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IgE-mediated Chronic Allergic Inflammation (IgE-CAI)   

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Original research article

A brief version of this protocol appeared in:
The Journal of Immunology
Feb 2012

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.
Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: 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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