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

• 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
• 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

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）：绞痛是一个严重的问题，其特征是每天哭泣，哭泣和尖叫超过3个小时，并且在生命的头三个月中影响了约15％的正常婴儿。我们最近发表了定义绞痛婴儿的关键临床和微生物特征的试验。与正常婴儿相比，患有绞痛的婴儿 表现出哭泣时间增加了四倍，在粪便钙染色蛋白的水平（肠道炎症度量）中增加了一倍，以及粪便菌群的变化，其特征是细菌多样性降低和klebsiella菌患病率的升高。这些结果表明了一个新的假设：婴儿哭泣是因为肠子发炎，这可能是由于微生物群异常而导致的。我们的调查将集中在绞痛的健康婴儿中，以显示益生菌的安全性，该益生菌已显示出可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的新生大鼠模型中，对肠乳杆菌的喂养降低了肠道炎症，死亡率降低了50％。目前，我们接近成人志愿者L. Reuteri的双盲安慰剂对照的1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细胞子集（通过流式细胞仪）。我们正在测量外周血单核细胞，粪便钙染色素和粪便菌群的细胞因子产生。该试验包括测量调节性T细胞（TREG），该细胞提议介导益生菌的免疫调节作用。因此，我们的主要目的是（在为期两年的试验中），如果Reuteri在具有绞痛的健康新生儿（n = 45）的健康新生儿中是安全的，则以3种不同的剂量：5 x 106、5 x 107和5 x 108 c.f.u.每天通过口嘴一次，持续一周。我们的次要目的是比较结果，以确定未来随机对照试验（RCT）的最佳剂量。将确定以下措施的剂量效应关系：（a）哭泣 +大惊小怪的时间； （b）粪便钙蛋白酶； （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T细胞； （f）粪便相关生物的粪便水平（克雷伯菌和大肠杆菌，由qPCR确定）和粪便菌群的多样性（通过454 pyrosequencing评估）。 公共卫生相关性：绞痛是一种严重的疾病，影响了约15％的正常婴儿，并且与产后抑郁症，婚姻压力和动摇婴儿综合征有关。我们在休斯顿学习的婴儿平均每天哭了5个小时，并且与粪便中常数细菌（Microbobiota）异常的肠道炎症有关。我们的研究将确定在正常婴儿患有绞痛的婴儿中称为乳酸杆菌的益生菌（促进健康的细菌）的安全性。此外，我们将确定Reuteri是否会影响哭泣/烦恼的时间，改变肠道炎症的测量（在粪便和血液中）和/或改变驻留的粪便微生物群（使用分子技术）。