Valentin-Weigand P, Grulich-Henn J, Chhatwal G S, Müller-Berghaus G, Blobel H, Preissner K T
Institute for Bacteriology and Immunology, Max-Planck-Gesellschaft, Justus-Liebig-Universität, Giessen, Federal Republic of Germany.
Infect Immun. 1988 Nov;56(11):2851-5. doi: 10.1128/iai.56.11.2851-2855.1988.
The role of S protein in the adherence of group A and G streptococci to human umbilical vein endothelial cells cultivated in 96-well microdilution plates was studied by utilizing fluorescein-labeled streptococci. The assay proved suitable for quantitative determination of bacterial adherence to cultured endothelial cells for all tested strains of streptococci. Only bacterial strains with significant S protein binding but weak fibronectin binding were included in these studies. Fibronectin-mediated adherence to endothelial cells of these streptococci was less than 25% of total and could be blocked by antifibronectin immunoglobulin G. Further treatment of endothelial cell monolayers with anti-S protein immunoglobulin G at concentrations up to 1 mg per well led to an almost complete inhibition of adherence for all tested streptococcal cultures, indicating significant contribution of S protein in the streptococcus-endothelial cell interaction. Blocking of S-protein-binding sites on streptococci by preincubation with exogeneous S protein at a concentration of 10 micrograms per 4 x 10(7) streptococci led to about 75% reduction of S-protein-mediated adherence to endothelial cells. Trypsin pretreatment of group G streptococci and pronase pretreatment of group A and G streptococci, modifications known to destroy the bacterial binding sites of S protein, also inhibited the capacity of the streptococci for S-protein-mediated adherence to endothelial cells by 75 to 80%. These results indicate that S protein plays a mediatory role in adherence of streptococci to endothelial cells and that S-protein-specific binding sites on streptococci are involved in this interaction.
利用荧光素标记的链球菌,研究了A组和G组链球菌的S蛋白在96孔微量稀释板中培养的人脐静脉内皮细胞黏附过程中的作用。该检测方法被证明适用于定量测定所有测试链球菌菌株对培养内皮细胞的细菌黏附。这些研究仅纳入了具有显著S蛋白结合但纤连蛋白结合较弱的细菌菌株。这些链球菌通过纤连蛋白介导的对内皮细胞的黏附占总量的比例不到25%,并且可被抗纤连蛋白免疫球蛋白G阻断。用浓度高达每孔1毫克的抗S蛋白免疫球蛋白G进一步处理内皮细胞单层,导致所有测试链球菌培养物的黏附几乎完全受到抑制,这表明S蛋白在链球菌与内皮细胞的相互作用中起重要作用。用浓度为每4×10⁷个链球菌10微克的外源S蛋白预孵育,阻断链球菌上的S蛋白结合位点,导致S蛋白介导的对内皮细胞的黏附减少约75%。对G组链球菌进行胰蛋白酶预处理以及对A组和G组链球菌进行链霉蛋白酶预处理(已知这些修饰会破坏S蛋白的细菌结合位点),也使链球菌通过S蛋白介导黏附于内皮细胞的能力降低了75%至80%。这些结果表明,S蛋白在链球菌对内皮细胞的黏附中起介导作用,并且链球菌上的S蛋白特异性结合位点参与了这种相互作用。