Sorrell J M, Caterson B, Caplan A I, Davis B, Schafer I A
Biology Department, Case Western Reserve University, Cleveland, Ohio.
J Invest Dermatol. 1990 Sep;95(3):347-52. doi: 10.1111/1523-1747.ep12485110.
Monoclonal antibodies that recognize carbohydrate epitopes found in keratan sulfate glycosaminoglycan chains identified both intracytoplasmic and cell-surface carbohydrates of human keratinocytes. These carbohydrates were detected, using indirect immunoperoxidase methods, both in sections of paraffin-embedded tissues and in intact cultured keratinocytes. Of the seven anti-keratan sulfate monoclonal antibodies used in this study, five detected significant amounts of epitopes associated with keratinocytes. This indicates that only certain, specific types of keratan sulfate-like carbohydrates were expressed by these cells. The extent and localization of keratan sulfate-like carbohydrates appeared to be closely related to the differentiation status of cultured keratinocytes. These epitopes were very weakly expressed on surfaces of all monolayer keratinocytes, but flattened, suprabasal cells in high Ca++ cultures strongly expressed keratan sulfate-like carbohydrates on their surfaces. A much larger population of cultured keratinocytes expressed intracellular keratan sulfate-like carbohydrates identified by the same five antibodies that detected surface epitopes. In monolayer cells, keratan sulfate-like carbohydrates were predominantly found in a broad perinuclear zone. In addition, three of the five immunoreactive antibodies detected epitopes that appeared at cell boundaries, specifically at sites of close cell-to-cell contact. Thus, molecules bearing carbohydrates recognized by anti-keratan sulfate antibodies appear at developmentally important stages of keratinocyte differentiation, indicating that these carbohydrates may serve as markers for molecules important in the differentiation of human keratinocytes.
识别硫酸角质素糖胺聚糖链中碳水化合物表位的单克隆抗体鉴定出了人角质形成细胞的胞浆内和细胞表面碳水化合物。使用间接免疫过氧化物酶方法,在石蜡包埋组织切片和完整培养的角质形成细胞中均检测到了这些碳水化合物。在本研究中使用的七种抗硫酸角质素单克隆抗体中,有五种检测到了与角质形成细胞相关的大量表位。这表明这些细胞仅表达某些特定类型的硫酸角质素样碳水化合物。硫酸角质素样碳水化合物的程度和定位似乎与培养的角质形成细胞的分化状态密切相关。这些表位在所有单层角质形成细胞表面表达非常弱,但在高钙培养条件下扁平的基底上层细胞在其表面强烈表达硫酸角质素样碳水化合物。通过检测表面表位的相同五种抗体鉴定出,大量培养的角质形成细胞表达细胞内硫酸角质素样碳水化合物。在单层细胞中,硫酸角质素样碳水化合物主要存在于宽广的核周区域。此外,五种免疫反应性抗体中的三种检测到了出现在细胞边界的表位,特别是在细胞间紧密接触的部位。因此,带有抗硫酸角质素抗体识别的碳水化合物的分子出现在角质形成细胞分化的重要发育阶段,表明这些碳水化合物可能作为人类角质形成细胞分化中重要分子的标志物。