Name: A Mouse Ear Model for Allergic Contact Dermatitis Evaluation (Video) | JoVE Journal
Uploaded: 2023-03-24T14:55:00.000+00:00
Duration: 8 min 2 s
Description: Learn to induce allergic contact dermatitis in mouse ears using DNFB for immunology and dermatology research. This reproducible protocol enables preclinical evaluation of new therapeutic methods.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

This work was supported by the National Natural Science Foundation of China (NSFC) to N.-N. Y. (81904212);&#160;Jiangsu Traditional Chinese Medicine Science and Technology Project (YB201995); and the Special Funding Project for Postdoctoral Researchers in China (2020T130562).

All the care and treatment of the mice were in accordance with the guidelines established by the Institutional Animal Care and Use Committee of Yangzhou University and were approved by the Institutional Animal Care and Use Committee under the project license SYXK(SU)2022-0044. BALB/c male and female mice aged 6-8 weeks were used in this study. Each group consisted of six mice (see Table of Materials). Cages were placed in a temperature-controlled chamber (22 &#177; 2 &#176;C, 12 h light/dark cycle) with free access to food and water. An experimental flow diagram is shown in Figure 1.
1. Animal preparation
<ol>
	<li>Start the modeling after 1 week of acclimatization to the environment.</li>
	<li>Use an ultraviolet lamp and 75% alcohol disinfectant to clean and disinfect the environment and countertops prior to manipulating the mice. 
	NOTE: In order to avoid the influence of external factors, marking the mice for identification cannot be performed on the mouse ear; staining on the back or on the tail can be used as an alternative.</li>
	<li>Use a small cotton swab to apply soapy water to the abdomen of the mice (approximately 1-2 cm2 in size). Shave the area in the direction of hair growth with a blade or shaver (see Table of Materials) at the beginning of modeling (day 0; Figure 2A). 
	NOTE:&#160;The use of a straight razor blade for hair removal requires a skilled operator. If not performed correctly, it might cause skin irritation. Consider using depilatory cream, clippers, or a safety razor for hair removal.</li>
	<li>Weigh the mouse and compare the weight changes among each group.</li>
</ol>
2. Abdominal sensitization stimulation
<ol>
	<li>Ensure the full recovery of any minor injury to the abdomen skin induced by shaving. Apply abdominal sensitization 2 days after shaving (day 2).</li>
	<li>Prepare the 0.5% DNFB solution: dilute DNFB with an acetone:olive oil mixture at a 4:1 ratio (e.g., 400 &#181;L of acetone mixed with 100 &#181;L of olive oil; see Table of Materials). Use a pipette gun to blow and mix 20 times to thoroughly mix the DNFB solution. Before each administration of DNFB solution to the mouse, blow and mix it three to five times. 
	NOTE: Prepare the solution before use and wrap it in aluminum foil to protect it from direct sunlight.</li>
	<li>Apply 25 &#181;L of the 0.5% DNFB solution to the skin of the shaved area on the abdomen of the mice with a pipettor (Figure 2B).</li>
	<li>Dribble the DNFB solution over the middle of the abdominal shaving area and lightly spread with the smooth side of the pipettor tip to uniformly disperse it.</li>
	<li>At 30 s after DNFB stimulation, place the mice in empty cages without bedding to prevent them from rubbing off the DNFB solution. When the DNFB solution is completely dry (about 2 min), return the mice to their original cage.</li>
