A Technique to Assess the Immune Response against an Antigen by Inducing Skin Inflammation

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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Induction of Skin Inflammation in pIL1-DsRed Mice

1. Anesthetize mouse by intraperitoneal (i.p.) injection of an anesthetic cocktail containing 100 mg/kg ketamine, 10 mg/kg xylazine, and 1 mg/kg acepromazine. Adequately anesthetized mice do not show hind paw withdrawal to toe pinch.
2. Apply hair-removal cream to the dorsal surface of both ears of the mouse. Wait 30 sec to 1 min. Wipe ear surface with water-wetted cotton and allow to air-dry.
3. Measure the ear thickness using a micrometer and replace the mouse in a separate cage. Observe until recovery from anesthesia (~ 15 - 30 min). To resolve skin inflammation induced by the hair removal product, allow the mouse to rest for 3 days before further experimentation is carried out.
4. Prepare 1.25% (w/v) oxazolone (OX) by dissolving 16.7 mg of OX in 1 ml acetone and mixing 750 µl of OX/acetone solution with 250 µl of olive oil. Prepare vehicle solution by mixing 750 µl of acetone with 250 µl of olive oil.
5. Re-anesthetize mouse by intraperitoneal (i.p.) injection of anesthetic cocktail (1.1). Measure the ear thickness as before.
6. Apply 12.5 µl of 1.25% OX onto each side of the right ear. Apply 12.5 µl of acetone/olive oil vehicle solution onto each side of the left ear.
7. Keep the mouse separate from cage-mates until fully recovered to prevent insult to its ears by other mice, then replace the mouse in its original cage and return it to the animal housing facility.
   NOTE: Hair removal may result in minor skin inflammation following by change of ear thickness. This minor inflammation will be resolved 3 days later confirmed by normal ear thickness. After topical application for 24 hr, OX-treated ears show significant (p < 0.01) swelling compared to vehicle solution alone-treated ears.

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자료

Name	Company	Catalog Number	Comments
Ketamine hydrochloride	Hospira	NDC 0409-2051-05
Xylazine	LLOYD Laboratory	NADA #139-236
Acepromazine	Boehringer Ingelheim	ANADA 200-361
Hair-removal cream	Church & Dwight
Acetone	Fisher Scientific	A16P4
Oxazolone	Sigma-Aldrich	E0753