The human gut microbiome, malnutrition and vaccine responses

人类肠道微生物组、营养不良和疫苗反应

基本信息

批准号：
8469030
负责人：
Andrew Leon Kau
金额：
$ 6.2万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-04 至 2014-07-03
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8469030
关键词：
Address Adjuvant Age Agriculture Animals Antigens Archaea Area Bacteria Biological Assay Biological Markers Birth Cells Cessation of life Child Childhood Cholera Toxin Communities DNA Defect Development Diet Diet Monitoring Endotoxins Enteral Enzyme-Linked Immunosorbent Assay Excretory function Exhibits Failure Family Female Fetal Malnutrition Food Freezing Gene Expression Genes Germ-Free Glean Gnotobiotic Harvest Histology Human Immune Immune system Immunoglobulin A Immunoglobulin G Infant Infection Intervention Intestines Kwashiorkor Lactulose Lead Life Link Longitudinal Studies Macronutrients Nutrition Malawi Malnutrition Mannitol Messenger RNA Metabolic Metagenomics Methods Micronutrients Modeling Monitor Monozygotic Twinning Monozygotic twins Morbidity - disease rate Morphology Mothers Mucosal Immune Responses Mucosal Immunity Mus National Research Service Awards Neurodevelopmental Deficit Nutrient Nutritional Nutritional Support Nutritional status Oral Ovalbumin Partner in relationship Pathogenesis Pattern Physiological Pneumonia Population Predisposition Pregnancy Protocols documentation Recurrence Relative (related person)Ribosomal RNA Role Sampling Series Serum Shapes Shotguns Siblings Structure Surveys Testing Therapeutic Therapeutic Uses Time Translations Transplantation Twin Multiple Birth Vaccinated Vaccination Vaccine Design Vaccines Weaning Withdrawal absorption aged design disorder risk expectation feeding functional genomics global health gut microbiota immune function insight killings member microbial microbial community microbiome micronutrient deficiency mortality mouse model mucosal vaccination offspring postnatal pregnant pup pyrosequencing rRNA Genes research study response transcriptome sequencing transmission process

项目摘要

DESCRIPTION (provided by applicant): Malnutrition kills millions of children worldwide every year. Much of their morbidity and mortality is a result of infections, including those caused by enteropathogens. Efforts to stop these infections have been limited in some populations by unexplained vaccine failures. I hypothesize that there is an interrelationship between malnutrition, the gut microbiome, and the immune system that is causally linked with vaccine responsiveness and that these interrelationships can be informatively probed using 'humanized' gnotobiotic mice. Our lab is conducting longitudinal studies of the gut microbiomes of twins aged 1-36 months living in Malawi who are healthy, or who are concordant or discordant for severe forms of malnutrition (e.g., kwashiorkor). We are addressing the following questions: Are there identifiable configurations of the gut microbiome associated with severe forms of malnutrition? How is the microbiota/microbiome reconfigured with a Ready to Use Therapeutic Food (RUTF) intervention and does it persist after cessation of RUTF? If the configured state does not persist after withdrawal of RUTF, does this mean that in the absence of longer-term nutritional support, lingering abnormalities in the metabolic activities contribute to persistent physiological abnormalities in the human host? We have recently shown that it is possible to efficiently transplant the microbial community present in frozen human fecal samples into germ-free (GF) mice and to transmit the engrafted microbiota from mothers to their offspring. With this capability, I propose two aims for this NRSA application. Aim 1. Use gnotobiotic mice, colonized at 6 weeks of age with de-identified fecal microbiota already collected from several sets of monozygotic twins discordant for kwashiorkor (1 healthy/ 1 malnourished) as well as their mothers' microbiota. Recipients will be fed a macro- and micronutrient deficient Malawi diet, or a control nutritionally 'sufficient' diet. I will follow their responses to mucosal vaccination with cholera toxin and ovalbumin (a model mucosal adjuvant/antigen) using antigen-specific ELISA. I will define their gut mucosal barrier function by serum LPS assays, lactulose/mannitol absorption assays, histo- and immunohistochemical assays of gut morphology and immune cell populations (exploring whether they have features of tropical enteropathy), plus functional genomic studies of gut gene expression. I will perform time series metagenomic studies on their fecal microbiomes (sequencing bacterial 16S rRNA genes, total community DNA, and expressed mRNAs). Aim 2. Assess the role of maternal nutritional and microbiota 'status' on the vaccine responses of their offspring by colonizing pregnant female GF mice with fecal samples from twins discordant for kwashiorkor and their mothers. Mothers will be maintained on either a Malawi or nutrient- sufficient control diet prior to, during and after pregnancy; their offspring will be weaned onto nutrient- deficient or -sufficient diets and subjected to the same vaccination protocols and analyses as described in Aim 1. These experiments should help identify biomarkers of malnutrition and vaccine responsiveness.

描述（由申请人提供）：营养不良每年杀死全球数百万儿童。他们的大部分发病率和死亡率是感染的结果，包括由肠病毒引起的感染。在某些人群中，无法解释的疫苗失败的努力受到限制。我假设营养不良，肠道微生物组和与疫苗反应性有因果关系的免疫系统之间存在相互关系，并且可以使用“人源化” gnotobiotic小鼠对这些相互关系进行有因果关系。我们的实验室正在对居住在马拉维健康的1-36个月的双胞胎的肠道微生物组进行纵向研究，他们健康，或者对严重的营养不良形式一致或不一致（例如，kwashiorkor）。我们正在解决以下问题：是否存在与严重营养不良形式相关的肠道微生物组的可识别构型？微生物群/微生物组如何通过准备使用治疗食品（RUTF）干预重新配置，并且在RUTF停止后是否持续存在？如果配置状态在RUTF退出后不持续，这是否意味着在没有长期营养支持的情况下，代谢活动中的异常异常有助于人类宿主的持续生理异常？我们最近表明，有可能有效地将存在的人类粪便样品中存在的微生物群落移植到无菌（GF）小鼠中，并将植入的微生物群从母亲传播到其后代。有了这个功能，我提出了针对此NRSA应用的两个目标。 AIM 1。使用gnotobiotic小鼠，在6周龄殖民时，已经从几组单组的单卵双胞胎双胞胎（1个健康/ 1个营养不良）以及母亲的Microbobiota中收集了几组单卵双胞胎。接收者将获得宏观和微量营养素不足的马拉维饮食，或者在营养上“足够”的饮食中喂养。我将遵循他们使用抗原特异性ELISA的霍乱毒素和卵蛋白（一种模型粘膜辅助/抗原）对粘膜疫苗接种的反应。我将通过血清LPS分析，乳乳糖/甘露糖吸收测定法，组织和免疫组织化学测定法（探索它们是否具有热带肠病特征）以及Gut Gene表达的功能性基因组研究，来定义其肠粘膜屏障功能。我将对其粪便微生物组（测序细菌16S rRNA基因，总群落DNA和表达的mRNA）进行时间序列的核类研究。 AIM 2。通过将怀孕的雌性GF小鼠与来自Kwashiorkor及其母亲的双胞胎的粪便样本一起定居于怀孕的雌性GF小鼠，评估母体营养和微生物群“状态”对后代的疫苗反应的作用。在怀孕之前和之后，母亲将维持在马拉维或营养足够的控制饮食上；它们的后代将被断奶为营养不足或富含饮食，并接受与AIM 1所述的相同疫苗接种方案和分析。这些实验应有助于识别营养不良和疫苗反应性的生物标志物。