Dustin M L, Carpen O, Springer T A
Center for Blood Research, Boston, MA 02115.
J Immunol. 1992 May 1;148(9):2654-63.
We demonstrate complementary differences in the behavior of B lymphoblastoid cells adhering to LFA-1 or its counter-receptor ICAM-1. The interaction of B lymphoblastoid cells with glass-supported planar bilayers bearing LFA-1 or ICAM-1 was observed by time-lapse video microscopy, and the distribution of adhesion receptors on cells interacting with the planar bilayers was studied by immunofluorescence microscopy. B lymphoblasts formed a large contact area and crawled rapidly (up to 25 microns/min) on planar bilayers bearing ICAM-1. In contrast, these cells attached to planar bilayers bearing LFA-1 through a fixed point about which the cells actively pivoted, using a single stalk-like projection. Phorbol ester-stimulated lymphoblasts, which adhere more strongly to ICAM-1-bearing substrates than unstimulated lymphoblasts, were still capable of locomotion on ICAM-1. Phorbol ester stimulation of B lymphoblasts on planar bilayers bearing LFA-1 promoted a rapid conversion from "stalk" attachment to symmetrical spreading of the cell on the substrate. Cellular LFA-1 remained uniformly distributed on the cell surface during interaction with bilayers bearing purified ICAM-1 as determined by immunofluorescence. In contrast, ICAM-1 was concentrated in the stalk-like structure through which the unstimulated B lymphoblasts adhered to LFA-1 in planar bilayers, but ICAM-1 immunofluorescence became more uniformly distributed over the cell surface within minutes of phorbol ester addition. Neither LFA-1 or ICAM-1 colocalized with the prominent staining of filamentous actin in the ruffling membrane regions. Interaction through cell surface LFA-1 and ICAM-1, 2, or 3 promotes different cellular morphologies and behaviors, the correlation of which with previously observed patterns of lymphocyte interaction with different cell types is discussed.
我们证明了B淋巴母细胞在黏附于淋巴细胞功能相关抗原-1(LFA-1)或其反受体细胞间黏附分子-1(ICAM-1)时表现出互补性差异。通过延时视频显微镜观察B淋巴母细胞与负载LFA-1或ICAM-1的玻璃支撑平面双层膜的相互作用,并通过免疫荧光显微镜研究与平面双层膜相互作用的细胞上黏附受体的分布。B淋巴母细胞在负载ICAM-1的平面双层膜上形成大的接触面积并快速爬行(速度可达25微米/分钟)。相比之下,这些细胞通过一个固定点附着在负载LFA-1的平面双层膜上,细胞围绕该点积极旋转,使用单个茎状突起。佛波酯刺激的淋巴母细胞比未刺激的淋巴母细胞更强烈地黏附于负载ICAM-1的底物,但仍能够在ICAM-1上移动。在负载LFA-1的平面双层膜上对B淋巴母细胞进行佛波酯刺激促进了从“茎”附着到细胞在底物上对称铺展的快速转变。通过免疫荧光测定,在与负载纯化ICAM-1的双层膜相互作用期间,细胞LFA-1在细胞表面保持均匀分布。相比之下,ICAM-1集中在未刺激的B淋巴母细胞在平面双层膜中黏附于LFA-1所通过的茎状结构中,但在添加佛波酯后几分钟内,ICAM-1免疫荧光在细胞表面变得更均匀分布。LFA-1和ICAM-1均未与褶皱膜区域中丝状肌动蛋白的显著染色共定位。通过细胞表面LFA-1和ICAM-1、2或3的相互作用促进了不同的细胞形态和行为,讨论了其与先前观察到的淋巴细胞与不同细胞类型相互作用模式的相关性。