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Delayed‐Type Hypersensitivity
Publication Name:
Current Protocols in Immunology
Unit Number:
Unit 4.5
DOI:
10.1002/0471142735.im0405s55
Online Posting Date:
May, 2001 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).
Materials
Basic Protocol
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
- NP-O-Su solution
- Borate-buffered saline, pH 8.6 unit
- 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
- Additional reagents and equipment for anesthesia (unit
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Figures
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Figure 4.5.1(A)Footpad injection for DTH challenge. The needle is inserted above the toes at a 30° angle to deliver 25 µl of solution. (B) Proper placement of thickness gauge for measuring footpad swelling. Note the relative position to the heel. It is imperative that measurements be taken from the same relative position on the foot of the mouse. In this illustration the right “control” footpad is being measured. Swelling can be visualized in the “experimental” contralateral footpad (see arrow). (C) Measuring footpad thickness with a dial thickness gauge. Support and straighten hind limbs by applying pressure beneath the mouse's leg to maintain the limb in a straightened position. The tail should be held down. Consistent holding and positioning of the mouse will facilitate placement of calipers for measurements of footpad thickness.
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 | |
| Original description of this procedure including a comparison with DTH induced by protein conjugates in CFA. | |
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