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MECHANISMS OF BACTERIAL ADHERENCE & COLONIZATION

细菌粘附机制

基本信息

批准号：
2129670
负责人：
DAVID L HASTY
金额：
$ 12.41万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-04-01 至 1995-11-30
项目状态：
已结题

项目摘要

The long-term goal of this project is to understand the role of host macromolecules in bacterial colonization of the mucosal surfaces of the oropharynx. We will focus principally on the role salivary molecules play in the modulation of adherence of type 1 fimbriated gram negative bacteria frequent agents of nosocomial pneumonia) to mucosal epithelial cells. We will purify type 1 fimbrial-binding glycoproteins (FBGs) from saliva by routine chromatographic procedures. We will also test sample of known salivary glycoproteins obtained from other investigators in the field. The FBGs will be characterized by amino acid analysis, two- dimensional SDS polyacrylamide gel electrophoresis, peptide mapping, and/or immunological crossreactivity. Oligosaccharides will be characterized by chemical and lectin-binding assays and, where warranted, by mass spectrometry and nuclear magnetic resonance spectrometry. The specific antibody probes generated will be used to assay for the concentration of the various molecules in saliva. Purified FBGs will be radiolabeled and used to determine the specificity, affinity and number of binding sites on type 1 fimbriated E. coli and FN. Appropriate controls will include E. coli mutants which lack or overproduce the 29 kDa mannose-binding adhesin and other species which express type 1 adhesins. Attention will be given to the influence of pH, ionic strength and divalent cations. Competitive inhibition studies using FN will be used to further characterize these interactions. Radiolabeled FBGs will also be used in binding studies to determine the specificity, affinity and number of binding sites on epithelial cells. Attention will be given to the influence of pH, ionic strength, divalent cations and to the effects of saliva and FN on this interaction. Since the interaction of FBGs with type 1 fimbriae may be mediated through the oligosaccharide portion of the glycoprotein, we will study the effects of glycosidases on the interaction of E. coli with the purified FBGs and determine whether exposure of saliva to these glycosidases exposes additional FBGs. We will use an animal model to study the hypothesis that the increased gram negative colonization following trauma is due, at least in part, to changes in salivary or buccal cell-associated molecules. Samples of epithelial cells and saliva will be assayed for: 1) changes in E. coli (or other gram negative bacteria) adherence using standard adherence assays, 2) changes in levels of FBGs, Fn or Fn fragments on buccal cells and in saliva using immuno-fluorescence, Western blot analysis and quantitative immunological assays, and 3) changes in the levels of FBG- or Fn- degrading activity using will add significantly to the information needed to understand important aspects of bacterial pathogenesis in the oropharyngeal cavity.

这个项目的长期目标是理解的角色

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Assay for adhesion of host cells to immobilized bacteria.

宿主细胞与固定化细菌粘附的测定。

DOI：
10.1016/s0076-6879(95)53021-5
发表时间：
1995
期刊：
Methods in enzymology
影响因子：
0
作者：
Sokurenko,EV;Hasty,DL
通讯作者：
Hasty,DL

Characterization of unique human TCR V beta specificities for a family of streptococcal superantigens represented by rheumatogenic serotypes of M protein.

以风湿性 M 蛋白血清型为代表的链球菌超抗原家族的独特人类 TCR V beta 特异性的表征。

DOI：
发表时间：
1994
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
0
作者：
Watanabe-Ohnishi,R;Aelion,J;LeGros,L;Tomai,MA;Sokurenko,EV;Newton,D;Takahara,J;Irino,S;Rashed,S;Kotb,M
通讯作者：
Kotb,M

Pathogenesis of experimental endocarditis.

实验性心内膜炎的发病机制。