Lab Anim Res.  2014 Sep;30(3):95-103. 10.5625/lar.2014.30.3.95.

Characterization of allergic response induced by repeated dermal exposure of IL-4/Luc/CNS-1 transgenic mice to low dose formaldehyde

• Kwak MH
• Kim JE
• Go J
• Koh EK
• Song SH
• Sung JE
• Yang SY
• An BS
• Jung YJ
• Lee JH
• Lim Y
• Hwang DY

• KMID: 2312120
• DOI: http://doi.org/10.5625/lar.2014.30.3.95

Abstract

Although formaldehyde (FA) is known to be a major allergen responsible for allergic contact dermatitis, there are conflicting reports regarding correlation between FA exposure and interleukin (IL-4) expression. To investigate whether allergic responses including IL-4 expression were induced by repeated dermal exposure to low dose FA, alterations in the luciferase signal and allergic phenotypes were measured in IL-4/Luc/CNS-1 transgenic (Tg) mice containing luciferase cDNA under control of the IL-4 promoter after exposure to 4% FA for 2 weeks. High levels of luciferase were detected in the abdominal region of the whole body and submandibular lymph node (SLN) of FA treated mice. Additionally, the ear thickness and IgE concentration were significantly upregulated in the FA treated group when compared with the acetone olive oil (AOO) treated group. FA treated mice showed enhanced auricular lymph node (ALN) weight, epidermis and dermis thickness, and infiltration of inflammatory cells. Furthermore, the expression of IL-6 among T helper 2 cytokines was higher in the FA treated group than the AOO treated group, while vascular endothelial growth factor (VEGF) levels remained constant. Overall, the results presented herein provide additional evidence that various allergic responses may be successfully induced in IL-4/Luc/CNS-1 Tg mice after exposure to low dose FA for 2 weeks. The luciferase signal under the IL-4 promoter may reflect general indicators of the allergic response induced by exposure to low dose FA.

Keyword

Formaldehyde; IL-4; IL-4/Luc/CNS-1 transgenic mice; luciferase signal; IgE

MeSH Terms

Acetone
Animals
Cytokines
Dermatitis, Allergic Contact
Dermis
DNA, Complementary
Ear
Epidermis
Formaldehyde*
Immunoglobulin E
Interleukin-4
Interleukin-6
Interleukins
Luciferases
Lymph Nodes
Mice
Mice, Transgenic*
Olea
Phenotype
Vascular Endothelial Growth Factor A
Olive Oil
Acetone
Cytokines
DNA, Complementary
Formaldehyde
Immunoglobulin E
Interleukin-4
Interleukin-6
Interleukins
Luciferases
Vascular Endothelial Growth Factor A

Figure

• 

  Figure 1 Detection of luciferase signal in the whole body (A) and each organ (B) of IL-4/Luc/CNS-1 Tg mice. Mice were treated with AOO and FA for 2 weeks and then imaged at 24 h after final treatment using the LivingImage software. The color overlay on the image represents the photons per second emitted from the organs in accordance with the pseudocolor scale shown next to the image. In this image, red indicates the highest number of photons per second, while blue indicates the lowest. L, lung; K, kidney; S, spleen; H, heart; SLN, submandibular lymph node; MLN, mesenteric lymph node; T, thymus; P, pancreas. (C) The expression of mouse IL-4 mRNA in SLN was measured by RT-PCR using a specific primer. Data shown are the means±SD (n=5). a, P<0.05 compared to the AOO treated group.

• 

  Figure 2 Alteration of ear phenotypes and IgE concentration. After repeated application of FA solution (4%) to the ear of IL-4/Luc/CNS-1 Tg mice for 2 weeks, the outline of the ear vein in two groups was analyzed using the picture image (A and B), and ear thickness of the mice was measured with a thickness gauge on each experimental day (C). Arrows indicate the vein in the ear. The serum used to measure the IgE concentration was prepared from blood samples collected from the abdominal vein of the mice. The IgE concentration in serum was quantified by an enzyme-linked immunosorbent assay (D). Data shown are the means±SD (n=5). a, P<0.05 compared to the AOO treated group.

• 

  Figure 3 Histological alteration of ear skin. FA solution (4%) was repeatedly applied to the dorsum of the ear of IL-4/Luc/CNS-1 Tg mice. After 2 weeks, ear tissues were collected from AOO or FA treated IL-4/Luc/CNS-1 Tg mice. The histological changes were determined as described in the Materials and Methods. The slide sections of ear tissue were stained with hematoxylin & eosin and then observed at 100× or 400× magnification (Aa-d and Ba-d). Slide sections of ear tissue were stained with toluidine blue and then observed at 400× magnification (Ae and f, and Bc). Arrow heads indicate the infiltrated mast cells in the dermis of the ear. Data shown are the means±SD (n=5). a, P<0.05 compared to the AOO treated group.

• 

  Figure 4 IL-6 and VEGF expression in ear tissue. After detection of specific bands with two primary antibodies, the intensity of each band was determined using an imaging densitometer and the relative level of each protein was calculated based on the intensity of actin protein as an endogenous control. Data represent the mean±SD from three replicates. a, P<0.05 compared to the AOO-treated group.

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