A noninvasive method for quantifying and distinguishing inflammatory skin reactions☆,☆☆

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Abstract

Background: The ribonuclease protection assay (RPA) represents a technology that allows detection of small amounts of intact RNA. Recent progress in understanding cytokine networks in the skin suggests that measurements of cytokine mRNA levels could provide a method to distinguish various reactions such as irritant contact dermatitis and allergic contact dermatitis that can occur in the skin. Objective: We attempted to differentiate and quantitate irritant and immunologic skin reactions by measuring mRNA levels. Methods: We have used the technique of tape stripping human skin to remove superficial cell layers and have extracted RNA from these skin samples. This RNA was used for RPA analysis. Results: By means of RPA analysis, we have demonstrated distinct cytokine profiles that appear to discriminate, for example, irritant from immunologic skin reactions. Conclusion: We have shown that multiple cytokine mRNA levels can be defined in these RNA samples obtained from the skin. This approach assesses not only the cytokine gene profiles, but at the same time may quantify the severity of common irritant versus allergic skin reactions. (J Am Acad Dermatol 1999;41:687-92.)

Section snippets

Induction of erythematous reactions on the skin

The ICD was induced by applying 0.5% sodium lauryl sulfate (SLS, Fisher Scientific, San Diego, Calif) in distilled water for 72 hours. At this point, the erythema was graded according to a standard scoring scale and tape stripping was performed (see below).12 Dibutyl squarate (2%) in acetone was applied for 48 hours under occlusion to the upper arm to induce the ACD reactions. After about 14 days the dibutyl squarate (2%) was reapplied to elicit the immune-mediated reaction. The scoring of the

Cytokines identified by RPA

We initially used 4 RNA probes (IL-4, IL-8, L32, GAPDH) for hybridization to RNA samples obtained from 1 individual. Fig 1 shows the cytokines identified by RPA analysis of the cells obtained by tape stripping (12 times) skin sites on the upper arms of this individual (the same individual as subject 2 in Fig 2).

. Four RNA probes (IL-4, IL-8, L32, GAPDH) were used for hybridization to RNA samples from 1 individual. Cells obtained by tape stripping (12 times) skin sites on upper arms of this

DISCUSSION

These results demonstrate that skin RNA can be obtained from tape strips without significant degradation as indicated by the ability to detect even low abundant RNA species such as IFN-γ and iNOS (Fig 2). These mRNAs potentially can be characterized in a variety of ways including assessing the presence of cytokine mRNA for various interleukins as was done in this report. The exact species of mRNA probes used in these experiments was determined in part by the necessity for using probes of

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    Reprint requests: Vera B. Morhenn, MD, Clinical Associate Professor, University of California at San Diego, California Skin Research Institute, 15222-B Avenue of Science, San Diego, CA 92128.

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    0190-9622/99/$8.00 + 0  16/1/99779

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    Copyright © 1999 American Academy of Dermatology, Inc. Published by Mosby, Inc. All rights reserved.