Sasaki K, Okouchi Y, Rothkötter H J, Pabst R
Department of Anatomy, Yamagata University School of Medicine, Japan.
Anat Rec. 1996 Jan;244(1):105-11. doi: 10.1002/(SICI)1097-0185(199601)244:1<105::AID-AR10>3.0.CO;2-T.
The high endothelial venules (HEV) in the lymph nodes are essential for lymphocyte recirculation. As a first step, the HEV surface interacts with lymphocytes through adhesion molecules. It is important to know where adhesion molecules are expressed on the surface ultrastructure and how these structures interact with lymphocytes.
To demonstrate the ultrastructural mechanism of interaction between the HEV surface and lymphocytes through the intercellular adhesion molecule (ICAM-1), rat mesenteric lymph nodes were perfused through the superior mesenteric artery with the primary antibody (antirat ICAM-1 antibody) and secondary antibody (antimouse IgG coupled to 15 nm gold particles), which were diluted with hypothermic University of Wisconsin (UW) solution. After the immunoreaction, we analyzed the HEV three-dimensionally and quantitatively using immunoscanning electron microscopy (ISEM) combined with transmission electron microscopy (TEM).
HEV expressed ICAM-1 in a 5-30-fold higher concentration than other vessels. Its distribution was extensive over the luminal surface of the cell down to the junctional area. The endothelial surface of HEV undulated to form branched microfolds, along which ICAM-1 was expressed. Cytoplasmic processes of lymphocytes were seen in microfurrows between microfolds and adhered to the sides of the folds.
These observations imply that the microfolds expressing ICAM-1 and microfurrows are specific ultrastructural features for trapping lymphocytes, thus initiating lymphocyte emigration into the lymph node parenchyma.
淋巴结中的高内皮微静脉(HEV)对于淋巴细胞再循环至关重要。第一步,HEV表面通过黏附分子与淋巴细胞相互作用。了解黏附分子在表面超微结构上的表达位置以及这些结构如何与淋巴细胞相互作用非常重要。
为了通过细胞间黏附分子(ICAM-1)证明HEV表面与淋巴细胞相互作用的超微结构机制,用一级抗体(抗大鼠ICAM-1抗体)和二级抗体(与15纳米金颗粒偶联的抗小鼠IgG)经肠系膜上动脉灌注大鼠肠系膜淋巴结,抗体用低温的威斯康星大学(UW)溶液稀释。免疫反应后,我们结合透射电子显微镜(TEM)使用免疫扫描电子显微镜(ISEM)对HEV进行三维定量分析。
HEV表达ICAM-1的浓度比其他血管高5至30倍。其分布在细胞腔表面直至连接区域广泛存在。HEV的内皮表面起伏形成分支微褶,ICAM-1沿微褶表达。在微褶之间的微沟中可见淋巴细胞的细胞质突起,并附着在褶的侧面。
这些观察结果表明,表达ICAM-1的微褶和微沟是捕获淋巴细胞的特定超微结构特征,从而启动淋巴细胞向淋巴结实质的迁移。