Monoclonal antibodies as probes for studying human nasal secretions

制备了一系列针对慢性鼻窦炎患者鼻分泌物的单克隆抗体。获得了4种不同类型的18种单克隆抗体。抗体HCS 18与上皮杯状细胞反应。抗体HCS 4、5、6和16染色粘膜下腺细胞。抗体HCS 13和15与上皮杯状细胞、粘膜下腺细胞和血管内皮细胞反应。其他11种抗体识别上皮杯状细胞和粘膜下腺细胞。抗原的高碘酸盐敏感性表明，所有抗体识别碳水化合物部分。Western blotting结果显示，大部分抗体识别高分子量物质，但有两种抗原经SDS和热处理后发生变性。这些抗体与其他器官和其他物种的分泌细胞的交叉反应性也进行了测定。在斑点印迹分析混合鼻分泌物从慢性鼻窦炎和变应性鼻炎患者观察到不同的反应性与这些抗体。免疫组化研究还发现慢性鼻窦炎、变应性鼻炎和上颌窦囊肿术后鼻分泌物的变化，我们建立了一个气液界面的体内细胞培养系统来研究粘液分泌的机制，杯状细胞特异性抗体HCS 18是非常有用的，以定量的粘液分泌量在这个培养系统中。

A series of monoclonal antibodies were generated against nasal secretions from patients with chronic sinusitis. Four different types of eighteen monoclonal antibodies were obtained. Antibody HCS 18 reacted with epithelial goblet cells. Antibody HCS 4,5,6, and 16 stained submucosal gland cells. Antibody HCS 13 and 15 reacted with epithelial goblet cells, submucosal gland cells and endothelial cells of vessels. Other eleven antibodies recognized epithelial goblet cells and submucosal gland cells. Periodate sensitivity of antigens suggested that all antibodies recognized carbohydrate moieties. Western blotting revealed that most of antibodies recognized high molecular weight materials, but two antigens were denatured by SDS and heat treatment. Cross-reactivity of these antibodies with secretory cells in other organs and in other species were also determined.The different reactivities with these antibodies were observed in dot blot analysis of pooled nasal secretions from patients with chronic sinusitis and allergic rhinitis. Immunohistological studies of nasal epithelium also revealed the disease-specific changes of nasal secretions in chronic sinusitis, allergic rhinitis, and postoperative maxillary cyst.We have developed an air-liquid interface in vivo cell culture system to study the mechanisms of mucus secretion, and goblet cell-specific antibody HCS 18 is very useful to quantify the amount of mucus secretion in this culture system.