Dehio C, Freissler E, Lanz C, Gómez-Duarte O G, David G, Meyer T F
Max-Planck-Institut für Biologie, Abteilung Infektionsbiologie, Spemannstrasse 34, Tübingen, D-72076, Germany.
Exp Cell Res. 1998 Aug 1;242(2):528-39. doi: 10.1006/excr.1998.4116.
Binding of a particular opacity outer membrane protein (Opa) of Neisseria gonorrhoeae to cell surface heparan sulfate proteoglycans (HSPGs) of epithelial cells results in tight bacterial adherence; however, the role of this ligand-receptor interaction in triggering the subsequent bacterial internalization step is uncertain. Here we have used latex beads coated with HSPG-ligating antibodies as an in vitro model to study the role of HSPGs in gonococcal uptake into epithelial cells. Beads and gonococci showed the same cell line-specified adherence patterns and increase in phagocytic uptake mediated by serum or purified vitronectin (Vn). Heparitinase digestion as well as antibody competition experiments indicate that a critical level of HSPG ligation is necessary and sufficient to trigger phagocytic uptake into epithelial cells. Vn was found to specifically enhance HSPG-dependent phagocytic uptake while phagocytosis resulting from the ligation of other cell surface receptors was unaffected in the presence of Vn. Pharmacological studies with PKC inhibitors suggest a role for PKC in phagocytic uptake of HSPG-ligating beads. The use of drugs impairing cytoskeletal functions indicates that HSPG-dependent phagocytosis requires actin polymerization by a process distinct from receptor-mediated endocytosis.
淋病奈瑟菌特定的不透明外膜蛋白(Opa)与上皮细胞表面硫酸乙酰肝素蛋白聚糖(HSPG)结合会导致细菌紧密黏附;然而,这种配体-受体相互作用在触发随后细菌内化步骤中的作用尚不确定。在此,我们使用包被有HSPG连接抗体的乳胶珠作为体外模型,来研究HSPG在淋球菌摄取到上皮细胞中的作用。珠子和淋球菌表现出相同的细胞系特异性黏附模式,并且由血清或纯化的玻连蛋白(Vn)介导的吞噬摄取增加。硫酸乙酰肝素酶消化以及抗体竞争实验表明,HSPG连接达到临界水平对于触发上皮细胞的吞噬摄取是必要且充分的。发现Vn可特异性增强HSPG依赖的吞噬摄取,而在存在Vn的情况下,由其他细胞表面受体连接导致的吞噬作用不受影响。使用蛋白激酶C(PKC)抑制剂进行的药理学研究表明,PKC在HSPG连接珠的吞噬摄取中起作用。使用损害细胞骨架功能的药物表明,HSPG依赖的吞噬作用需要通过与受体介导的内吞作用不同的过程进行肌动蛋白聚合。