Monoclonal antibodies to normal and abnormal epithelial antigens.
Monoclonal antibodies to normal and abnormal epithelial antigens.
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针对正常和异常上皮抗原的单克隆抗体。
DOI：
10.1159/000408660
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Briggaman,RA
中科院分区：
文献类型：
--
作者：
Goldsmith,LA;Briggaman,RA
用ME-180（一种人宫颈癌细胞系）免疫Balb/c小鼠，形成了抗各种人表皮和基底膜成分的小鼠单克隆抗体。使用聚乙二醇将来自超免疫小鼠的脾细胞与非分泌型小鼠骨髓瘤细胞系融合。通过在24孔Linbro板中的RPMI-1640中含有20%胎牛血清、次黄嘌呤、胸苷和甲氨蝶呤的培养基中生长来选择所得杂交体。使产生感兴趣的抗体的威尔斯孔生长并最终克隆到HGPRT大鼠成纤维细胞饲养层上。两个克隆的目的抗体是DUX 5.2和DUX 1.1。3. DUX 5. 2是小鼠IgG的一个亚类，与ME-180细胞膜和人皮肤表皮基底膜区反应，如直接免疫荧光显微镜所示。使用电子显微镜免疫过氧化物酶技术的超微结构定位显示DUX 5.2抗原定位在致密层下方;反应产物可能包括锚定原纤维。尽管DUX 5.2与正常人基底膜区和几种疾病中的基底膜区反应，但在营养不良性大疱性表皮病（DEB）患者的正常、永不发红的皮肤中没有反应性。这表明疾病中胶原酶的增加可能会破坏DUX 5.2识别的抗原的抗原性，或者抗原可能不存在于DEB中。因此，该抗体将允许DEB的早期新生儿和产前诊断，并允许分离缺乏的结构部分
Mouse monoclonal antibodies to various human epidermal and basement membrane components were formed by immunizing Balb/c mice with ME-180, a line of human cervical carcinoma cells. The spleen cells from hyperimmunized mice were fused with a nonsecreting mouse myeloma cell line using polyethylene glycol. The resulting hybrids were selected by growth in media containing 20% fetal calf serum, hypoxan-thine, thymidine, and methotrexate in RPMI-1640 in 24-well Linbro plates. Wells producing antibodies of interest were grown and eventually cloned over an HGPRT rat fibroblast feeder layer. These cultures were expanded and recloned.Two cloned antibodies of interest are DUX 5.2 and DUX 1.1. 3. DUX 5. 2 is the mouse IgG₁ subclass and reacts with the membranes of ME-180 cells and the human skin epidermal basement membrane zone as shown by direct immunofluorescent microscopy. Ultrastructural localization using electron microscopic immunoperoxidase techniques showed localization of the DUX 5.2 antigen to be beneath the lamina densa; the reaction product may include the anchoring fibrils. Although DUX 5.2 reacts with the normal human basement membrane zone and the basement membrane zone in several diseases, there is no reactivity in the normal, never-blistered skin of patients with dystrophic epidermolysis bullosa (DEB). This suggests that the increased collagenase in the disease may be destroying antigenicity of the antigen recognized by DUX 5.2 or that the antigen may not be present in DEB. This antibody will thus allow early neonatal and prenatal diagnosis in DEB and allow isolation of the structural moiety which is deficient in