Bessou S, Gauthier Y, Surlève-Bazeille J E, Pain C, Taïeb A
Laboratoire de Dermatologie, Université Victor Ségalen, Bordeaux II, France.
Br J Dermatol. 1997 Dec;137(6):890-7.
The primary cellular or molecular targets accounting for melanocytes loss in vitiligo are not clearly identified. To study a putative latent epidermal defect in the epidermis of vitiligo patients, we performed in vitro studies using cultured vitiligo melanocytes and keratinocytes transplanted on to a dead de-epidermized dermis according to a variant of Pruniéras' technique. Control autologous constructs were made with keratinocytes and melanocytes of normal adult epidermis and vitiligo epidermis from perilesional skin. For heterologous reconstructs we combined vitiligo-derived melanocytes or keratinocytes with their normal phototype-matched counterpart. After 15 days of culture at the air-liquid interface, epidermal reconstructs were studied macroscopically and microscopically. Immunohistochemistry was performed using antibodies to TRP-1 and NKI-beteb. All heterologous and autologous reconstructs made with melanocytes and keratinocytes from vitiligo patients had a normal histology and ultrastructure. For vitiligo melanocytes or normal melanocytes submitted to the influence of vitiligo keratinocytes, immunophenotype and function (pigment production and transfer) were similar to normal controls. So, without additional noxious stimuli, we could not discriminate between melanocytes and keratinocytes as inducers of the disease. Our data suggest that the basic abnormality in vitiligo vulgaris needs extrinsic factors to be macroscopically revealed or requires a longer period of culture to develop. Our model will allow analysis of the various pathophysiological mechanisms of vitiligo, e.g. autoantibodies or oxidative stress, at the cellular, biochemical or molecular level.
白癜风中导致黑素细胞缺失的主要细胞或分子靶点尚未明确。为了研究白癜风患者表皮中一种假定的潜在缺陷，我们采用Pruniéras技术的变体，将培养的白癜风黑素细胞和角质形成细胞移植到死亡的去表皮真皮上，进行了体外研究。用正常成人表皮以及白癜风患者皮损周围皮肤的角质形成细胞和黑素细胞制作对照自体构建体。对于异种重建，我们将白癜风来源的黑素细胞或角质形成细胞与其正常光型匹配的对应细胞相结合。在气液界面培养15天后，对表皮重建体进行宏观和微观研究。使用针对TRP-1和NKI-beteb的抗体进行免疫组织化学分析。所有用白癜风患者的黑素细胞和角质形成细胞制作的异种和自体重建体均具有正常的组织学和超微结构。对于受到白癜风角质形成细胞影响的白癜风黑素细胞或正常黑素细胞，其免疫表型和功能（色素产生和转移）与正常对照相似。因此，在没有额外有害刺激的情况下，我们无法区分黑素细胞和角质形成细胞谁是该疾病的诱导因素。我们的数据表明，寻常型白癜风的基本异常需要外在因素才能在宏观上显现出来，或者需要更长的培养时间才会发展。我们的模型将允许在细胞、生化或分子水平上分析白癜风的各种病理生理机制，例如自身抗体或氧化应激。
