BINDING OF ANTIBODIES TO THE EXTRACTABLE NUCLEAR ANTIGENS SS-A/RO AND SS-B/LA IS INDUCED ON THE SURFACE OF HUMAN KERATINOCYTES BY ULTRAVIOLET-LIGHT (UVL) - IMPLICATIONS FOR THE PATHOGENESIS OF PHOTOSENSITIVE CUTANEOUS LUPUS
BINDING OF ANTIBODIES TO THE EXTRACTABLE NUCLEAR ANTIGENS SS-A/RO AND SS-B/LA IS INDUCED ON THE SURFACE OF HUMAN KERATINOCYTES BY ULTRAVIOLET-LIGHT (UVL) - IMPLICATIONS FOR THE PATHOGENESIS OF PHOTOSENSITIVE CUTANEOUS LUPUS
复制标题
DOI：
10.1111/1523-1747.ep12873930
复制
发表时间：
1990-01-01
影响因子：
6.5
通讯作者：
NORRIS, DA
中科院分区：
文献类型：
--
作者：
FURUKAWA, F;KASHIHARASAWAMI, M;NORRIS, DA
非组蛋白核蛋白抗原SS-A/Ro、SS-B/La和RNP的自身抗体与光敏性皮肤红斑狼疮（LE）高度相关。为了更好地了解紫外线（UV）光对皮肤LE患者光敏性的潜在机制，我们设计了体外和体内免疫病理学实验，以评估UV对此类自身抗体与人角质形成细胞表面结合的影响，这是光敏LE免疫损伤的主要靶点之一。短期的2%多聚甲醛固定的悬浮液培养的人角质形成细胞先前孵育的单特异性抗血清探针，使ENA表达的细胞表面上的流式细胞术分析检测。UVB光（280-320 nm）以剂量依赖性模式诱导角质形成细胞上SS-A/Ro和SS-B/La的单特异性抗体探针的结合，在200 mJ/cm 2 UVB剂量下观察到最大诱导。SS-A/Ro、SS-B/La和RNP抗体的结合强烈增强，但抗Sm的结合很弱。相反，UVA（320-400 nm）光对这些抗体探针的结合的诱导没有影响。通过标准免疫荧光技术观察到相同的结果。羟基脲处理的角质形成细胞显示出类似的诱导这些抗原的UVB照射，这表明ENA的表达由UVB培养的角质形成细胞的细胞周期无关。蛋白质糖基化抑制剂衣霉素（Tunicamycin）可降低UVB对SS-A/Ro和SS-B/La抗原表达的影响。这些体外FACS分析显示，ENA对角质形成细胞表面的增强是剂量依赖性的、UVB依赖性的、糖基化依赖性的和细胞周期无关的。在体内ENA增加角质形成细胞表面上进行了检查，在吸水泡表皮屋顶。SS-A/Ro，SS-B/La，RNP和Sm的特异性抗体探针结合到体内UVL照射后完整抽吸水疱表皮中的人角质形成细胞。使用三种不同的方案，我们已经证明，抗体SS-A/Ro，SS-B.La，和U1 RNP结合紫外线照射的人角质形成细胞。我们推测这种抗体结合是光敏性皮肤狼疮中抗体依赖性角质形成细胞损伤的重要诱导物。
Autoantibodies to the non-histone nucleoprotein antigens SS-A/Ro, SS-B/La, and RNP are highly associated with photosensitive cutaneous lupus erythematosus (LE). In order to better understand the potential mechanisms of ultraviolet (UV) light on photosensitivity in patients with cutaneous LE, we designed immunopathologic in vitro and in vivo experiments to evaluate the effects of UV on the binding of such autoantibodies to the surface of human keratinocytes, one major target of immunologic damage in photosensitive LE. Short-term 2% paraformaldehyde fixation of suspensions of cultured human keratinocytes previously incubated with monospecific antiserum probes enabled the detection of ENA expression on the cell surface by flow-cytometry analysis. UVB light (280-320 nm) induced the binding of monospecific antibody probes for SS-A/Ro and SS-B/La on keratinocytes in a dose-dependent pattern with maximal induction observed at the dose of 200 mJ/cm2 UVB. Binding of SS-A/Ro, SS-B/La, and RNP antibody was augmented strongly, but binding of anti-Sm was very weak. In contrast, UVA (320-400 nm) light had no effect on the induction of binding of these antibody probes. Identical results were seen by standard immunofluorescence techniques. Hydroxyurea-treated keratinocytes showed similar induction of those antigens by UVB irradiation, which suggested that ENA expression on cultured keratinocytes by UVB were cell-cycle independent. Tunicamycin, an inhibitor of glycosylation of proteins, reduced UVB light effect on the SS-A/Ro and SS-B/La antigen''s expression. These in vitro FACS analyses revealed that ENA augmentation on the keratinocyte cell surface was dose dependent, UVB dependent, glycosylation dependent, and cell-cycle independent. In vivo ENA augmentation on the keratinocyte surface was examined in suction blister epidermal roofs. Specific antibody probes for SS-A/Ro, SS-B/La, RNP, and Sm bound to human keratinocytes in intact suction blister epidermis following UVL irradiation in vivo. Using three different protocols, we have demonstrated that antibodies to SS-A/Ro, SS-B.La, and U1RNP bind to UVL-irradiated human keratinocytes. We speculate that this antibody binding is an important inducer of antibody dependent keratinocyte damage in photosensitive cutaneous lupus.