	<li>Wear gloves when handling the DNFB solution as it is strongly irritating to human skin.</li>
</ol>
3. Ear sensitization stimulation
<ol>
	<li>Prepare a 0.2% DNFB solution as above, the vehicle solution (a 4:1 mixture of acetone and olive oil), and pure water.</li>
	<li>Orient the mouse body and make the outside border of the auricle face downward during the entire operation to prevent the solution from entering the ear canal during DNFB stimulation.</li>
	<li>On days 4, 6, 8, and 10, use a pipettor to apply 20 &#181;L of the 0.2% DNFB solution or vehicle solution slowly and uniformly to the inner surface of the left auricles of the mice. To avoid DNFB solution from entering the ear canal, use the smooth side of the pipettor tip to gently distribute the DNFB solution during administration. Leave the right ears untreated (Figure 2C).</li>
	<li>Wait until the DNFB solution is dry and place the mice back in the cage (about 30 s).</li>
	<li>Wear gloves when handling the DNFB solution.</li>
</ol>
4. Recording mouse weight and ACD symptoms
<ol>
	<li>Weigh the mouse every day, start&#160;on day 1, and compare with its corresponding weight on day 0;&#160;evaluate the effect of ACD on the body weight of mice as weight change (g) &#177; standard error of the mean (SEM).</li>
	<li>Take high resolution photos of the mouse ears to record ACD clinical symptoms every 2 days, starting on day 1.</li>
</ol>
5. Measurement of the auricle thickness
<ol>
	<li>Measure the auricle thickness every 2 days, starting on day 1. Measure and record both ears in detail.</li>
	<li>Use vernier calipers (see Table of Materials) to measure the auricle thickness at the same time each day for accurate results (Figure 3A). Stop the vernier calipers from continuing inward clamping when there is a slight blockage, to prevent tissue damage to the mouse ear. Keep the position fixed and record the data.</li>
	<li>Collect the thickness from three different locations on each auricle (Figure 3B). Record the average of the three data as a valid value. Evaluate the ear swelling in micrometers (&#181;m) &#177; standard error of the mean (SEM).</li>
</ol>
6. Evaluation of the degree of inflammatory swelling
<ol>
	<li>Prepare 0.5% Evans blue dye (see Table of Materials) solution: dilute Evans blue dye with phosphate buffered saline (PBS) on day 11. Wear a lab coat and gloves at all times, as Evans blue dye is slightly toxic to humans.</li>
	<li>Immobilize the mice with a fixator:&#160;open the lid of the fixator (see Table of Materials),&#160;hold the mouse tail, make the head of the mouse face the fixator, and make the mouse instinctively climb into the fixator. Cover the lid, make the mouse tail come out of the hole on the lid, and adjust the length of the fixator to expose the whole mouse tail.</li>
	<li>Wipe the tail repeatedly with an alcohol cotton ball or soak it in warm water for 30 s, and gently pinch the root of the tail to fill and expand the veins on both sides. Perform the injection under the irradiation of a cold light source.</li>
	<li>Slowly inject Evans blue dye solution into the mouse tail vein using a 1 mm insulin needle. Wait for 15 min and then take pictures of the mouse ears. 
	NOTE: &#8203;Put the mouse on the table and gently hold it&#160;to expose the ear region for image acquisition. Shortly after injection with the Evans blue dye solution and observing&#160;corresponding indications, use cervical dislocation to euthanize the mouse.</li>
</ol>

