Safety and Effect of L. reuteri on Biomarkers of Inflammation in Healthy Infants

罗伊氏乳杆菌的安全性及其对健康婴儿炎症生物标志物的影响

基本信息

批准号：
8304030
负责人：
JON Marc RHOADS
金额：
$ 11.4万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8304030
关键词：
Abdominal colic Adult Affect B-Lymphocytes Bacteria Basic Science Biological Markers Blood Cattle Cells Characteristics Clinical Clinical Markers Crying Data Dose Double-Blind Method Drops Effector Cell Enrollment Escherichia coli Evolution Feces Flow Cytometry Future Hand Health Hour Incidence Infant Inflammation Inflammatory disease of the intestine Intestines Investigation Klebsiella Laboratories Lactobacillus Lactobacillus reuteri Leukocyte L1 Antigen Complex Leukocytes Life Lipopolysaccharides Measurement Measures Mediating Milk Modeling Molecular Mononuclear Necrotizing Enterocolitis Neonatal Newborn Infant Oral cavity Organism Outcome Peripheral Blood Mononuclear Cell Phase Placebo Control Play Population Postpartum Depression Prevalence Probiotics Production Publishing Randomized Controlled Trials Rattus Regulatory T-Lymphocyte Role Safety Serum Severities Shaken baby syndrome Stress T-Lymphocyte T-Lymphocyte Subsets Techniques Time Toll-like receptors automobile accident cytokine feeding gastrointestinal improved mortality novel pup receptor expression volunteer

项目摘要

DESCRIPTION (provided by applicant): Colic is a severe problem characterized by fussing, crying and screaming for more than 3 hours daily and affecting ~15% of otherwise normal infants in the first 3 months of life. We recently published our trial defining key clinical and microbiological characteristics of infants with colic. Compared to normal infants, those with colic demonstrated a four-fold increase in crying time, a doubling in the level of fecal calprotectin (a measure of gut inflammation), and changes in the fecal microbiota characterized by reduced bacterial diversity and an increase in the prevalence of Klebsiella. These results suggested a novel hypothesis: that the infants are crying because their bowels are inflamed, possibly as a result from an abnormal microbiota. Our investigation will focus on a population of healthy infants with colic to show safety of a probiotic that has been shown to reduce crying time in infants with colic. Our basic science laboratory trials showed that feeding of L. reuteri reduced gut inflammation and reduced mortality by > 50% in a neonatal rat model of necrotizing enterocolitis. We are currently close to completion of a double-blind placebo-controlled Phase 1 safety trial of L. reuteri in adult volunteers (U01AT003519). In this study, we are measuring how the probiotic influences the effector cells that modulate inflammation. We are quantifying circulating T cells, B cells, mononuclear cells, T cell subsets (by flow cytometry). We are measuring cytokine production by peripheral blood mononuclear cells, fecal calprotectin, and fecal microbiota. This trial includes measurement of regulatory T-cells (Tregs), the cells proposed to mediate the immunomodulatory effects of probiotics. Thus, our Primary Aim is to demonstrate (in a two-year trial) if Lactobacillus reuteri is safe in healthy newborns with colic (n=45), at 3 different doses: 5 x 106, 5 x 107, and 5 x 108 c.f.u. given by mouth once daily for si weeks. Our Secondary Aim is to compare outcomes to determine the optimal dose for a future randomized controlled trial (RCT). Dose-effect relationships will be determined for the following measures: (a) crying + fussing time; (b) fecal calprotectin; (c) serum cytokines; (d) toll-like receptor expression on peripheral blood mononuclear cells; (e) circulating regulatory T-cells; and (f) fecal levels of colic-related organisms (Klebsiella and E. coli, determined by qPCR) and diversity of fecal microbiota (assessed by 454 pyrosequencing). PUBLIC HEALTH RELEVANCE: Colic is a severe condition affecting ~ 15% of otherwise normal infants and is associated with postpartum depression, marital stress, and shaken baby syndrome. Babies that we studied in Houston cried for an average of 5 hours daily and had intestinal inflammation associated with an abnormal population of resident bacteria (microbiota) in their stools. Our study will determine the safety of a probiotic (health-promoting bacteria) called Lactobacillus reuteri in normal babies with colic. Furthermore, we will determine if Lactobacillus reuteri influences crying/fussing time, alters measurements of gut inflammation (in the stool and blood), and/or changes the resident fecal microbiota (using molecular techniques).

描述（由申请人提供）：绞痛是一种严重的问题，其特征是每天烦躁、哭泣和尖叫超过 3 小时，影响约 15% 的正常婴儿在出生后的前 3 个月内。我们最近发表了我们的试验，定义了绞痛婴儿的关键临床和微生物学特征。与正常婴儿相比，患有肠绞痛的婴儿结果表明，哭泣时间增加了四倍，粪便钙卫蛋白（肠道炎症的一种衡量指标）水平加倍，粪便微生物群发生了变化，其特征是细菌多样性减少和克雷伯氏菌患病率增加。这些结果提出了一个新的假设：婴儿哭泣是因为他们的肠道发炎，可能是由于微生物群异常所致。我们的研究将重点关注患有绞痛的健康婴儿群体，以证明益生菌的安全性，该益生菌已被证明可以减少绞痛婴儿的哭泣时间。我们的基础科学实验室试验表明，在坏死性小肠结肠炎新生大鼠模型中，喂养罗伊氏乳杆菌可减少肠道炎症并将死亡率降低 50% 以上。我们目前即将完成对成年志愿者进行的罗伊氏乳杆菌双盲安慰剂对照第一阶段安全试验 (U01AT003519)。在这项研究中，我们正在测量益生菌如何影响调节炎症的效应细胞。我们正在量化循环 T 细胞、B 细胞、单核细胞、T 细胞亚群（通过流式细胞术）。我们正在测量外周血单核细胞、粪便钙卫蛋白和粪便微生物群产生的细胞因子。该试验包括调节性 T 细胞 (Treg) 的测量，这些细胞旨在介导益生菌的免疫调节作用。因此，我们的主要目标是（在为期两年的试验中）证明罗伊氏乳杆菌对于患有绞痛的健康新生儿 (n=45) 是否安全，采用 3 种不同剂量：5 x 106、5 x 107 和 5 x 108 c.f.u。每天口服一次，持续 si 周。我们的次要目标是比较结果以确定未来随机对照试验 (RCT) 的最佳剂量。将确定以下测量的量效关系： (a) 哭泣+烦躁时间； (b) 粪便钙卫蛋白； (c) 血清细胞因子； (d) 外周血单核细胞上的Toll样受体表达； (e) 循环调节性T细胞； (f) 粪便中绞痛相关微生物（克雷伯氏菌和大肠杆菌，通过 qPCR 测定）的水平和粪便微生物群的多样性（通过 454 焦磷酸测序评估）。公众健康相关性：肠绞痛是一种严重疾病，影响约 15% 的正常婴儿，并且与产后抑郁、婚姻压力和婴儿摇晃综合症有关。我们在休斯顿研究的婴儿平均每天哭 5 个小时，并且患有与粪便中异常常驻细菌（微生物群）数量相关的肠道炎症。我们的研究将确定一种名为罗伊氏乳杆菌的益生菌（促进健康的细菌）对患有肠绞痛的正常婴儿的安全性。此外，我们将确定罗伊氏乳杆菌是否影响哭泣/烦躁时间，改变肠道炎症（粪便和血液）的测量，和/或改变常驻粪便微生物群（使用分子技术）。