Contact Hypersensitivity

Anthony A. Gaspari1, Stephen I. Katz1, Stefan F. Martin2

1 National Cancer Institute, Bethesda, Maryland, 2 Allergy Research Group, Department of Dermatology, Medical Center, University of Freiburg, Freiburg
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 4.2
DOI:  10.1002/0471142735.im0402s113
Online Posting Date:  April, 2016
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Contact hypersensitivity (CHS) is a simple in vivo assay of cell‐mediated immune function in which exposure of epidermal and dermal cells to exogenous haptens results in a delayed‐type hypersensitivity (DTH) reaction that can be measured and quantified. Epidermal Langerhans cells and dermal dendritic cells are the critical antigen‐presenting cells in this reaction which initiate sensitization to haptens by presenting antigens to CD4‐ and CD8‐bearing T lymphocytes which, in turn, secrete cytokines and recruit other cells to the site of the reaction. In the protocol described here, mice are shaved and the skin of their abdomens is exposed to a hapten. After 5 or 6 days (the afferent phase), the baseline ear thickness is measured prior to initiation of the efferent phase. Finally, the ear is treated epicutaneously with the hapten solution and ear thickness is measured in ∼24 hr. The magnitude of the ear swelling reaction after allergen treatment reflects the strength of the immune response. © 2016 by John Wiley & Sons, Inc.

Keywords: contact hypersensitivity; delayed‐type hypersensitivity (DTH); allergic contact dermatitis

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Table of Contents

  • Basic Protocol 1:  
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
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Basic Protocol 1:  

  • Female mice, 6‐ to 12‐week‐old and pathogen‐free
  • 3% (w/v) TNCB in acetone (see recipe)
  • 1% (w/v) TNCB in acetone (see recipe)
  • Small animal clipper (Oster A‐2)
  • Micropipettor with disposable tips
  • Dial thickness gauge, 0.01 to 12.5 mm (see Fig.  A; Swiss Precision Instruments or Käfer Messuhrenfabrik GmbH & Co. KG)
  • Indelible marking pen
  • Additional equipment for animal handling and restraint (Donovan and Brown, )
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Literature Cited

Literature Cited
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  Furue, M. and Tamaki, K. 1985. Induction and suppression of contact hypersensitivity to FITC. J. Invest. Dermatol. 85:139‐142. doi: 10.1111/1523-1747.ep12276557.
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  Miller, J.F., Vadas, M.A., Whitelaw, A., and Gamble, J. 1975. A radioisotopic method to measure delayed type hypersensitivity in the mouse. II. Cell transfer studies. Int. Arch. Allergy Appl. Immunol. 49:692‐708. doi: 10.1159/000231450.
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  Weber, F.C., Esser, P.R., Müller, T., Ganesan, J., Pellegatti, P., Simon, M.M., Zeiser, R., Idzko, M., Jakob, T., and Martin, S.F. 2010. Lack of the purinergic receptor P2X(7) results in resistance to contact hypersensitivity. J. Exp. Med. 207:2609‐2619. doi: 10.1084/jem.20092489.
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