Microecology-murine gut-initiation & progression of IBD

描述(申请人提供)：具有免疫功能的转基因小鼠 在试图解体方面，监管失调的做法越来越成功 并理解人类中的IBD。IBD被认为是一种 遗传和环境因素的结合。微生物区系有 无疑是疾病过程的一个重要组成部分，在这个过程中，微生物 抗原被认为引发和促进炎症，特别是在 存在免疫调节失调或粘膜屏障受损 易感宿主。对远端复杂微生态的认识 肠道是极其有限的，因为辨认的繁琐性质 通过常规方法检测单个细菌种类和菌株，以及 不能培养许多挑剔的共生生物。为了绕过这一点 缺乏细菌物种，已知定植的灵知菌动物 微生物区系已被利用为极大的优势。一种标准化的微生物区系，用于 将无菌啮齿动物定殖化称为“改变的舍德勒菌群”(ASF)。 发展起来的。由于体内监测系统的局限性， 鉴定ASF中的8种厌氧细菌，我们最近表征了 利用16S rRNA确定它们相对于已知细菌的系统发育位置 序列分析。这项提案将使用分子技术，基于 ASF 16S rRNA的定量聚合酶链式反应，以筛选微生物多样性 小鼠肠道和确定微生物区系动力学，根据定义的实验 条件，无论有没有螺杆菌，都会影响启动和 炎症性肠病在小鼠体内的进展。具体地说，我们将1)使用 分子技术，鉴定，定量和分配 IL-10-/-胃肠道改变的舍德勒菌群(ASF) 还有C57BL小鼠。2)确定肠道致病性幽门螺杆菌是如何。 LL-10-/-小鼠伴发的结肠炎改变了小鼠的微生态。 下肠道，相应地，确定单个物种是否以及如何 血管紧张素转换酶对慢性肠道疾病进展或消退的影响 发炎。3)测定ASF在生理性和非生理性条件下的微生物动态 小鼠下肠道的解剖紊乱及宿主改变 这些变化如何影响随后的伤寒结肠炎的诱发 可能是由幽门螺杆菌引起的结肠腺癌。4)确定 ASF和幽门螺杆菌的扰动。在口头之前和之后 接种幽门螺杆菌疫苗。抗原和粘膜佐剂，阐明 这些疫苗策略如何影响微生物区系动态和影响肠道 细胞因子反应。

DESCRIPTION (provided by applicant): Genetically engineered mice with immune dysregulation have been used with increasing success in attempting to unravel and comprehend IBD in humans. IBD is considered to be the result of a combination of genetic and environmental factors. Microbial flora are undoubtedly an important component of the disease process, in which microbial antigens are thought to initiate and promote inflammation, particularly in the presence of immune dysregulation or an impaired mucosal barrier in the susceptible host. The understanding of the complex microecology of the distal intestine is extremely limited because of the tedious nature of identifying individual bacterial species and strains by conventional methods, and the inability to culture many fastidious commensal organisms. To circumvent this lack of bacterial speciation, gnotobiotic animals colonized with known microbiota have been used to great advantage. A standardized microbiota used in colonizing germfree rodents referred to "Altered Schaedler Flora" (ASF) was developed. Because of the limitations of an in vivo monitoring system used to identify the 8 anaerobic bacterial species in ASF, we recently characterized their phylogenetic positions relative to known bacteria by utilizing 16S rRNA sequence analysis. This proposal will use molecular techniques, based on quantitative PCR of ASF 1 6S rRNA, to screen the microbial diversity of the murine gut and ascertain how microflora dynamics, under defined experimental conditions, with and without helicobacters, influence initiation and progression of IBD in the mouse. Specifically we will 1) characterize, using molecular techniques, the identification, quantification and distribution of Altered Schaedler's Flora (ASF) in the gastrointestinal tract of the IL-1O-/- and the C57BL mouse. 2) Determine how enteropathogenic Helicobacter spp. associated typhlocolitis in the lL-1O-/- mice alters the microecology of the lower bowel and correspondingly, ascertain whether and how individual species in ASF influence the progression or attenuation of chronic intestinal inflammation. 3) Determine microbial dynamics of ASF after physiological and anatomical perturbation of the lower bowel in mice as well as altering host genotype an how these changes influence subsequent induction of typhlocolitis and possibly colonic adenocarcinoma induced by Helicobacter spp. 4) Determine perturbations of ASF and Helicobacter spp. prior and subsequent to oral vaccination with Helicobacter spp. antigens and mucosal adjuvants, elucidate how these vaccine strategies influence microflora dynamics and impact gut cytokine responses.