<ol><li>Neale, H., Garza-Mayers, A. C., Tam, I., Yu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pediatric+allergic+contact+dermatitis.+Part+I%3A+Clinical+features+and+common+contact+allergens+in+children%5BTitle%5D)+AND+%22Journal+of+the+American+Academy+of+Dermatology%22%5BJournal%5D)">Pediatric allergic contact dermatitis. Part I: Clinical features and common contact allergens in children.</a> Journal of the American Academy of Dermatology. 84 (2), 235-244 (2021).</li>
<li>Koppes, S. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Current+knowledge+on+biomarkers+for+contact+sensitization+and+allergic+contact+dermatitis%5BTitle%5D)+AND+%22Contact+Dermatitis%22%5BJournal%5D)">Current knowledge on biomarkers for contact sensitization and allergic contact dermatitis.</a> Contact Dermatitis. 77 (1), 1-16 (2017).</li>
<li>Martin, S. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mechanisms+of+chemical-induced+innate+immunity+in+allergic+contact+dermatitis%5BTitle%5D)+AND+%22Allergy%22%5BJournal%5D)">Mechanisms of chemical-induced innate immunity in allergic contact dermatitis.</a> Allergy. 66 (9), 1152-1163 (2011).</li>
<li>Kimber, I., Basketter, D. A., Gerberick, G. F., Dearman, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Allergic+contact+dermatitis%5BTitle%5D)+AND+%22International+Immunopharmacology%22%5BJournal%5D)">Allergic contact dermatitis.</a> International Immunopharmacology. 2 (2-3), 201-211 (2002).</li>
<li>Vocanson, M., Hennino, A., Rozieres, A., Poyet, G., Nicolas, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effector+and+regulatory+mechanisms+in+allergic+contact+dermatitis%5BTitle%5D)+AND+%22Allergy%22%5BJournal%5D)">Effector and regulatory mechanisms in allergic contact dermatitis.</a> Allergy. 64 (12), 1699-1714 (2009).</li>
<li>Gamradt, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Inhibitory+checkpoint+receptors+control+CD8%2B+resident+memory+T+cells+to+prevent+skin+allergy%5BTitle%5D)+AND+%22The+Journal+of+Allergy+and+Clinical+Immunology%22%5BJournal%5D)">Inhibitory checkpoint receptors control CD8+ resident memory T cells to prevent skin allergy.</a> The Journal of Allergy and Clinical Immunology. 143 (6), 2147.e9-2157.e9 (2019).</li>
<li>Fraginals, R., Blasi, N. A., Lepoittevin, J. P., Benezra, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+successful+murine+model+for+contact+sensitization+to+a+sesquiterpene-alpha-methylene-gamma-butyrolactone%3A+sensitization+to+alantolactone+in+four+strains+of+mice%5BTitle%5D)+AND+%22The+Journal+of+Investigative+Dermatology%22%5BJournal%5D)">A successful murine model for contact sensitization to a sesquiterpene-alpha-methylene-gamma-butyrolactone: sensitization to alantolactone in four strains of mice.</a> The Journal of Investigative Dermatology. 97 (3), 473-477 (1991).</li>
<li>Knop, J., Riechmann, R., Neumann, C., Macher, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Modulation+of+suppressor+mechanisms+in+allergic+contact+dermatitis%3A+5.+Evidence+that+inhibition+of+suppressor+T+lymphocytes+by+Corynebacterium+parvum+is+mediated+by+interferon%5BTitle%5D)+AND+%22The+Journal+of+Investigative+Dermatology%22%5BJournal%5D)">Modulation of suppressor mechanisms in allergic contact dermatitis: 5. Evidence that inhibition of suppressor T lymphocytes by Corynebacterium parvum is mediated by interferon.</a> The Journal of Investigative Dermatology. 79 (6), 385-388 (1982).</li>
<li>Polak, L., Scheper, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Antigen-specific+T-cell+lines+in+DNCB-contact+sensitivity+in+guinea+pigs%5BTitle%5D)+AND+%22The+Journal+of+Investigative+Dermatology%22%5BJournal%5D)">Antigen-specific T-cell lines in DNCB-contact sensitivity in guinea pigs.</a> The Journal of Investigative Dermatology. 80 (5), 398-402 (1983).</li>
<li>Witten, V. H., March, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Studies+of+the+mechanism+of+allergic+eczematous+contact+dermatitis.+II.+Use+of+C14+labelled+2%3A4-dinitrochlorobenzene+in+guinea+pigs%5BTitle%5D)+AND+%22The+Journal+of+Investigative+Dermatology%22%5BJournal%5D)">Studies of the mechanism of allergic eczematous contact dermatitis. II. Use of C14 labelled 2:4-dinitrochlorobenzene in guinea pigs.</a> The Journal of Investigative Dermatology. 31 (2), 97-102 (1958).</li>
