Triclosan promotes epicutaneous sensitization to peanut in mice
- 1.5k Downloads
Peanut allergy is increasing in prevalence due to unknown factors. A growing body of clinical evidence suggests sensitization to peanut occurs through the skin, supported by findings in mouse models. There is a need to identify environmental factors that promote epicutaneous sensitization to peanut. Triclosan is an antimicrobial found in household products that has been associated with food sensitization in humans. We tested the impact of triclosan on epicutaneous sensitization to peanut, as well as the milk allergen α-lactalbumin (ALA).
We observed that topical triclosan promoted epicutaneous sensitization to both peanut and ALA, and promoted anaphylaxis to peanut.
Our results demonstrate that the mouse model of epicutaneous sensitization to foods is effective for demonstrating the clinically significant impact of environmental factors such as triclosan on food allergy.
KeywordsFood allergy Sensitization Anaphylaxis Peanut Triclosan Epicutaneous
crude peanut extract
IgE-mediated food allergy is becoming more prevalent in the western world. From 1997 to 2007, food allergy prevalence increased by 18 % among children under the age of 18 years old in the United States . Estimates for the rise in peanut and tree nut allergy have been more dramatic, with a greater than threefold increase . There is growing clinical evidence that sensitization to peanut occurs through the skin . We recently showed in mice that epicutaneous peanut exposure could induce sensitization in the absence of adjuvant or skin damage . We showed that peanut had innate adjuvant activity through activation of IL-33-dependent pathways. This epicutaneous sensitization model helps to explain why peanut is comprised of such potent allergens, but does not incorporate risk factors that increase individual susceptibility to peanut allergy.
Environmental factors that could contribute to the increasing prevalence of food allergy include common environmental chemicals. Triclosan, an antimicrobial agent found in consumer products such as hand sanitizer, soap, and shampoo, has been associated with food sensitization in humans . Triclosan enhances sensitization in a model of experimental asthma . Triclosan has also been shown to induce TSLP expression in the skin, and drive a Th2-skewed immune response . We hypothesized that exposure to environmental factors, such as triclosan, could promote individual susceptibility to peanut allergy in the context of epicutaneous peanut exposure, and set out to test this in the mouse model of epicutaneous sensitization.
Defatted crude peanut extract (CPE) was prepared from deshelled roasted peanuts that were ground to a paste, defatted with acetone, dried overnight, and extracted with PBS with protease inhibitor (Roche Life Science, Indianapolis, IN). α-Lactalbumin (ALA) and triclosan were purchased from Sigma-Aldrich (St. Louis, MO).
Mouse model of sensitization
C3H/HeJ mice were acquired from the National Cancer Institute (Frederick, MD). Female mice (4–5 weeks old) were anesthetized, followed by exposure to 50 μL of (5, 50, or 500 μg) CPE or ALA (100 μg) with vehicle (acetone) or triclosan (1 %) on the ear pinnae. The top dose of CPE and the dose of ALA were previously found to induce sensitization without (CPE) and with (ALA) adjuvant . Mice were exposed weekly for 6 exposures, and mice were challenged 1 week after the last exposure. All procedures were approved by the Animal Care and Use Committee of the Icahn School of Medicine at Mount Sinai (Protocol LA11-00273).
Allergen challenge and anaphylaxis measurement
Mice were challenged by intraperitoneal injection with 100 μg of CPE, and were observed following challenge for 30 min. Mice were orally challenged with 50 mg of ALA, followed by intraperitoneal challenge with 50 μg of ALA in those that did not react. In our experience, mice sensitized to ALA will respond to oral challenge while mice sensitized to peanut require systemic (ip) challenge to induce anaphylaxis. Body temperature was recorded before and 30 min after challenge by rectal thermometer (WPI Instruments, Sarasota, FL).
Allergen-specific IgE, IgG1, and IgG2a were measured in serum obtained prior to challenge. IgE was measured by coating ELISA plates with monoclonal anti-IgE (BD Biosciences, Franklin Lakes, NJ) and detecting with DIG-labeled CPE or ALA. IgG1 and IgG2a were measured by directly coating allergens on the plate and detecting with biotinylated anti-IgG1 or IgG2a (BD Biosciences), followed by Avidin-HRP and TMB reagent (eBiosciences, San Diego, CA). A SpectraMax plate reader with background subtraction was used to measure absorbance (450 nm).
Differences between groups were measured by ANOVA, followed by post hoc testing with non-parametric T test (GraphPad Prism, La Jolla, CA).
Impact of triclosan on dose-dependent epicutaneous sensitization of mice to peanut
Impact of triclosan on epicutaneous sensitization to the milk allergen α-lactalbumin
With the prevalence of food allergies increasing to as much as 10 % in the developed world , understanding the underlying cause(s) driving this increase is imperative. Triclosan is extensively used in household consumer products, and is found in more than 75 % of the US population  and is one of the top ten environmental contaminants of American rivers . Here we show that topical application of triclosan had a modest but significant effect on sensitization associated with experimental food allergy. Triclosan reduced the exposure level required to induce sensitization to allergens with inherent adjuvant activity (peanut), and functioned as an adjuvant when used with weaker allergens (ALA).
Epithelial-derived Th2-inducing cytokines such as TSLP and IL-33 play an important role in driving sensitization to foods [4, 12]. We previously showed that peanut could induce production of IL-33 in the skin , while triclosan was previously shown to induce induce TSLP . Triclosan also suppressed IL-33 expression in skin, indicating that it may counteract the effect of peanut. However, in combination we see that triclosan significantly promotes the development of IgE sensitization to peanut.
This brief report extends our previous detailed description of the epicutaneous sensitization model  by demonstrating the utility of the model in studying the impact of environmental factors on susceptibility to sensitization. Our data also importantly provide evidence of causality in the finding in human cohorts that elevated urinary triclosan is associated with food sensitization . The increasing prevalence of peanut allergy is likely due to multiple factors that individually have modest but biologically significant effects. Environmental factors, either single factors or in combination, can be tested for their potential impact on susceptibility to food allergy using this clinically relevant model of sensitization to peanut.
ST performed all experiments under the supervision of LT. LT and MCB designed experiments. ST and MCB prepared the manuscript, which was read by all authors prior to submission. All authors read and approved the final manuscript.
We thank Jessica Savage for discussions that prompted this work. Supported by NIH R01AI093577, the David H. and Julia Koch Research Program in Food Allergy Therapeutics, and the Robin Chemers Neustein Postdoctoral Fellowship (to LT).
The authors declare that they have no competing interests.
- 3.Brough HA, Simpson A, Makinson K, Hankinson J, Brown S, Douiri A, et al. Peanut allergy: effect of environmental peanut exposure in children with filaggrin loss-of-function mutations. J Allergy Clin Immunol. 2014;134(4):867–75 e861.Google Scholar
- 5.Savage JH, Matsui EC, Wood RA, Keet CA. Urinary levels of triclosan and parabens are associated with aeroallergen and food sensitization. J Allergy Clin Immunol. 2012;130(2):453–60 e457.Google Scholar
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.