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，肥大细胞得出的TNF-α在HAPEN引起的接触超敏反应中起着重要作用。最近，我们透露，FC Epsilon RIβ链控制促炎细胞因子的产生，包括来自肥大细胞的TNF-α通过其ITAM的三种酪氨酸残基（Y219/Y229/Y225）的功能。在本研究中，我们调查了表达野生型或突变的β-链ITAM的肥大细胞的生物学功能，从而开发了使用肥大细胞“敲入”鼠标的接触性超敏反应。我们准备了肥大细胞，我们准备了野生型（yyy）或FC Epsilon ri beta-beta-beta-ri beta-ri beta-ri betaim in nm rectem（fff fff）的肥大细胞，该细胞通过探索了fff（fff），该细胞通过使用bbm nmer（fff）转移了bmm betain retovir，该细胞通过逐步转移了bmm bet nmers（fff FFF），以循环转移了bmm contem（ FC Epsilon RIβ链 - / - 小鼠。这些肥大细胞在皮内转移到肥大细胞缺乏小鼠的耳朵中（W/WV）。小鼠通过在剃须腹部施加2％的沙发唑酮来敏感。在第5天，分别在右耳和左耳的两侧施加了1％的恶唑酮和媒介物。通过耳朵厚度评估了接触超敏反应。通过实时PCR分析组织中的循环反应。结果，恶唑酮未能导致W/WV和FC Epsilon RIβ链 - / - 小鼠的耳朵肿胀。此外，我们表明与YYY-nock-In-In-In小鼠相比，FFF敲入小鼠的耳朵肿胀显着降低。与此结果一致，在FFF-KNOCK-IN小鼠中，CD3^<+> T细胞和嗜酸性粒细胞的浸润严重降低了。此外，使用实时PCR的实验表明，在FFF-KNOCK-IN小鼠的耳朵组织中，TNF-Alpha mRNA的表达也降低。综上所述，这些结果表明，通过FC Epsilon RIβ链调节肥大细胞激活对于在皮肤皮肤皮肤上暴露后的接触超敏反应至关重要。