<li>Knop, J., Riechmann, R., Macher, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Modulation+of+suppressor+mechanism+in+allergic+contact+dermatitis.+IV.+Selective+inhibition+of+suppressor+T-lymphocytes+by+serum+obtained+from+Corynebacterium+parvum+treated+mice%5BTitle%5D)+AND+%22The+Journal+of+Investigative+Dermatology%22%5BJournal%5D)">Modulation of suppressor mechanism in allergic contact dermatitis. IV. Selective inhibition of suppressor T-lymphocytes by serum obtained from Corynebacterium parvum treated mice.</a> The Journal of Investigative Dermatology. 77 (6), 469-473 (1981).</li>
<li>Rubic-Schneider, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((GPR91+deficiency+exacerbates+allergic+contact+dermatitis+while+reducing+arthritic+disease+in+mice%5BTitle%5D)+AND+%22Allergy%22%5BJournal%5D)">GPR91 deficiency exacerbates allergic contact dermatitis while reducing arthritic disease in mice.</a> Allergy. 72 (3), 444-452 (2017).</li>
<li>Stampf, J. L., Benezra, C., Byers, V., Castagnoli Jr, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Induction+of+tolerance+to+poison+ivy+urushiol+in+the+guinea+pig+by+epicutaneous+application+of+the+structural+analog+5-methyl-3-n-pentadecylcatechol%5BTitle%5D)+AND+%22The+Journal+of+Investigative+Dermatology%22%5BJournal%5D)">Induction of tolerance to poison ivy urushiol in the guinea pig by epicutaneous application of the structural analog 5-methyl-3-n-pentadecylcatechol.</a> The Journal of Investigative Dermatology. 86 (5), 535-538 (1986).</li>
<li>Dhingra, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Molecular+profiling+of+contact+dermatitis+skin+identifies+allergen-dependent+differences+in+immune+response%5BTitle%5D)+AND+%22The+Journal+of+Allergy+and+Clinical+Immunology%22%5BJournal%5D)">Molecular profiling of contact dermatitis skin identifies allergen-dependent differences in immune response.</a> The Journal of Allergy and Clinical Immunology. 134 (2), 362-372 (2014).</li>
<li>Maguire Jr, H. C., Kaidbey, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Experimental+photoallergic+contact+dermatitis%3A+a+mouse+model%5BTitle%5D)+AND+%22The+Journal+of+Investigative+Dermatology%22%5BJournal%5D)">Experimental photoallergic contact dermatitis: a mouse model.</a> The Journal of Investigative Dermatology. 79 (3), 147-152 (1982).</li>
<li>Qiu, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+dysregulated+sebum-microbial+metabolite-IL-33+axis+initiates+skin+inflammation+in+atopic+dermatitis%5BTitle%5D)+AND+%22The+Journal+of+Experimental+Medicine%22%5BJournal%5D)">A dysregulated sebum-microbial metabolite-IL-33 axis initiates skin inflammation in atopic dermatitis.</a> The Journal of Experimental Medicine. 219 (10), e2021397(2022).</li>
<li>Kim, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Anti-inflammatory+activities+of+Dictamnus+dasycarpus+Turcz.%2C+root+bark+on+allergic+contact+dermatitis+induced+by+dinitrofluorobenzene+in+mice%5BTitle%5D)+AND+%22Journal+of+Ethnopharmacology%22%5BJournal%5D)">Anti-inflammatory activities of Dictamnus dasycarpus Turcz., root bark on allergic contact dermatitis induced by dinitrofluorobenzene in mice.</a> Journal of Ethnopharmacology. 149 (2), 471-477 (2013).</li>
<li>Zhou, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effect+of+6%27-acetylpaeoniflorin+on+dinitrochlorobenzene-induced+allergic+contact+dermatitis+in+BALB%2Fc+mice%5BTitle%5D)+AND+%22Immunologic+Research%22%5BJournal%5D)">Effect of 6'-acetylpaeoniflorin on dinitrochlorobenzene-induced allergic contact dermatitis in BALB/c mice.</a> Immunologic Research. 64 (4), 857-868 (2016).</li>
<li>Donglang, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Comparative+study+on+different+skin+pruritus+mouse+models%5BTitle%5D)+AND+%22Frontiers+in+Medicine%22%5BJournal%5D)">Comparative study on different skin pruritus mouse models.</a> Frontiers in Medicine. 8, 630237(2021).</li>
<li>Yang, N., Shao, H., Deng, J., Liu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Network+pharmacology-based+analysis+to+explore+the+therapeutic+mechanism+of+Cortex+Dictamni+on+atopic+dermatitis%5BTitle%5D)+AND+%22Journal+of+Ethnopharmacology%22%5BJournal%5D)">Network pharmacology-based analysis to explore the therapeutic mechanism of Cortex Dictamni on atopic dermatitis.</a> Journal of Ethnopharmacology. 304, 116023(2023).</li>
</ol>

