Mitsuoka C, Kawakami-Kimura N, Kasugai-Sawada M, Hiraiwa N, Toda K, Ishida H, Kiso M, Hasegawa A, Kannagi R
Program of Experimental Pathology, Aichi Cancer Center, Research Institute, Nagoya, Japan.
Biochem Biophys Res Commun. 1997 Jan 23;230(3):546-51. doi: 10.1006/bbrc.1996.6012.
Endothelial cells of high endothelial venules (HEV) in human peripheral lymph nodes expressed a distinct type of sialyl Lewis X antigen, which was detected preferentially with a set of anti-sialyl Lewis X antibodies, 2F3, 2H5 and HECA-452 in immunohistochemistry, while another set of anti-sialyl Lewis X antibodies, FH-6 and CSLEX-1, failed to detect it. The adhesion of cells expressing L-selectin to HEV was inhibited by members of the former set of antibodies in Stamper-Woodruff assays performed on frozen sections of human peripheral lymph nodes. Transfection of a cultured endothelial cell line with a human alpha1-->3 fucosyltransferase, Fuc-T VII, resulted in the expression of a distinct type of sialyl Lewis X antigen having the reactivity similar to that of HEV; i.e., the antigen appearing on the transfectant clone was detectable only with the set of 2F3, 2H5 and HECA-452, but not with the set of FH-6 and CSLEX-1. Treatment of transfectant cells with sodium chlorate, a metabolic inhibitor of sulfation, resulted in reactivity to the members of the latter set of antibodies, suggesting that sulfation of sialyl Lewis X moiety was the cause of the discrepancy in the reactivity of the anti-sialyl Lewis X antibodies. When tested against various authentic sulfated sialyl Lewis X determinants, 6-sulfo sialyl Lewis X and 6,6'-bis-sulfo sialyl Lewis X were found to be reactive to the antibodies, 2F3, 2H5 and HECA-452, but not with antibodies FH-6 and CSLEX-1, suggesting that the distinct type of sialyl Lewis X determinant on the HEV endothelial cells and Fuc-T VII-transfected endothelial cell clone are mainly 6-sulfo and/or 6,6'-bis-sulfo sialyl Lewis X determinants.
人外周淋巴结的高内皮微静脉（HEV）内皮细胞表达一种独特类型的唾液酸化路易斯X抗原，在免疫组织化学中，用一组抗唾液酸化路易斯X抗体2F3、2H5和HECA-452能优先检测到该抗原，而另一组抗唾液酸化路易斯X抗体FH-6和CSLEX-1则无法检测到。在对人外周淋巴结冰冻切片进行的斯坦珀-伍德拉夫试验中，前一组抗体的成员可抑制表达L-选择素的细胞与HEV的黏附。用人类α1→3岩藻糖基转移酶Fuc-T VII转染培养的内皮细胞系，导致表达一种与HEV具有相似反应性的独特类型的唾液酸化路易斯X抗原；即，转染克隆上出现的抗原仅能用2F3、2H5和HECA-452这组抗体检测到，而不能用FH-6和CSLEX-1这组抗体检测到。用硫酸化代谢抑制剂氯酸钠处理转染细胞，导致其对后一组抗体有反应，这表明唾液酸化路易斯X部分的硫酸化是抗唾液酸化路易斯X抗体反应性差异的原因。当针对各种真实的硫酸化唾液酸化路易斯X决定簇进行测试时，发现6-磺基唾液酸化路易斯X和6,6'-双磺基唾液酸化路易斯X与抗体2F3、2H5和HECA-452有反应，但与抗体FH-6和CSLEX-1无反应，这表明HEV内皮细胞和Fuc-T VII转染的内皮细胞克隆上独特类型的唾液酸化路易斯X决定簇主要是6-磺基和/或6,6'-双磺基唾液酸化路易斯X决定簇。
