Department of Clinical Immunology, Wroclaw Medical University;
Department of Medical Physiology, Chair of Biomedical Sciences, Institute of Physiotherapy, Faculty of Health Sciences, Jagiellonian University Medical College;
J Vis Exp. 2022 Sep 26(187). doi: 10.3791/64329.
Contact hypersensitivity (CHS) is an experimental model of allergic contact dermatitis (ACD) that can be studied in mice. This study aims to present an objective laboratory method that may help to study the CHS reaction in mice, which can be measured and quantified by various tests. To induce CHS, on day "0", mice were sensitized on a previously shaved spot by abdominal skin painting with the hapten 2,4,6-trinitrochlorobenzene (TNCB) in an acetone-ethanol mixture, whereas negative control mice were sham sensitized with vehicle alone-acetone-ethanol mixture. On day "4", the baseline ear thickness was measured with a micrometer prior to the elicitation of CHS (challenge) by painting both ears with diluted TNCB both in the test and control groups. After 24 h, the ear swelling was measured with a micrometer. CHS is an example of a T cell-mediated immune response that causes swelling in inflamed tissue, peaking 24 h after the skin challenge with the same hapten. An increase in ear edema correlated with augmented ear weight, myeloperoxidase (MPO) activity, pro-inflammatory cytokine concentration in the ear extracts, increased thickening of the edematous dermis in the histological examination, and ear vascular permeability. There was also an increase in the concentration of TNP-specific IgG1 antibodies in the sera of the test group when compared with the control mice. Additionally, CHS can be successfully transferred with the CHS-effector cells obtained from donors previously sensitized with TNCB. The CHS-effector cells were administered intravenously into naïve recipient mice, which were subsequently challenged with the same diluted hapten. Ear swelling was measured with a micrometer 24 h later.
接触超敏反应 (CHS) 是一种过敏性接触性皮炎 (ACD) 的实验模型，可在小鼠中进行研究。本研究旨在提出一种客观的实验室方法，可能有助于研究 CHS 反应，该反应可以通过各种测试进行测量和量化。为了诱导 CHS，在第“0”天，通过腹部皮肤涂抹在丙酮-乙醇混合物中的半抗原 2,4,6-三硝基氯苯 (TNCB) 将致敏原致敏于先前剃光的斑点，而阴性对照小鼠则单独用载体-丙酮-乙醇混合物进行假致敏。在第“4”天，在诱发 CHS（挑战）之前，用测微计测量基线耳厚度，两组均用稀释的 TNCB 涂抹双耳。24 小时后，用测微计测量耳肿胀。CHS 是 T 细胞介导的免疫反应的一个例子，它会导致炎症组织肿胀，在皮肤受到相同半抗原挑战后 24 小时达到峰值。耳水肿的增加与耳重量增加、耳组织提取中的髓过氧化物酶 (MPO) 活性、促炎细胞因子浓度、水肿真皮增厚以及耳血管通透性增加相关。与对照组相比，实验组血清中 TNP 特异性 IgG1 抗体的浓度也有所增加。此外，用先前用 TNCB 致敏的供体获得的 CHS 效应细胞可以成功转移 CHS。将 CHS 效应细胞静脉内给予用相同稀释半抗原预先致敏的 naive 受体小鼠，然后 24 小时后用测微计测量耳肿胀。