The authors report no conflicts of interest in this work.

The protocol described here for inducing ACD-like symptoms in the ears of mice can be used to study the pathophysiology of ACD and as a screening tool for the development of new drugs.
There are two key steps to establish an ACD model: initial sensitization, and subsequent stimulation. The abdomen is usually the site of initial sensitization, but the subsequent stimulation site was chosen slightly differently. Previous studies have shown that most scholars choose to use chemical sensitizers su...

Allergic contact dermatitis (ACD) is a common skin disease that is characterized by eczema-like symptoms at the contact site, edema and erythema in moderate cases, and papules, erosion, exudation, or even massive scars in severe cases1. It affects up to 20% of the population and can affect people of any age2. ACD often occurs in individuals who have been exposed to allergens repeatedly and can be caused by the individual&#39;s immune response to one or more allergens in their home or workplace3. Type IV delayed hypersensitivity is considered the main type of immune response in ACD4. In areas of the skin that have been repeatedly exposed to allergens, circulating memory T cells accumulate in large numbers and induce immune and inflammatory responses3,5,6. The purpose of this work is to propose a reliable laboratory technique for further investigation of immunological and inflammatory responses in the development of ACD.
The onset of ACD is usually due to contact hypersensitivity caused by repeated exposure to chemicals. Numerous researchers have developed various ACD animal models in house mice7,8, guinea pigs9,10, and other animals over the course of the last few decades, in order to simulate the onset of the disease. Most of the experimental methods consist of two stages: abdominal sensitization (induction) and providing stimuli on the back or the ear lobe (stimulation). Commonly used chemical substances mainly include 1-fluoro-2,4-dinitrobenzene (DNFB)/1-chloro-2,4-dinitrobenzene (DNCB)8,9,11, oxazolone12, urushiol13, etc. Among them, DNFB and DNCB are the most widely used, first reported in October 195810. The nickel sensitization model14 and the photoallergic contact dermatitis model15 are also frequently used.
We present an experimental method for building the ACD model. This method is summarized and optimized on the basis of previous studies and upon comparison with multiple experiments. Compared with other ACD models, this model has some advantages, such as small individual differences, short experimental periods, a small amount of chemical stimulation, etc. In addition, this study is applicable to mice, which are not only economical but also have more options for gene knockout or transgenic mice preparation16. We also describe the various methods used to monitor ACD progress in the experiment, such as measuring ear thickness, using Evans blue dye to measure inflammatory exudation, etc. This model can not only analyze mouse ears, blood, spleen, and other samples by laboratory means to explore the pathogenesis of ACD, but also is applicable for the preclinical evaluation of new therapeutic methods, which has a certain promotional significance.

<table><tbody><tr><td>1-Fluoro-2,4-dinitrobenzene (DNFB)</td><td>Merck</td><td>200-734-3</td><td>1-Fluoro-2,4-dinitrobenzene, &amp;ge;99%</td></tr><tr><td>Acetone</td><td>Sinopharm Chemical Reagent Co. LTD</td><td>10000418</td><td>&amp;ge;99.5%</td></tr><tr><td>Aluminum foil&amp;nbsp;</td><td>Cleanwrap</td><td>CF-2</td><td></td></tr><tr><td>Evans blue dye</td><td>Solarbio</td><td>314-13-6</td><td>Dye content approx. 80%</td></tr><tr><td>Mouse fixator</td><td>ZHUYANBANG</td><td>GEGD-SM1830</td><td></td></tr><tr><td>Olive oil</td><td>Solarbio</td><td>8001-25-0</td><td>500 ml</td></tr><tr><td>Pipet tip</td><td>Biofount</td><td>FT-200</td><td>10 - 200 &amp;mu;l</td></tr><tr><td>Pipettor</td><td>Eppendorf AG</td><td>3123000250</td><td>20 - 200 &amp;mu;l</td></tr><tr><td>Razor blade</td><td>Shanghai Gillette Co. LTD</td><td>74-S</td><td></td></tr><tr><td>Vernier calipers</td><td>Delixi Electric</td><td>DECHOTVCS1200</td><td></td></tr></tbody></table>

a mouse ear model for allergic contact dermatitis evaluation

A Mouse Ear Model for Allergic Contact Dermatitis Evaluation

Skin is the human body&#39;s first line of defense and one of the most exposed organs to environmental chemicals. Allergic contact dermatitis (ACD) is a common skin disease that manifests as a local rash, redness, and skin lesions. The occurrence and development of ACD are influenced by both genetic and environmental factors. Although many scholars have constructed a series of models of ACD in recent years, the experimental protocols of these models are all different, which makes it difficult for readers to establish them well. Therefore, a stable and efficient animal model is of great significance to further study the pathogenesis of atopic dermatitis. In this study, we detail a modeling method using 1-fluoro-2,4-dinitrobenzene (DNFB) to induce ACD-like symptoms in the ears of mice and describe several methods for assessing the severity of dermatitis during modeling. This experimental protocol has been successfully applied in some experiments and has a certain promotional role in the field of ACD research.

