Identification of a major carbohydrate capping group of the L-selectin ligand on high endothelial venules in human lymph nodes as 6-sulfo sialyl Lewis X
Identification of a major carbohydrate capping group of the L-selectin ligand on high endothelial venules in human lymph nodes as 6-sulfo sialyl Lewis X
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DOI：
10.1074/jbc.273.18.11225
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发表时间：
1998-05-01
影响因子：
4.8
通讯作者：
Kannagi, R
中科院分区：
文献类型：
--
作者：
Mitsuoka, C;Sawada-Kasugai, M;Kannagi, R
我们研究了硫酸化唾液酸路易斯 X 决定簇（假定的 L-选择素配体）的分子种类，在人类淋巴结的高内皮小静脉 (HEV) 上表达。将 HEV 的反应模式与纯 6-磺基、6'-磺基或 6,6'-双磺基唾液酸路易斯 X 决定簇与迄今为止已知的抗唾液酸路易斯 X 抗体的反应性进行比较，强烈表明 6-磺基唾液酸 Leu 是 X 是 HEV 上主要硫酸化唾液酸路易斯 X 决定簇的最佳候选，其次是 6,6'-双磺基唾液酸路易斯 X 决定簇X，而 6'-磺基唾液酸路易斯 X 不太可能。我们新生成了针对 B-磺基唾液酸 Lewis X 的单克隆抗体 (mAb) G152 和 G72，它们在免疫组织化学检查中强烈标记 HEV，并抑制重组 L-选择素-IgG 与 HEV 的结合，表明该决定簇充当 L-选择素的配体。为了测试 6,6'-bissulfo sialyl Lewis X 的伴随表达，生成了特异性 mAb（G2706、G27011、G27037 和 G27039），但所有抗体均未能与 HEV 发生反应，接下来，我们建立了针对 B-sulfo Lewis X（B-sulfo sialyl Lewis X 的去唾液酸形式）的 mAb（AG97 和 AG273）。对未经处理的 HEV 没有反应，但对唾液酸酶处理的 HEV 反应强烈，这表明 6-磺基唾液酸 Lewis X 的唾液酸化形式占主导地位，其脱唾液酸形式表达最少，证实它是由岩藻糖基转移酶 VII 合成的，岩藻糖基转移酶 VII 是优先产生决定簇唾液酸化形式的同工酶。
We investigated the molecular species of sulfated sialyl Lewis X determinants, the putative L-selectin ligand, expressed on high endothelial venules (HEV) in human lymph nodes. Comparison of the reactivity pattern of HEV with the reactivity of the pure 6-sulfo, 6'-sulfo, or 6,6'-bissulfo sialyl Lewis X determinant with hitherto known anti-sialyl Lewis X antibodies strongly suggested 6-sulfo sialyl Leu is X to be the best candidate for the major sulfated sialyl Lewis X determinant on HEV, followed by 6,6'-bissulfo sialyl Lewis X, whereas 6'-sulfo sialyl Lewis X was unlikely. We newly generated mono clonal antibodies (mAbs) G152 and G72 directed against B-sulfo sialyl Lewis X, which intensely labeled HEV in immunohistochemical examination and inhibited binding of recombinant L-selectin-IgG to HEV, suggesting that the determinant serves as a ligand for L-selectin. To test the concomitant expression of 6,6'-bissulfo sialyl Lewis X, specific mAbs (G2706, G27011, G27037, and G27039) were generated, but all antibodies failed to react to HEV, Next, we established mAbs (AG97 and AG273) directed against B-sulfo Lewis X, the asialo form of B-sulfo sialyl Lewis X, The antibodies were not reactive to untreated HEV, but strongly reacted to sialidase-treated HEV, This indicated the predominance of the sialylated form of 6-sulfo sialyl Lewis X and minimal expression of its asialo form, corroborating that it was synthesized by fucosyltransferase VII, the isoenzyme that preferentially produces the sialylated form of the determinant.