Nagatsuka Y, Watarai S, Yasuda T, Higashi H, Yamagata T, Ono Y
Department of Microbiology, Nihon University School of Medicine, Tokyo, Japan.
Immunol Lett. 1995 May;46(1-2):93-100. doi: 10.1016/0165-2478(95)00028-4.
To study the differentiation-associated glycolipid two anti-i mAb producers, GL-1 and GL-2, were established from the combination of EBV-induced transformation of normal PBL and immune lysis of fluorescent dye-trapped liposome-containing bovine i-active glycolipid. The mAb GL-1 reacted with both sialosylparagloboside and pentahexosyl ceramide and the bovine i-active glycolipid whereas mAb GL-2 reacted only with the bovine i-active glycolipid in LILA. Both mAbs cold-agglutinate human cord red cells but not adult red cells. However, unexpectedly, the majority of the reactivity of these mAbs in human cord red cells on TLC was not identical to the i-active glycolipid. The GL-1 antigenic substance is considered to be a glycolipid distinct from the i-active glycolipid because the immunoreactivity was canceled with endoglycoceramidase which cleaves a linkage between the oligosaccharide and ceramide. Based on complement cytolysis with the mAb, 15 hematopoietic cell lines and normal peripheral lymphocytes were screened for susceptibility to the mAbs. A Burkitt lymphoma cell line, Ramos, was most sensitive among those tested, and BJA-B, Daudi, Namalwa in the B cell lines, TALL-1, Jurkatt in the T-cell lines and HL-60 in the non-lymphoid cell lines were sensitive whereas normal lymphocytes or other 8 cell lines were not. An immunoreactive spot with the same Rf with cord red cells was also detected in sensitive cell lines. The possible presence of a new glycolipid antigen determined from the mAb and related to the differentiation of hematopoietic cells was speculated.
为研究分化相关糖脂，从EB病毒诱导的正常外周血淋巴细胞转化与含牛i活性糖脂的荧光染料捕获脂质体免疫裂解相结合的过程中建立了两种抗i单克隆抗体产生细胞系GL-1和GL-2。单克隆抗体GL-1与唾液酸副球蛋白、五己糖神经酰胺以及牛i活性糖脂发生反应，而单克隆抗体GL-2在LILA中仅与牛i活性糖脂发生反应。两种单克隆抗体均能冷凝聚人脐带红细胞，但不能凝聚成人红细胞。然而，出乎意料的是，这些单克隆抗体在人脐带红细胞薄层色谱上的大部分反应性与i活性糖脂并不相同。GL-1抗原物质被认为是一种与i活性糖脂不同的糖脂，因为其免疫反应性可被内切糖苷酶消除，该酶可切断寡糖与神经酰胺之间的连接。基于单克隆抗体的补体细胞溶解作用，对15种造血细胞系和正常外周淋巴细胞进行了对单克隆抗体敏感性的筛选。在所测试的细胞系中，伯基特淋巴瘤细胞系Ramos最为敏感，B细胞系中的BJA-B、Daudi、Namalwa，T细胞系中的TALL-1、Jurkatt以及非淋巴细胞系中的HL-60均敏感，而正常淋巴细胞或其他8种细胞系则不敏感。在敏感细胞系中也检测到了与脐带红细胞具有相同比移值的免疫反应斑点。推测可能存在一种由单克隆抗体确定的、与造血细胞分化相关的新糖脂抗原。
