Glaspole I N, de Leon M P, Rolland J M, O'Hehir R E
Department of Allergy, Immunology and Respiratory Medicine, Alfred Hospital, Melbourne, Australia.
Allergy. 2005 Jan;60(1):35-40. doi: 10.1111/j.1398-9995.2004.00608.x.
The development of safe and effective immunotherapy for peanut allergy has been complicated by the high anaphylactic potential of native peanut extracts. We sought to map the T-cell epitopes of the major peanut allergen, Ara h 2 in order to develop T-cell targeted vaccines.
A panel of eight peanut-specific CD4+ T-cell lines (TCL) was derived from eight peanut-allergic subjects and proliferative and cytokine responses to stimulation with a set of overlapping 20-mer peptides representing the entire sequence of Ara h 2 determined. Proliferation was assessed in 72 h assays via tritiated thymidine incorporation, while interleukin (IL)-5 and interferon (IFN)-gamma production were assessed via sandwich enzyme-linked immunosorbent assay (ELISA) of cell culture supernatants.
Eight of the 17 Ara h 2 peptides were recognized by one or more subjects, with the two peptides showing highest reactivity [Ara h 2 (19-38) and Ara h 2 (73-92)] being recognized by three subjects each. Adjoining peptides Ara h 2 (28-47) and Ara h 2 (100-119) induced proliferative responses in two subjects. Each of these peptides was associated with a Th2-type cytokine response.
Two highly immunogenic T-cell reactive regions of Ara h 2 have been identified, Ara h 2 (19-47) and Ara h 2 (73-119), providing scope for the development of safe forms of immunotherapy for peanut allergy.
由于天然花生提取物具有较高的过敏反应风险,开发安全有效的花生过敏免疫疗法变得复杂。我们试图绘制主要花生过敏原Ara h 2的T细胞表位,以开发针对T细胞的疫苗。
从8名花生过敏受试者中获得一组8个花生特异性CD4 + T细胞系(TCL),并测定其对一组代表Ara h 2全序列的重叠20肽刺激的增殖和细胞因子反应。通过氚化胸腺嘧啶核苷掺入在72小时试验中评估增殖,而通过细胞培养上清液的夹心酶联免疫吸附测定(ELISA)评估白细胞介素(IL)-5和干扰素(IFN)-γ的产生。
17个Ara h 2肽中的8个被一个或多个受试者识别,其中两个反应性最高的肽[Ara h 2(19 - 38)和Ara h 2(73 - 92)]分别被三个受试者识别。相邻肽Ara h 2(28 - 47)和Ara h 2(100 - 119)在两个受试者中诱导增殖反应。这些肽中的每一个都与Th2型细胞因子反应相关。
已鉴定出Ara h 2的两个高免疫原性T细胞反应区域,即Ara h 2(19 - 47)和Ara h 2(73 - 119),为开发安全形式的花生过敏免疫疗法提供了空间。