Krasteva M, Peguet-Navarro J, Moulon C, Courtellemont P, Redziniak G, Schmitt D
Laboratoire Peau Humaine et Immunité, INSERM U346, Hôpital E. Herriot, Lyon, France.
Clin Exp Allergy. 1996 May;26(5):563-70.
The need to develop predictive tests which could identify potential allergens has been recognized for many years. There is as yet no accepted in vitro method for the assessment of contact sensitizers.
We have tested the ability of a range of contact allergens to induce in vitro primary sensitization of autologous T cells.
T-cell proliferation induced by haptens using 2-day cultured human Langerhans cells as antigen-presenting cell was assessed by 3H thymidine incorporation. Antigen specific stimulation was calculated as stimulation indexes.
Strong allergens induced in vitro a primary T-cell response in all (trinitrophenyl, TNP: 13/13) or in the majority (fluorescein isothiocyanate, FITC: 7/10) of experiments. An irritant, sodium dodecyl sulfate (SDS), failed to generate a significant T-cell proliferation in any of the experiments (0/10). We obtained a significant lymphoproliferative response to weak sensitizers only in a limited number of experiments: (coumarin: 1/12, citronellal: 0/10, hydroxycitronellal: 2/8). p-Phenylenediamine (PPDA), a prohapten and highly sensitizing chemical in vivo, generated primary sensitization in vitro in only one of six experiments, while Bandrowski's base (BB), a metabolization product of PPDA induced a significant T-cell response in all six experiments.
The present in vitro model allows discrimination between two groups of substances: strong contact sensitizers (TNP, FITC, BB) on the one hand and weak sensitizers (coumarin, citronellal and hydroxycitronellal) and irritants (SDS) on the other hand. It could be used as a screening in vitro assay to eliminate strong contact allergens before further predictive animal tests have to be performed.
多年来人们已经认识到需要开发能够识别潜在过敏原的预测性检测方法。目前尚无被广泛接受的体外评估接触性致敏剂的方法。
我们测试了一系列接触性变应原诱导自体T细胞体外初次致敏的能力。
以培养2天的人朗格汉斯细胞作为抗原呈递细胞,通过³H胸腺嘧啶核苷掺入法评估半抗原诱导的T细胞增殖。抗原特异性刺激以刺激指数计算。
在所有实验(三硝基苯,TNP:13/13)或大多数实验(异硫氰酸荧光素,FITC:7/10)中,强变应原在体外诱导了初次T细胞反应。一种刺激物十二烷基硫酸钠(SDS)在任何实验中均未产生显著的T细胞增殖(0/10)。仅在有限数量的实验中,我们获得了对弱致敏剂的显著淋巴细胞增殖反应:(香豆素:1/12,香茅醛:0/10,羟基香茅醛:2/8)。对苯二胺(PPDA),一种体内前半抗原和高致敏性化学物质,仅在六个实验中的一个实验中在体外产生了初次致敏,而PPDA的代谢产物班德罗夫斯基碱(BB)在所有六个实验中均诱导了显著的T细胞反应。
目前的体外模型能够区分两组物质:一方面是强接触性致敏剂(TNP、FITC、BB),另一方面是弱致敏剂(香豆素、香茅醛和羟基香茅醛)和刺激物(SDS)。它可作为一种体外筛选检测方法,在必须进行进一步的预测性动物试验之前排除强接触性变应原。