Mizukawa Y, Shitara K, Yamazaki Y, Kudo A, Narimatsu H, Shiohara T
Department of Dermatology, Kyorin University School of Medicine, Mitaka, Tokyo, Japan.
Lab Invest. 2001 May;81(5):771-3. doi: 10.1038/labinvest.3780286.
Although expression of cutaneous lymphocyte-associated antigen (CLA) is thought to be a specific marker of skin "homing" T cells, it has become clear that CLA is a good marker for high levels of fucosyltransferase VII (Fuc-TVII) activity but does not necessarily represent the epitope required for binding to E-selectin (Wagers et al, 1996, 1997). Therefore, expression of Fuc-TVII is an attractive candidate for identifying skin "homing" T cells. However, analyses of Fuc-TVII expression in human T cells have been performed only at the mRNA level (Nakayama et al, 2000) because of the lack of mAB: In this study Fuc-TVII was for the first time visualized in individual cells by using a novel mAb that we developed. Double immunofluorescence and immunohistochemistry demonstrated the coexpression of Fuc-TVII and CLA in cell lines in which Fuc-TVII mRNA was shown to be expressed at high levels: whereas CLA expression was seen in the cell membrane, Fuc-TVII was identified in a supra- or perinuclear location. Cytoplasmic Fuc-TVII expression was also detectable in both CD4(+) and CD8(+) T cells purified from peripheral blood. Fuc-TVII was also expressed at high levels in many CLA(+) T cells infiltrating the skin. In these peripheral T cells, unlike in cell lines, cytoplasmic expression of Fuc-TVII was not always associated with surface CLA expression. This mAb would serve as a valuable tool for selectively identifying a novel subset of skin "homing" T cells that are not detected by the conventional method because of the lack of CLA expression.
尽管皮肤淋巴细胞相关抗原(CLA)的表达被认为是皮肤“归巢”T细胞的特异性标志物,但现在已经明确,CLA是岩藻糖基转移酶VII(Fuc-TVII)高活性的良好标志物,但不一定代表与E-选择素结合所需的表位(Wagers等人,1996年,1997年)。因此,Fuc-TVII的表达是鉴定皮肤“归巢”T细胞的一个有吸引力的候选指标。然而,由于缺乏单克隆抗体,对人T细胞中Fuc-TVII表达的分析仅在mRNA水平上进行(Nakayama等人,2000年):在本研究中,我们首次使用我们开发的新型单克隆抗体在单个细胞中可视化Fuc-TVII。双重免疫荧光和免疫组织化学证明,在Fuc-TVII mRNA被证明高水平表达之细胞系中,Fuc-TVII和CLA共表达:虽然CLA表达见于细胞膜,但Fuc-TVII定位于核上或核周位置。从外周血纯化的CD4(+)和CD8(+) T细胞中也可检测到细胞质Fuc-TVII表达。在许多浸润皮肤的CLA(+) T细胞中,Fuc-TVII也高水平表达。在这些外周T细胞中,与细胞系不同,Fuc-TVII的细胞质表达并不总是与表面CLA表达相关。这种单克隆抗体将成为一种有价值的工具,用于选择性鉴定因缺乏CLA表达而无法用传统方法检测到的新型皮肤“归巢”T细胞亚群。