the shaved abdomen. On day 5, 1% oxazolone and vehicle were applied to both sides of right ear and left ear, respectively. Contact hypersensitivity was evaluated by ear thickness.Cytokine expression in the tissue was analyzed by Real-Time PCR.As a result, oxazolone failed to cause ear swelling in W/Wv and Fc epsilon RI beta-chain -/- mice. In addition, we showed that ear swelling is significantly reduced in the FFF-knock-in mice as compared with that of YYY- knock-in mice. Consistent with this result, infiltration of CD3^<+> T cells and eosinophils into the inflammation sites was severely reduced in FFF-knock-in mice. Furthermore, an experiment using real-time PCR demonstrated that expression of TNF-alpha mRNA is also decreased in the ear tissue of FFF-knock-in mice. Taken together, these results suggest that regulation of mast cell activation through Fc epsilon RI beta-chain is critical for development of contact hypersensitivity following cutaneous exposure of hapten.

最近的研究表明，肥大细胞源性TNF-α在半抗原诱导的接触性超敏反应中起重要作用。最近，我们发现Fc β RI β链通过其ITAM的三个酪氨酸残基（Y219/Y229/Y225）的功能控制促炎细胞因子（包括TNF-α）从肥大细胞的产生。在本研究中，我们研究了表达野生型或突变型β-链ITAM的肥大细胞在肥大细胞“敲入”小鼠的接触性超敏反应中的生物学功能，我们通过逆转录病毒介导的基因转移到来自Fc β-RI β-chain-/-小鼠的BMMC中，制备了携带野生型（YYY）或Fc β-RI β-chain ITAM突变体（FFF）的肥大细胞。将这些肥大细胞皮内转移到肥大细胞缺陷型小鼠（W/Wv）的耳中。通过将2%恶唑酮应用于剃毛的腹部使小鼠预致敏。在第5天，将1%恶唑酮和赋形剂分别应用于右耳和左耳的两侧。通过耳厚度评价接触性过敏反应，通过Real-Time PCR分析组织中细胞因子的表达。结果，恶唑酮未引起W/Wv和Fc ε RI β链-/-小鼠的耳肿胀。此外，我们表明，与YYY敲入小鼠相比，FFF敲入小鼠的耳肿胀显著减少。与该结果一致，在FFF敲入小鼠中，CD 3 ^<+> T细胞和嗜酸性粒细胞向炎症部位的浸润严重减少。此外，使用实时PCR的实验表明，在FFF敲入小鼠的耳组织中TNF-α mRNA的表达也降低。总之，这些结果表明，通过Fc β RI β链调节肥大细胞活化对于皮肤暴露于半抗原后发生接触性超敏反应至关重要。