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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Abstract

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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Materials

Basic Protocol 1:  

  Materials
  • 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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Figures

Videos

Literature Cited

Literature Cited
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  Donovan, J. and Brown, P. 2006. Handling and Restraint. Curr. Protoc. Immunol. 73:1.3:1.3.1‐1.3.6.
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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.
  Kaplan, D.H. 2010. In vivo function of Langerhans cells and dermal dendritic cells. Trends Immunol. 31:446‐451. doi: 10.1016/j.it.2010.08.006.
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  Kaplan, D.H., Igyártó, B.Z., and Gaspari, A.A. 2012. Early immune events in the induction of allergic contact dermatitis. Nat. Rev. Immunol. 12:114‐124.
  Luo, Y. and Dorf, M. E. 2001. Delayed‐Type Hypersensitivity. Curr. Protoc. Immunol. 55:4.5.1‐4.5.5.
  Martin SF. 2015. New concepts in cutaneous allergy. Contact Dermatitis 72:2‐10. doi: 10.1111/cod.12311.
  Martin, S.F., Dudda, J.C., Bachtanian, E., Lembo, A., Liller, S., Dürr, C., Heimesaat, M.M., Bereswill, S., Fejer, G., Vassileva, R., Jakob, T., Freudenberg, N., Termeer, C.C., Johner, C., Galanos, C., and Freudenberg, M.A. 2008. Toll‐like receptor and IL‐12 signaling control susceptibility to contact hypersensitivity. J. Exp. Med. 205:2151‐2162. doi: 10.1084/jem.20070509.
  Martin, S.F., Esser, P.R., Schmucker, S., Dietz, L., Naisbitt, D.J., Park, B.K., Vocanson, M., Nicolas, J.F., Keller, M., Pichler, W.J., Peiser, M., Luch, A., Wanner, R., Maggi, E., Cavani, A., Rustemeyer, T., Richter, A., Thierse, H.J., and Sallusto, F. 2010. T‐cell recognition of chemicals, protein allergens and drugs: Towards the development of in vitro assays. Cell. Mol. Life Sci. 67:4171‐4184. doi: 10.1007/s00018-010-0495-3.
  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.
  Phanuphak, P. Moorhead, J.W., and Claman, H.N. 1974. Tolerance and contact hypersensitivity to DNFB in mice. J. Immunol. 112:115‐123.
  Tamaki, T., Fujiwara, H., and Katz, S.I. 1981. The role of epidermal cells in the induction and suppression of contact hypersensitivity. J. Invest. Dermatol. 76:275‐278. doi: 10.1111/1523-1747.ep12526115.
  Toews, G.B., Bergstresser, P.R., and Streilein, J.W. 1980. Epidermal Langerhans cell density determines whether contact hypersensitivity or unresponsiveness follows skin painting with DNFB. J. Immunol. 124:445‐453.
  Vadas, M.A., Miller, J.F., Gamble, J., and Whitelaw, A. 1975. A radioisotopic method to measure delayed type hypersensitivity in the mouse. I. Studies in sensitized and normal mice. Int. Arch. Allergy Appl. Immunol. 49:670‐692. doi: 10.1159/000231449.
  Vocanson, M., Hennino, A., Rozières, A., Poyet, G., and Nicolas, J.F. 2009. Effector and regulatory mechanisms in allergic contact dermatitis. Allergy 64:1699‐1714. doi: 10.1111/j.1398-9995.2009.02082.x.
  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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