Delayed‐Type Hypersensitivity

Yi Luo1, Martin E. Dorf1

1 Harvard Medical School, Boston, Massacusetts
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 4.5
DOI:  10.1002/0471142735.im0405s55
Online Posting Date:  May, 2001
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Abstract

Delayed‐type hypersensitivity (DTH) is an in vivo assay of cell‐mediated immune function. DTH reactions are often divided into two phases: the sensitization phase, referring to the initial immunization with specific antigen, and the efferent or challenge phase of the DTH response, which usually follows 6 to 14 days after sensitization. This unit details the protocol for stimulating DTH responses in mice to the O‐succinimide ester of the hapten 4‐hydroxy‐3‐nitrophenyl acetyl (abbreviated NP‐O‐Su).

     
 
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Table of Contents

  • Reagents and Solutions
  • Commentary
  • Figures
     
 
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Materials

Basic Protocol 1:

  Materials
  • Female B6AF1 hybrid mice (Jackson Labs), 8 to 12 weeks old; four or five per experimental or control group
  • Ketamine/xylazine anesthesian for mice (unit 4.4)
  • NP‐O‐Su solution
  • Borate‐buffered saline, pH 8.6 unit 2.7)
  • Phosphate‐buffered saline, pH 7.8 (PBS; appendix 2A)
  • 0.1 ml Luer‐tip Hamilton syringe (Fisher #14‐824‐11)
  • 27‐G needles
  • 0.01‐ to 10.0‐mm caliper‐type engineer's micrometer (Swiss Precision Instruments) or Peacock dial thickness gauge (Model G‐1A, Ikemoto Scientific Technology; see Figs. 4.2.1A and 4.5.1C)
  • Additional reagents and equipment for anesthesia (unit 1.4) and intraperitoneal and subcutaneous injection of mice (unit 1.6)
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Figures

Videos

Literature Cited

Literature Cited
   Dorf, M.E., Collins, M., Whitters, M.J., O'Hara, R.M., Kelleher, K., Jayaraman, S., and Kuchroo, V.K. 1989. Antigen specific suppressor cells, receptors, and factors. Prog. Immunol. 7:874‐881.
   Mosmann, T.R. and Coffman, R.L. 1989. Th1 and Th2 cells: Different patterns of lymphokine secretion lead to different functional properties. Annu. Rev. Immunol. 7:145‐173.
   Oppenheim, J.J., Zachariae, C.O.C., Mukaida, N., and Matsushima, K. 1991. Properties of the novel proinflammatory supergene “Intercrine” cytokine family. Annu. Rev. Immunol. 9:617‐648.
   Sell, S. 1987. Immunology, Immunopathology and Immunity, 4th ed. Elsevier Science Publishing, New York.
   Sunday, M.E., Weinberger, J.Z., Benacerraf, B., and Dorf, M.E. 1980a. Hapten‐specific T cell responses to 4‐hydroxy‐3‐nitrophenyl acetyl. IV. Specificity of cutaneous sensitivity responses. J. Immunol. 125:1601‐1605.
   Sunday, M.E., Benacerraf, B., and Dorf, M.E. 1980b. Hapten‐specific T cell responses to 4‐hydroxy‐3‐nitrophenyl acetyl. VI. Evidence for different T cell receptors in cells that mediate H‐2I‐restricted and H‐2D‐restricted cutaneous sensitivity responses. J. Exp. Med. 152:1554‐1562.
   Vadas, M.A., Miller, J.F.A.P., 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.
   Zamvil, S.S. and Steinman, L. 1990. The T lymphocyte in experimental allergic encephalomyelitis. Annu. Rev. Immunol. 8:579‐621.
Key Reference
   Sunday, M.E., Weinberger, J.Z., Benacerraf, B., and Dorf, M.E. 1980a. See above.
  Original description of this procedure including a comparison with DTH induced by protein conjugates in CFA.
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