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。得到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。另外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都能识别碳水化合物部分。Western blotting结果显示，大多数抗体识别的是高分子量物质，但有两个抗原经过SDS和热处理变性。还测定了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。对慢性鼻窦炎和变应性鼻炎患者的鼻分泌物进行点印迹分析，观察抗体与这些抗体的不同反应性。鼻上皮的免疫组织学研究也揭示了慢性鼻窦炎、变应性鼻炎和术后上颌囊肿患者鼻分泌物的疾病特异性变化。我们开发了一种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对该培养系统中粘液分泌量的量化非常有用。

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.