Beeton Christine, Chandy K George
Department of Physiology and Biophysics, University of California, Irvine, USA.
J Vis Exp. 2007 Oct 1(8):325. doi: 10.3791/325.
Delayed type hypersensitivity (DTH) is an inflammatory reaction mediated by CCR7- effector memory T lymphocytes that infiltrate the site of injection of an antigen against which the immune system has been primed. The inflammatory reaction is characterized by redness and swelling of the site of antigenic challenge. It is a convenient model to determine the in vivo efficacy of immunosuppressants. Cutaneous DTH can be induced either by adoptive transfer of antigen-specific T lymphocytes or by active immunization with an antigen, and subsequent intradermal challenge with the antigen to induce the inflammatory reaction in a given skin area. DTH responses can be induced to various antigens, for example ovalbumin, tuberculin, tetanus toxoid, or keyhole limpet hemocyanin. Such reactions can also be induced against autoantigen, for example to myelin basic protein (MBP) in rats with experimental autoimmune encephalomyelitis induced with MBP, an animal model for multiple sclerosis (1). Here we demonstrate how to induce an adoptive DTH reaction in Lewis rats. We will first stimulate ovalbumin-specific T cells in vitro and inject these activated cells intraperitoneally to naive rats. After allowing the cells to equilibrate in vivo for 2 days, we will challenge the rats with ovalbumin in the pinna of one ear, while the other ear wil receive saline. The inflammatory reaction will be visible 3-72 hours later and ear thickness will be measured as an indication of DTH severity.
迟发型超敏反应（DTH）是一种由CCR7效应记忆T淋巴细胞介导的炎症反应，这些淋巴细胞会浸润到免疫系统已被致敏的抗原注射部位。炎症反应的特征是抗原攻击部位出现红肿。它是一种用于确定免疫抑制剂体内疗效的便捷模型。皮肤DTH可以通过抗原特异性T淋巴细胞的过继转移或用抗原进行主动免疫，随后用该抗原进行皮内攻击以在给定皮肤区域诱导炎症反应来诱发。DTH反应可针对多种抗原诱发，例如卵清蛋白、结核菌素、破伤风类毒素或钥孔戚血蓝蛋白。此类反应也可针对自身抗原诱发，例如在由髓鞘碱性蛋白（MBP）诱发实验性自身免疫性脑脊髓炎的大鼠中针对MBP诱发，MBP是多发性硬化症的一种动物模型（1）。在此我们展示如何在Lewis大鼠中诱发过继性DTH反应。我们将首先在体外刺激卵清蛋白特异性T细胞，并将这些活化细胞腹腔内注射到未接触过抗原的大鼠体内。让细胞在体内平衡2天后，我们将用卵清蛋白攻击一只耳朵的耳廓，而另一只耳朵注射生理盐水。3至72小时后炎症反应将可见，并且将测量耳朵厚度以作为DTH严重程度的指标。
