Bacterial colonization and immunogenicity in the mosquito gut ecosystem

描述（由申请人提供）：肠道微生物组对于引发蚊子基础免疫至关重要。作为了解蚊子肠道生态系统中微生物组的一项努力，该项目解决了有关非致病性细菌如何在肠道中定植并与宿主免疫监视相互作用以维持共生关系的重要问题，从而有效保护免受疟疾等病原体的侵害。细菌细胞壁 组分脂多糖（LPS）使定植和免疫原性联系起来。我们已经分离出一种主要的肠道细菌肠杆菌属，从蚊子的肠道，并产生突变体，其中基因waaL被破坏。突变株的LPS存在缺陷。我们将使用野生型和waal突变体来研究LPS在肠道细菌定植中的作用及其在免疫原性效力中的作用。我们将使用GFP或RFP标记的肠杆菌野生型或突变体建立体内工作模型，以重建肠道种群，然后确定其定殖效率。RNA-seq将用于同时分析肠杆菌和肠道的转录组，这是一种双重方法，用于获取与细菌和宿主之间相互作用相关的数据。还将评估LPS缺陷对细菌免疫原性和宿主抗疟疾免疫力的影响。该项目将产生数据，以促进我们了解共生微生物群落是如何形成的，以及宿主免疫系统如何与共同进化的非致病微生物相互作用，以维持肠道生态系统中的共生。这些知识将有助于开发新的方法来加强蚊子抗疟疾免疫力，以减少疟疾传播。

DESCRIPTION (provided by applicant): Mosquito gut microbiome is critical for priming mosquito basal immunity. As an effort to understand microbiome in the mosquito gut ecosystem, this project addresses important questions regarding how non-pathogenic bacteria colonize the gut and interact with host immune surveillance to maintain a symbiotic relationship, which in turn provides effective protection against pathogens like malaria. The bacterial cell wall component lipopolysaccharide (LPS) makes a connection of colonization and immunogenicity. We have isolated a predominant gut bacterium Enterobacter sp., from the mosquito gut, and a mutant was generated, in which the gene waaL was disrupted. The LPS of the mutant strain has defects. We will use the wild type and the waal mutant to study the role of LPS in bacterial colonization of the gut and its role in immunogenic potency. We will create an in vivo working model using GFP- or RFP- tagged Enterobacter wild type or mutant to reconstruct the gut population, and then determine its efficiency of colonization. The RNA-seq will be used for analyzing transcriptomes of both Enterobacter and the gut simultaneously, a dual approach to acquire data pertinent to the mutual interactions between the bacteria and host. The impact of the LPS defects on bacterial immunogenicity and host immunity against malaria will also be assessed. The project will generate data to advance our understanding how a symbiotic microbial community is shaped, and how host immune system interacts with co-evolved non-pathogenic microorganisms to maintain symbiostasis in the gut ecosystem. The knowledge will facilitate to develop novel ways to strengthen mosquito anti-malaria immunity to reduce malaria transmission.