Loveless R W, Yuen C T, Tsuiji H, Irimura T, Feizi T
The Glycosciences Laboratory, Imperial College School of Medicine, Northwick Park Hospital, Harrow, Middlesex, HA1 3UJ, United Kingdom.
Glycobiology. 1998 Dec;8(12):1237-42. doi: 10.1093/glycob/8.12.1237.
The IgG1hybridoma antibody, 91.9H, was originally raised against sulfated mucins isolated from normal human colonic mucosa. Previous studies have shown that the 91.9H antigen is expressed on normal colonic epithelial cells and the sulfomucins that they produce, but not in the normal small intestine and stomach. Tissue-specific changes occur in 91.9H antigen expression in disease: the antigen diminishes in colonic carcinomas, whereas in regions of gastric mucosa showing intestinal metaplasia and in gastric carcinomas, the antigen is expressed as a "neo-antigen." This report is concerned with elucidation, by the neoglycolipid technology, of the determinant recognized by antibody 91.9H using sulfated and sialyl oligosaccharides of Lewisa(Lea) and Lextypes, and analogs that lack sulfate, sialic acid, or fucose. Binding experiments with the lipid-linked oligosaccharides immobilized on chromatograms or on microwells, and inhibition of binding experiments with free oligosaccharides based on di-, tri- and tetrasaccharide backbones, show that the 91.9H antigenic determinant is based on a trisaccharide backbone, and consists of the 3'-sulfated Leatetrasaccharide sequence, which is a potent ligand for the E- and L-selectins. The antibody gives a relatively low signal with the 3'-sulfated non-fucosylated backbone, and has no detectable cross-reaction with the 3'-sulfated Lexisomer, nor with sialyl-Leaand -Lexanalogues. Antibody 91.9H is a valuable addition, therefore, to the repertoire of reagents for mapping details of the distribution, and determining the relative importance of sulfated and sialyl oligosaccharides as ligands for the selectins, in normal and pathological epithelia and endothelia.
IgG1杂交瘤抗体91.9H最初是针对从正常人结肠黏膜分离出的硫酸化黏蛋白产生的。先前的研究表明，91.9H抗原在正常结肠上皮细胞及其产生的硫酸黏蛋白上表达，但在正常小肠和胃中不表达。在疾病中，91.9H抗原表达会出现组织特异性变化：该抗原在结肠癌中减少，而在显示肠化生的胃黏膜区域和胃癌中，该抗原作为“新抗原”表达。本报告旨在通过新糖脂技术，利用Lewis a（Lea）和Lex型的硫酸化和唾液酸化寡糖以及缺乏硫酸、唾液酸或岩藻糖的类似物，阐明抗体91.9H识别的决定簇。用固定在色谱图或微孔上的脂质连接寡糖进行结合实验，以及用基于二糖、三糖和四糖主链的游离寡糖进行结合抑制实验，结果表明91.9H抗原决定簇基于三糖主链，由3'-硫酸化Lea四糖序列组成，该序列是E-和L-选择素的有效配体。该抗体与3'-硫酸化非岩藻糖基化主链的信号相对较低，与3'-硫酸化Lex异构体以及唾液酸化-Lea和-Lex类似物均无明显交叉反应。因此，抗体91.9H是一种有价值的试剂，可用于绘制正常和病理上皮及内皮中分布细节图谱，并确定硫酸化和唾液酸化寡糖作为选择素配体的相对重要性。
