Characterizing the Contribution of the Group B Streptococcal Surface Adhesin BspC Interaction with Host Vimentin to Disease and Colonization

表征 B 族链球菌表面粘附素 BspC 与宿主波形蛋白相互作用对疾病和定植的贡献

• 批准号: 10312484
• 负责人: Haider Syed Manzer
• 金额: $ 3.4万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10312484
• 关键词: Address; Adherence; Adult; Alanine; Amino Acids; Antibiotic Therapy; Bacteria; Bacterial Adhesins; Bacterial Infections; Bacterial Meningitis; Binding; Binding Sites; Biological Assay; Birth; Blood; Blood - brain barrier anatomy; Blood Vessels; Brain; CXCL1 gene; CXCL2 gene; Cell Communication; Cell Line; Cells; Central Nervous System Diseases; Cerebral Ischemia; Cervical; Cervix Uteri; Characteristics; Chromosomes; Cognitive deficits; Complement; Data; Development; Disease; Disease Progression; Endometritis; Endothelial Cells; Endothelium; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Etiology; Exhibits; Family; Female; Female genitalia; Fetus; Gastrointestinal tract structure; Human; Hydrophobicity; I-antigen; IL6 gene; IL8 gene; Immune response; In Vitro; Infant; Infection; Inflammation; Inflammatory; Interleukin-1 beta; Intermediate Filament Proteins; Invaded; Knockout Mice; Length; Ligand Binding; Measures; Meningeal; Meningitis; Molecular; Mus; Mutation; Neonatal meningitis; Neuraxis; Neurologic; Newborn Infant; Pathogenesis; Plasmids; Point Mutation; Population; Pregnancy; Pregnant Women; Premature Birth; Premature Labor; Proteins; Publications; Quantitative Reverse Transcriptase PCR; Recombinants; Role; Seizures; Signal Transduction; Site-Directed Mutagenesis; Streptococcus Group B; Surface; Survivors; System; Tertiary Protein Structure; Testing; Therapeutic; Tissues; Transcript; Uterus; Vagina; Vimentin; Woman; blood-brain barrier penetration; brain endothelial cell; cell type; design; epithelial to mesenchymal transition; experimental study; in silico; in utero; in vivo; in vivo Model; insight; intraamniotic infection; intrauterine infection; mortality; mouse model; mutant; neonatal bacterial meningitis; neonatal infection; neonate; pathogenic bacteria; placental membrane; prevent; reproductive tract; screening; transmission process; treatment strategy; urogenital tract; vaccine access; vector; yeast two hybrid system

PROJECT SUMMARY Group B Streptococcus (GBS, also known as Streptococcus agalactiae) is a Gram-positive, -hemolytic bacterium normally found in the human gastrointestinal and urogenital tracts. GBS remains a leading cause of invasive disease in newborns and certain adult populations including pregnant women. The development of GBS disease is initiated by the asymptomatic colonization of the female genital tract and during pregnancy can be associated with chorioamnionitis, puerperal endometritis and preterm labor. Approximately 20-30% of healthy women are colonized rectovaginally with GBS; the majority of infants born to these women will themselves become colonized with the bacterium. Newborn infection also results from ascending infection of the bacterium through the placental membranes to initiate infection in utero. GBS is the principle etiologic agent of neonatal bacterial meningitis, and GBS meningeal infection results in 10-15% mortality despite antibiotic treatment. Additionally, up to 40% of survivors develop permanent neurological sequelae, including cognitive defects, seizure activity, and cerebral ischemia. To access the central nervous system (CNS) and cause meningitis blood-borne GBS must penetrate the blood-brain barrier (BBB); however, little is known about the very first and crucial interaction between GBS and the BBB that initiates bacterial crossing and disease progression. This proposal seeks to characterize the role of a newly identified GBS Antigen I/II family adhesin, BspC, to the pathogenesis of colonization and CNS disease. We have recently determined that BspC promotes attachment to BBB endothelium and contributes to the development of GBS meningitis. Further we have discovered that BspC interacts directly with vimentin, a widely distributed intermediate filament protein found in blood vessel endothelial cells. Thisproposal seeks toidentify the region on BspC that interacts with vimentin, and to characterize the role of this interaction in GBS meningitis and colonization of the female reproductive tract. I hypothesize that the binding region is contained within the globular variable domain (V- domain) of BspC and contributes to both GBS meningitis and vaginal colonization/ascending infection. These hypotheses will be addressed with both in vitro and in vivo models of BBB penetration and vaginal colonization in the following specific aims: Aim 1: Identify the critical regions and amino acids required for BspC V-domain- vimentin interaction; Aim 2: Characterize the role of BspC and vimentin in the female reproductive tract. These data will clarify the crucial role of Antigen I/II proteins to GBS colonization and disease and provide fundamental mechanistic insights that may inform new treatment strategies to prevent colonization and ultimately bacterial meningitis.

项目摘要 B族链球菌（GBS，也称为无乳链球菌）是革兰氏阳性、溶血性链球菌， 通常在人类胃肠道和泌尿生殖道中发现的细菌。GBS仍然是导致 新生儿和某些成年人群（包括孕妇）的侵袭性疾病。的发展 GBS疾病是由女性生殖道的无症状定植引起的，在怀孕期间， 与绒毛膜炎、产褥期子宫内膜炎和早产有关。约20-30% 健康妇女的直肠阴道定植有GBS;这些妇女所生的大多数婴儿将 它们自身也被细菌定殖。新生儿感染也是由 细菌通过胎盘膜在子宫内引发感染。GBS是主要病因 新生儿细菌性脑膜炎的病原体，GBS脑膜感染导致10-15%的死亡率， 抗生素治疗此外，高达40%的幸存者发展为永久性神经系统后遗症，包括 认知缺陷癫痫发作和脑缺血进入中枢神经系统（CNS）， 引起脑膜炎的血源性GBS必须穿透血脑屏障（BBB）;然而，对血脑屏障的作用知之甚少。 GBS和BBB之间的第一个和关键的相互作用，引发细菌交叉和疾病 进展该提议试图描述新鉴定的GBS抗原I/II家族粘附素的作用， BspC与定植和CNS疾病的发病机制有关。我们最近确定BspC 促进对BBB内皮的附着并有助于GBS脑膜炎的发展。我们越 我发现BspC直接与波形蛋白相互作用，波形蛋白是一种广泛分布的中间丝蛋白 发现于血管内皮细胞。该提案旨在确定BspC上与以下方面相互作用的区域： 波形蛋白，并描述这种相互作用在GBS脑膜炎和女性定植中的作用。 生殖道我假设结合区包含在球状可变结构域（V-）内。 结构域），并有助于GBS脑膜炎和阴道定植/上行感染。这些 假设将通过血脑屏障渗透和阴道定植的体外和体内模型来解决 目的1：鉴定BspC V-结构域所需的关键区域和氨基酸， 波形蛋白相互作用;目的2：表征BspC和波形蛋白在女性生殖道中的作用。这些 这些数据将阐明抗原I/II蛋白对GBS定植和疾病的关键作用， 基本的机制见解，可以为新的治疗策略提供信息，以防止殖民化， 最终导致细菌性脑膜炎。