Project 3: Interactions between microbial metabolites, airway pathogens and airway epithelial cell function

项目总结 早期生活环境改变了儿童过敏性疾病、喘息性疾病和哮喘的风险， 这些疾病在农场长大的儿童中不太常见。因此，在我们的研究中， 威斯康星州婴儿研究队列(WISC，由AADCRC计划资助)，特应性皮炎和 呼吸系统疾病与出生前和出生后早期农场暴露的强度有关 句号。值得注意的是，过敏和哮喘在传统农业(TA)环境中的儿童中很少见(例如，阿米什人 社区)，有强烈的农场暴露。早期生命中微生物的暴露和定居很可能有助于 通过促进上皮细胞功能和免疫发育而有益于健康。某些共生微生物 如棒状杆菌属。和Dolosigranulum pgrum与呼吸道细菌呈负相关 减少病原体，降低病毒性喘息性疾病和儿童哮喘恶化的风险。 这些发现表明，TA和健康农场儿童的呼吸道微生物群将是一个丰富的来源 可以抑制病毒和细菌病原体，提高上皮完整性，并提供抗病毒和抗病毒作用的微生物 病原体反应。为了验证这一假设，我们建立了体外培养的呼吸道上皮细胞模型。 细胞以确定呼吸道细菌、病毒感染和上皮细胞之间的相互作用机制 组成和功能。我们的初步数据表明，TA儿童具有微生物群落结构 富含具有不同遗传特征的“保护性”共生细菌。最后，我们使用了单细胞 转录组分鉴定高度易感的呼吸道上皮细胞的特征 鼻病毒(RV)，通常会在儿童时期引起喘息性疾病。 该项目将测试呼吸道共生细菌、呼吸道病原体和上皮细胞之间的相互作用。 细胞。首先，我们将从健康的TA和农场儿童中筛选共生菌，以寻找抑制病毒的代谢物 和细菌病原体。其次，我们将培养和分析来自TA、FARM和 非农儿童确定环境暴露是否会导致细胞内与群体相关的差异 抗菌肽的组成、抗病毒反应和合成。最后，我们将分析 从TA和农场儿童中选择细菌代谢物在培养的呼吸道上皮细胞上确定 它们在抗病原体反应中发挥有益的作用，并抑制T2炎症通路。 这些研究的结果可能导致新的治疗方法，带来一些健康益处 与农场对所有儿童的暴露有关。

PROJECT SUMMARY The early life environment modifies the risk for childhood allergic diseases, wheezing illnesses, and asthma, and these diseases are less common in children who grow up on farms. Accordingly, in our studies of the Wisconsin Infant Study Cohort ("WISC," funded by the AADCRC program), reductions in atopic dermatitis and respiratory illnesses were related to the intensity of farm exposures during the prenatal and early postnatal periods. Notably, allergies and asthma are rare in children in traditional agrarian (TA) settings (e.g., Amish communities) with intense farm exposures. Early life microbial exposures and colonization are likely to contribute to health benefits by promoting epithelial cell function and immune development. Certain commensal microbes such as Corynebacterium sp. and Dolosigranulum pigrum are inversely associated with bacterial respiratory pathogens and reduce the risks of viral wheezing illnesses and exacerbations of childhood asthma. These findings suggest that airway microbiomes of TA and healthy farm children will be a rich source of microbes that can inhibit viral and bacterial pathogens, increase epithelial integrity, and prime anti- pathogen responses. To test this hypothesis, we have established in vitro models of cultured airway epithelial cells to determine mechanisms for interactions between airway bacteria, viral infections, and epithelial cell composition and function. Our preliminary data indicate that TA children have microbiome community structures enriched for "protective" commensal bacteria with distinct genetic features. Finally, we used single-cell transcriptomics to identify characteristics of airway epithelial cells that are highly susceptible to infection with rhinoviruses (RV), which commonly cause wheezing illnesses during childhood. This project will test for interactions between airway commensal bacteria, airway pathogens, and epithelial cells. First, we will screen commensal bacteria from healthy TA and farm children for metabolites that inhibit viral and bacterial pathogens. Second, we will culture and analyze nasal airway epithelial cells from TA, farm, and non-farm children to determine whether environmental exposures lead to group-related differences in cell composition, antiviral responses, and synthesis of antimicrobial peptides. Finally, we will analyze effects of selected bacterial metabolites from TA and farm children on cultured airway epithelial cells to determine whether they exert beneficial effects on anti-pathogen responses and inhibit T2 inflammatory pathways. The results of these studies could lead to new therapeutic approaches to bring some of the health benefits associated with farm exposures to all children.