Schulten Véronique, Sette Alessandro
Division of Vaccine Discovery, La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA.
Methods Mol Biol. 2018;1799:153-163. doi: 10.1007/978-1-4939-7896-0_13.
Type I allergy is a disease primarily mediated by immunoglobulin E (IgE) and T helper type 2 (Th2) cells. The role of Th2 and other T cell subsets in the pathology of allergic disease as well as induction of tolerance has become an area of intense research over the last decades. Studying allergen-specific T cells to gain a better understanding of their contribution to allergic pathology and how they are modulated by allergen-specific immunotherapy requires knowledge of the allergens targeted by these cells. Identification of T cell epitopes in allergy can be achieved by a variety of methods. In this chapter, we will focus on a technique named FluoroSpot, which relies on the detection of cytokines secreted by T cells in response to stimulation with an antigen (allergen), such as timothy grass (TG) extract or an allergen-derived peptide, for which the cell is specific. We will describe how to overcome the challenge of detecting rare, TG-specific, T cells that occur at low frequency in the blood by using an in vitro expansion culture and subsequent mapping of the precise T cell epitope using FluoroSpot.
I型过敏是一种主要由免疫球蛋白E（IgE）和2型辅助性T细胞（Th2）介导的疾病。在过去几十年中，Th2和其他T细胞亚群在过敏性疾病病理学以及诱导耐受中的作用已成为一个深入研究的领域。研究过敏原特异性T细胞以更好地了解它们对过敏性病理学的贡献以及它们如何被过敏原特异性免疫疗法调节，需要了解这些细胞所靶向的过敏原。过敏中T细胞表位的鉴定可以通过多种方法实现。在本章中，我们将重点介绍一种名为FluoroSpot的技术，该技术依赖于检测T细胞在受到抗原（过敏原）刺激后分泌的细胞因子，例如梯牧草（TG）提取物或细胞特异性的过敏原衍生肽。我们将描述如何通过使用体外扩增培养以及随后使用FluoroSpot对精确的T细胞表位进行定位，来克服检测血液中低频出现的罕见TG特异性T细胞的挑战。
