Guironnet G, Dalbiez-Gauthier C, Rousset F, Schmitt D, Péguet-Navarro J
INSERM U346, Pavillon R, Hôpital E. Herriot, 69374, 03, Lyon, France.
Toxicol In Vitro. 2000 Dec;14(6):517-22. doi: 10.1016/s0887-2333(00)00043-6.
We previously reported that in vitro primary sensitization of hapten-specific T cells by cultured human epidermal Langerhans cells (LC) provides an alternative approach to discriminate strong contact sensitizers from irritants (Krasteva et al., 1996; Moulon et al., 1993). However, this LC-based immunoassay was limited by the availability of human skin samples. In the present study, we used monocyte-derived dendritic cells (DC) to analyse the autologous proliferative T cell response to several allergens. Monocytes were purified from the peripheral blood of healthy donors and cultured for 6-8 days in the presence of GM/CSF and IL-4 and then for 2 days in the presence of GM/CSF and TNFalpha. The resulting cells exhibited the phenotype of mature DC, as assessed by the strong expression of HLA-DR, CD80, CD83 and CD86 antigens. We showed that trinitrophenyl (TNP)-treated mature DC induced a significant T cell proliferative response in all experiments, while fluorescein isothiocyanate (FITC) gave positive results in about half of them. The prohaptens eugenol and isoeugenol induced significant proliferation in one out of eight and in four out of 12 experiments, respectively. Interestingly, in 16 assays T cells never proliferated in the presence of sodium lauryl sulfate (SLS)-treated DC. Thus, this in vitro model allows discrimination between strong contact sensitizers and irritants. It might be very useful, therefore, for restriction of animal experimentation.
我们先前曾报道,培养的人表皮朗格汉斯细胞(LC)对半抗原特异性T细胞进行体外初次致敏,为区分强接触性致敏剂和刺激物提供了一种替代方法(Krasteva等人,1996年;Moulon等人,1993年)。然而,这种基于LC的免疫测定受到人皮肤样本可用性的限制。在本研究中,我们使用单核细胞衍生的树突状细胞(DC)来分析自体T细胞对几种变应原的增殖反应。从健康供体的外周血中纯化单核细胞,并在GM/CSF和IL-4存在下培养6 - 8天,然后在GM/CSF和TNFα存在下培养2天。通过HLA-DR、CD80、CD83和CD86抗原的强表达评估,所得细胞表现出成熟DC的表型。我们表明,在所有实验中,经三硝基苯基(TNP)处理的成熟DC诱导了显著的T细胞增殖反应,而异硫氰酸荧光素(FITC)在大约一半的实验中给出了阳性结果。前半抗原丁香酚和异丁香酚分别在八分之一和十二分之四的实验中诱导了显著增殖。有趣的是,在16次测定中,T细胞在经十二烷基硫酸钠(SLS)处理的DC存在下从未增殖。因此,这种体外模型能够区分强接触性致敏剂和刺激物。因此,它对于限制动物实验可能非常有用。