Qureshi R, Forstner G G, Forstner J F
J Clin Invest. 1979 Nov;64(5):1149-56. doi: 10.1172/JCI109568.
We have developed a double-antibody radioimmunoassay for the quantitative measurement of human goblet cell mucin (GCM) in order to study intestinal mucus in human and other species. The assay used 3H-labeled mucin as the antigen, rabbit antisera, and sheep anti-rabbit IgG antisera as the second antibody. A number of applications of the assay were investigated. A survey of human tissues revealed that mucins of the rectum, colon, and small intestine had identical affinity for the rabbit antibody, whereas lung eyelid conjunctiva, esophagus, and stomach reacted less strongly. GCM concentration ranged from 1.9 to 14 microgram mucin protein/mg tissue protein in the small and large intestine, respectively. The radioimmunoassay was also found to be useful as a marker during the isolation of GCM from human ileal extracts, where it indicated that a 10,000-fold purification had been achieved. Antigenic determinants of the mucin did not rely upon ABH blood group-specific terminal sugars in oligosaccharide chains. A comparison of mucins among various species revealed a partial species specificity of the GCM antibody. Human GCM cross-reacted with dog, monkey, and rabbit mucins, but not with mucins of rat, pig, toad, and oyster. Organ distributions of cross-reactive mucins in rabbit tissues indicated a pattern that was qualitatively similar to that seen in human tissues. Possible implications of these findings for autoimmune diseases are briefly discussed.
为了研究人类和其他物种的肠道黏液,我们开发了一种用于定量检测人杯状细胞黏蛋白(GCM)的双抗体放射免疫分析法。该分析方法使用³H标记的黏蛋白作为抗原,兔抗血清,以及羊抗兔IgG抗血清作为第二抗体。我们研究了该分析方法的多种应用。对人体组织的一项调查显示,直肠、结肠和小肠的黏蛋白对兔抗体具有相同的亲和力,而肺、眼睑结膜、食管和胃的反应则较弱。在小肠和大肠中,GCM浓度分别为1.9至14微克黏蛋白蛋白/毫克组织蛋白。放射免疫分析法在从人回肠提取物中分离GCM的过程中也被证明是一种有用的标记物,它表明已经实现了10000倍的纯化。黏蛋白的抗原决定簇并不依赖于寡糖链中ABH血型特异性末端糖。对不同物种黏蛋白的比较揭示了GCM抗体的部分物种特异性。人GCM与狗、猴和兔的黏蛋白发生交叉反应,但与大鼠、猪、蟾蜍和牡蛎的黏蛋白不发生交叉反应。兔组织中交叉反应黏蛋白的器官分布表明其模式在质量上与人类组织中观察到的相似。本文简要讨论了这些发现对自身免疫性疾病的可能影响。