Under repeated DNFB stimulation, the mouse ears of the DNFB group displayed evident clinical symptoms comparable to ACD, with sensitive areas showing the typical symptoms of redness, dryness, and even erosion and exudation. However, ear administration of pure water (control group) or solvent control (vehicle group) did not produce similar symptoms (Figure 4).
Meanwhile, in the DNFB group, compared to the untreated right ear, the thickness of the left ear increased...

Here, we describe the methods for inducing allergic contact dermatitis in mouse ears by 1-fluoro-2,4-dinitrobenzene (DNFB) and how to evaluate the severity of allergic contact dermatitis.

Skin is the human body&#39;s first line of defense and one of the most exposed organs to environmental chemicals. Allergic contact dermatitis (ACD) is a ...

a-mouse-ear-model-for-allergic-contact-dermatitis-evaluation

Research

JoVE Journal

Immunology and Infection

Experimental mouse models of allergic asthma offer new possibilities for studying disease pathogenesis and developing new therapeutics. These models are well suited to measuring factors governing the allergic immune response, airway inflammation, and pulmonary pathophysiology.

Murine Model of Allergen Induced Asthma

This video describes a method to measure UV sensitivity in a humanized mouse model—a mouse model that mimics human skin. We determine the minimal erythematous dose (MED) of UV radiation that can cause erythema or edema on the skin. The method can help to find out the efficacy of drugs used to reduce UV sensitivity.

Minimum Erythematous Dose Assay: A Method for Measuring UV Sensitivity in Mouse Models

This video demonstrates an in vivo technique to assess a test antigen by inducing skin inflammation in a murine model. The test antigen—a hapten in an oil/acetone solution—is topically applied to the skin of the mouse ear, and inflammation of the skin is assessed after an adequate period.

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

This video demonstrates Malassezia-induced skin infection in mice. Malassezia oil suspension is applied over the wounded ear, triggering inflammation and swelling of the ear tissue. Later, the ears are dissected, homogenized, and spread on an agar plate for growth. The number of colonies indicates the fungal burden resulting from skin inflammation.

Malassezia-Induced Skin Inflammation and Fungal Burden Analysis in a Murine Model

Allergic responses, characterized by the activation of mast cells and basophils, are driven by the cross-linking of IgE and release of proinflammatory mediators. A quantitative assessment of allergic responses can be achieved by using Evans Blue dye to monitor changes in vascular permeability after allergen challenge.

Measuring Local Anaphylaxis in Mice

Here, we describe a high resolution whole-mount imaging method in the entire adult mouse ear skin, which enables us to visualize branching morphogenesis and patterning of peripheral nerves and blood vessels, as well as immune cell distribution.

Whole-mount Confocal Microscopy for Adult Ear Skin: A Model System to Study Neuro-vascular Branching Morphogenesis and Immune Cell Distribution

An approach is described for real-time detection of the innate immune response to cutaneous wounding and Staphylococcus aureus infection of mice. By comparing LysM-EGFP mice (which possess fluorescent neutrophils) with a LysM-EGFP crossbred immunodeficient mouse strain, we advance our understanding of infection and the development of approaches to combat infection.

A Mouse Model to Assess Innate Immune Response to Staphylococcus aureus Infection

Contact hypersensitivity (CHS) is a murine experimental model of allergic contact dermatitis (ACD). CHS is based on sensitization with reactive hapten by painting the shaved skin of the chest and abdomen, with a subsequent ear skin challenge with a diluted hapten, causing a swelling reaction that is assessed in various ways.

Contact Hypersensitivity as a Murine Model of Allergic Contact Dermatitis

This protocol outlines the procedure for inducing acne inflammation in rat skin with oleic acid and Cutibacterium acnes.

A Rat Model of Compound Acne

The study demonstrates the role of the skin in modulating immune responses in mice through epicutaneous (EC) immunization. EC immunization with a protein antigen can suppress immune responses in contact hypersensitivity or reverse skin-induced suppression when combined with pathogen-associated molecular patterns. EC immunization shows promise as an effective, needle-free approach to immunotherapy.

Murine Model of Epicutaneously-Induced Immunomodulation

Learn to induce allergic contact dermatitis in mouse ears using DNFB for immunology and dermatology research. This reproducible protocol enables preclinical evaluation of new therapeutic methods.

A Mouse Ear Model for Allergic Contact Dermatitis Evaluation

Explore More Videos