Christian Doppler Laboratory for Allergy Diagnosis and Therapy, Department of Molecular Biology, University of Salzburg, Salzburg, Austria.
Int Arch Allergy Immunol. 2012;158(4):326-34. doi: 10.1159/000333551. Epub 2012 Apr 3.
Knowledge of allergen-specific T cell epitopes is a prerequisite not only for therapeutic approaches but also for elucidating immunological mechanisms of type I allergy. Ex vivo detection of allergen-specific T cells using class II tetramer technology has become an important tool for investigating immune responses in atopic and healthy individuals.
Using (3)H-thymidine incorporation assays, T cell epitopes specific for the major timothy grass pollen allergen Phl p 5.0101 were mapped in 11 allergic donors and two different mouse strains. Different protocols for expansion/restimulation of T cells from the blood of allergic donors and detection of allergen-specific T cells by Class II Ultimer staining were evaluated.
We identified several new Phl p 5.0101 class II T cell epitopes in allergic patients and confirmed previously published ones. Additionally, we discovered the major T cell epitopes in BALB/c and C57BL/6 mice. Using a novel Class II Ultimer, we detected epitope-specific T cells expanded from the blood of an allergic donor.
Epitope mapping of Phl p 5.0101 revealed an immunodominant epitope in BALB/c and C57BL/6 mice and an immunodominant region in humans (amino acids 259-282), which was recognized by 8 out of 11 allergic donors. Detection of Phl p 5-specific T cells was demonstrated using a Class II Ultimer specific for epitope 196-210. Successful detection of ultimer-positive T cells was strongly dependent on a resting phase after in vitro expansion.
不仅是治疗方法，了解过敏原特异性 T 细胞表位也是阐明 I 型过敏免疫机制的前提。使用 II 类四聚体技术检测过敏原特异性 T 细胞已成为研究变应性和健康个体免疫反应的重要工具。
使用（3）H-胸腺嘧啶掺入测定法，在 11 名过敏供体和两种不同的小鼠品系中绘制主要豚草花粉过敏原 Phl p 5.0101 的特异性 T 细胞表位。评估了用于从过敏供体血液中扩增/再刺激 T 细胞和通过 II 类 Ultimer 染色检测过敏原特异性 T 细胞的不同方案。
我们在过敏患者中鉴定了几个新的 Phl p 5.0101 类 II T 细胞表位，并证实了先前发表的表位。此外，我们还发现了 BALB/c 和 C57BL/6 小鼠中的主要 T 细胞表位。使用新型 II 类 Ultimer，我们从过敏供体的血液中检测到了表位特异性 T 细胞的扩增。
Phl p 5.0101 的表位作图揭示了 BALB/c 和 C57BL/6 小鼠中的免疫优势表位和人类中的免疫优势区域（氨基酸 259-282），这在 11 名过敏供体中的 8 名中得到了识别。使用针对表位 196-210 的 II 类 Ultimer 检测到 Phl p 5 特异性 T 细胞。成功检测 Ultimer 阳性 T 细胞强烈依赖于体外扩增后的休息阶段。
