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Study on the "Streptococcus milleri" Adhesins Involved in Its Colonization to Human Bodies

“米勒链球菌”粘附素及其在人体定植的研究

基本信息

批准号：
08457575
负责人：
INOUE Masakazu
金额：
$ 4.61万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

Of the 36 "Streptpcoccus milleri" strains tested of various serotypes, only 26 serotype g, g-related or h Streptpcoccus intermedius, Streptococcus constellatus or Streptococcus constellatus strains (21,4 and 1 strains respectively) produced cellular agglutination in human whole saliva filtrate. Cells of S.intermedius 1208-1 (serotype g) but not of S.constellatus NCTC 10708 (b) carried long fibrils which extended radially from cell surface and assumed a curly single strand of 1 mum or longer, suggesting that saliva-indeced cell-aggregating factor (s) may exist on the peritrichous appendage. The fibrils were isolated from cells by sonication and purified by 50% ammonium sulfate precipitation followed by a DEAE-Sepharose CL-6B column chromatography. The purified fibrils were composed of carbohydrate and protein in an approximate weight ratio of 3 : 1. Serine, cysteine and lysine were contained in relatively high proportions (10.0-7.9 mol%) and nonpolar amino acids were detected 38.1 mol%. Carbohydrate moiety was composed of mannose, galactose and their amino derivatives at an approximate molar ratio of 4 : 2 : 1. The fibrillar materials were partially hydrolyzed by papain, pronase-P and subtilisin but not by any glycosidases tested. The purified fibrils possessed the molecular weightof higher than 250 kd and were suggested to be composed of unit monomer of 60kd. The cellular aggregation was inhibited by heat treatment of cells and by the presence of the anti-fibril immunoglobulin Fab fragment or the purified fibril preparation. However, the heat-sensitive cellular aggregation-mediating site was supposed not to be identical to the heat-resistant antigenic determinant of fiblillar molecules. Of the 26 saliva-induced aggregating strains (g, h or g-related), only 20 S.intermedius, S.costellatus or S.anginosus strains, but 6 S.intermedius or S.constellatus strains did not, produced the particular type of fibril detected with S.intermedius 1208-1 (g) cells.

在36株不同血清型的“米勒链球菌”中，只有26株（分别为21株、4株和1株）血清型G、G相关或h中间链球菌、星座链球菌或星座链球菌菌株在人全唾液滤液中产生细胞凝集。中间链球菌1208-1（血清型g）的细胞携带长纤维，从细胞表面放射状延伸，呈1 μ m或更长的卷曲单链，而星座链球菌NCTC 10708（血清型B）的细胞则不携带长纤维，这表明唾液诱导的细胞聚集因子可能存在于周毛附件上。通过超声处理从细胞中分离原纤维，并通过50%硫酸铵沉淀，然后通过DEAE-Sepharose CL-6 B柱层析纯化。纯化的原纤维由碳水化合物和蛋白质以大约3：1的重量比组成。丝氨酸、半胱氨酸和赖氨酸以相对高的比例（10.0-7.9摩尔%）包含，并且检测到非极性氨基酸38.1摩尔%。碳水化合物部分由甘露糖、半乳糖及其氨基衍生物组成，摩尔比约为4：2：1。纤维状材料被木瓜蛋白酶、链霉蛋白酶-P和枯草杆菌蛋白酶部分水解，但不被任何测试的糖苷酶水解。纯化后的纤维分子量大于250 kd，由60 kd的单体组成。通过细胞的热处理和通过抗原纤维免疫球蛋白Fab片段或纯化的原纤维制备物的存在抑制细胞聚集。然而，热敏感的细胞聚集介导的网站被认为是不相同的耐热抗原决定簇的纤维分子。在26种唾液诱导的聚集菌株（g、h或g相关）中，只有20种中间链球菌、中星链霉菌或咽峡炎链霉菌菌株产生了用中间链球菌1208-1（g）细胞检测到的特定类型的原纤维，但6种中间链球菌或星座链霉菌菌株没有产生。