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IgE-mediated Chronic Allergic Inflammation (IgE-CAI)
免疫球蛋白介导的慢性过敏性炎症小鼠模型(IgE-CAI)   

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Abstract

This system is one of a chronic allergic inflammation model. A single subcutaneous injection of antigens elicits a delayed-type response manifested by ear swelling. The entire response consists of three phases: Immediate, late, and chronic. The early-phase swelling peak within 1 h after allergen challenge is followed by the late-phase swelling 6-10 h late. Importantly, the ear swelling at the late-phase started on day2 and peak on day4. This late-phase ear swelling response is defined as IgE-mediated chronic allergic inflammation (IgE-CAI) and mainly regulated by basophils.

Materials and Reagents

  1. Mice (C57BL/6, BALB/C)
  2. Ascites of the trinitrophenyl (TNP)-specific IgE mAb from the IGELb4 (ATCC, catalog number: TIB141 )
  3. TNP11-OVA (BioTechniques, catalog number: T-5051-10 )
  4. Ovalbumin (OVA) (grade V) (Sigma Aldrich, catalog number: A5503 )
  5. Phosphate buffered saline (PBS) (Life Technologies, Gibco®, catalog number: 20012 )

Equipment

  1. Needle (29G1/2)
  2. Syringe (1 ml) (Terumo Medical Corporation, catalog number: SS-10M2913 )
  3. Dial thickness guage caliper (Ozaki, catalog number: G-1A )

Procedure

  1. Mice are sensitized with TNP-IgE mAb intravenous injection of ascites (200 μl).
  2. One day later, the left ears of mice are injected subcutaneously with 10 μg of TNP11-OVA in 10 μl of PBS.
  3. And their right ears are injected with equal amount of OVA as a control.
  4. Ear thickness is measured with a dial thickness guage caliper for 7 days.

Acknowledgments

This protocol was developed and implemented by Dr. Masato Kubo at Division of Molecular Pathology, Tokyo University of Science, Chiba, Japan and Dr. Hiromasa Inoue at Department of Pulmonary Medicine, Kagoshima University, Kagoshima, Japan. This work was supported by a Grant-in-Aid-of-Scientific Research in Priority Areas of the Ministry of Education, Culture, Sports, Science, and Technology of Japan and the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation.

References

  1. Mukai, K., Matsuoka, K., Taya, C., Suzuki, H., Yokozeki, H., Nishioka, K., Hirokawa, K., Etori, M., Yamashita, M., Kubota, T., Minegishi, Y., Yonekawa, H. and Karasuyama, H. (2005). Basophils play a critical role in the development of IgE-mediated chronic allergic inflammation independently of T cells and mast cells. Immunity 23(2): 191-202.
  2. Sawaguchi, M., Tanaka, S., Nakatani, Y., Harada, Y., Mukai, K., Matsunaga, Y., Ishiwata, K., Oboki, K., Kambayashi, T., Watanabe, N., Karasuyama, H., Nakae, S., Inoue, H. and Kubo, M. (2012). Role of mast cells and basophils in IgE responses and in allergic airway hyperresponsiveness. J Immunol 188(4): 1809-1818.

简介

该系统是慢性过敏性炎症模型之一。 单次皮下注射抗原引起由耳肿胀表现的延迟型反应。 整个反应包括三个阶段:即时,晚期和慢性。 在变应原攻击后1小时内的早期肿胀峰值之后是晚期6-10小时的晚期肿胀。 重要的是,后期的耳肿胀在第2天开始,在第4天达到峰值。 这种晚期耳肿胀反应定义为IgE介导的慢性过敏性炎症(IgE-CAI),并且主要由嗜碱性粒细胞调节。

材料和试剂

  1. 小鼠(C57BL/6,BALB/C)
  2. 来自IGELb4(ATCC,目录号:TIB141)的三硝基苯(TNP) - 特异性IgE mAb的腹水剂
  3. TNP11-OVA(BioTechniques,目录号:T-5051-10)
  4. 卵白蛋白(OVA)(等级V)(Sigma Aldrich,目录号:A5503)
  5. 磷酸盐缓冲盐水(PBS)(Life Technologies,Gibco ,目录号:20012)

设备

  1. 针(29G1/2)
  2. 注射器(1ml)(Terumo Medical Corporation,目录号:SS-10M2913)
  3. 表盘厚度测径器(Ozaki,目录号:G-1A)

程序

  1. 小鼠用TNP-IgE mAb静脉注射腹水(200μl)致敏
  2. 一天后,用10μlPBS中的10μgTNP11-OVA皮下注射小鼠的左耳。
  3. 他们的右耳注射等量的OVA作为对照
  4. 耳厚度用表盘厚度计测量7天。

致谢

该协议由日本千叶县东京科技大学分子病理学部的Masato Kubo博士和日本鹿儿岛鹿儿岛大学肺部医学部Hiromasa Inoue博士开发和实施。 这项工作得到了日本教育,文化,体育,科学和技术部优先领域的科学研究和促进基础研究计划的支持 国家生物医学创新研究所的健康科学。

参考文献

  1. Mukai,K.,Matsuoka,K.,Taya,C.,Suzuki,H.,Yokozeki,H.,Nishioka, K.,Hirokawa,K.,Etori,M.,Yamashita,M.,Kubota,T.,Minegishi, Yonekawa,H。和Karasuyama,H。(2005)。 嗜碱性粒细胞  在IgE介导的慢性发展中起关键作用 过敏性炎症,独立于T细胞和肥大细胞。 免疫 23(2):191-202。
  2. Sawaguchi,M.,Tanaka,S.,Nakatani,Y.,Harada,Y.,Mukai,K.,Matsunaga,Y.,Ishiwata,K.,Oboki,K.,Kambayashi,T.,Watanabe, Karasuyama,H.,Nakae,S.,Inoue,H。和Kubo,M。(2012)。 肥大细胞和嗜碱性粒细胞在IgE反应和过敏性气道高反应性中的作用 J Immunol 188(4):1809-1818。
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Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.
引用:Harada, Y. and Kubo, M. (2012). IgE-mediated Chronic Allergic Inflammation (IgE-CAI). Bio-protocol 2(19): e266. DOI: 10.21769/BioProtoc.266.
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