Lactogenic immunity/probiotics: Effect on neonatal gut immunity

泌乳免疫/益生菌：对新生儿肠道免疫的影响

• 批准号: 7656023
• 负责人: Linda J. Saif
• 金额: $ 22.5万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7656023
• 关键词: Acute; Adhesions; Adoption; Affect; Age-Months; Anaerobic Bacteria; Animal Model; Antibiotics; Antibodies; Antibody Formation; Antigens; B-Lymphocytes; Bacteria; Bacterial Luciferases; Bifidobacterium; Binding; Biological Products; Breast Feeding; CD14 gene; Cells; Child; Colostrum; Cytotoxic T-Lymphocytes; Dendritic Cells; Derivation procedure; Developed Countries; Developing Countries; Development; Diarrhea; Disease; Dissection; Dose; Down-Regulation; Ecology; Effectiveness; Enteral; Epithelial Cells; Equilibrium; Evaluation; Exclusive Breastfeeding; Family suidae; Food; Food Hypersensitivity; Future; Gastroenteritis; Gastrointestinal Physiology; Generations; Gnotobiotic; Goals; Gram-Positive Bacteria; Health Benefit; Homeostasis; Housing; Human; Human Milk; Human poliovirus; Humoral Immunities; IL4 gene; Image; Immune; Immune response; Immune system; Immunity; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunologics; Immunomodulators; In Vitro; Incidence; Indigenous; Individual; Infant; Infant Health; Infection; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Interleukin-10; Interleukin-12; Interleukin-4; Interleukin-6; Intestinal Content; Intestinal Mucosa; Intestines; Investigation; Lactobacillus; Lactobacillus acidophilus; Lactobacillus casei rhamnosus; Licensing; Life; Link; Luc Gene; Lymphocyte; Lymphoid Tissue; Maintenance; Maternal antibody; Measures; Mediating; Mediator of activation protein; Metabolic; Methods; Microbe; Milk; Modeling; Modification; Molecular; Mothers; Mucosal Immune Responses; Mucosal Immunity; Mucous body substance; Neonatal; Oral; Oral Administration; Pathogenesis; Pattern; Polioviruses; Predisposition; Probiotics; Production; Recovery; Rehydrations; Reporter; Research; Role; Rotavirus; Rotavirus Infections; Rotavirus Vaccines; Rotavirus disease; Secretory Immunoglobulin A; Seminal; Serum; Sterility; Supplementation; Syndrome; T-Lymphocyte; Testing; Toll-like receptors; Virulent; Virus; Virus Diseases; Work; cost; cytokine; enteric pathogen; experience; feeding; gut microbiota; gut microflora; immunoregulation; imprint; improved; in vivo; innovation; insight; lactic acid bacteria; macrophage; mast cell; microorganism; monocyte; neonate; novel; novel strategies; pathogen; peripheral blood; pregnant; prevent; public health relevance; receptor; response; synergism

DESCRIPTION (provided by applicant): Breast milk contains maternal antibodies (MatAbs) that promote infant health and moderate diarrhea, and soluble immunomediators (TGF2, IL4, sCD14, etc) whose role is largely unexplored. Rotavirus (RV) is a leading cause of diarrhea in infants worldwide. Newly licensed oral RV vaccines have unproven or lower efficacies in developing countries and high costs remain obstacles to universal adoption. Although lactic acid bacteria (LAB) reduce RV diarrhea in infants, mechanisms are undefined and the effect of providing concurrent MatAb/mediators with LAB is unexplored. Our innovative studies will first assess the impact of colostrum (col)/milk and key maternal immunomediators (TGF2, IL4) on colonization by probiotics. We then determine the individual and combined effects of col/milk and LAB on immunologic maturation, moderation of RV diarrhea and homeostasis in the neonatal gut. Bifidobacteria and lactobacillus strains differ in breastfed vs formula fed infants. We hypothesize that col/milk promotes colonization by these 2 probiotic LAB, requiring fewer or lower LAB doses, and this synergism enhances gut immune maturation and homeostasis leading to moderation of rotavirus diarrhea. Preliminary studies showing that L rhamnosus GG (LGG) colonized col/milk-fed pigs long-term after a single oral dose support our novel idea. Our unique neonatal gnotobiotic (Gn) pig model is colostrum- deprived (no MatAb pre-col/milk), allowing MatAb/mediator manipulation and free of microbes (caesarean-derived, sterile housing), permitting evaluation of defined probiotics. Importantly their gastrointestinal physiology, mucosal immune responses and susceptibility to human RV diarrhea mimic that of infants. In Aim 1, we determine if col/milk containing MatAb/mediators (or TGF2+IL4) influence a) Bifidobacterium lactis and LGG colonization, persistence and distribution in the gut and b) maturation of neonatal gut immune responses. In Aim 2 we elucidate a) if col/milk with low titer RV antibody (naturally exposed sows) in concert with LAB modulates HRV pathogenesis and immunity, thereby moderating HRV diarrhea and b) if an enteropathogenic viral infection affects gut homeostasis and the LAB microflora. We will use in vivo bioluminescent imaging to study LGG interactions with/without B. lactis in the neonatal gut, and the influence of col/milk or HRV infection on these interactions. This novel approach to in vivo bacterial ecology will elucidate probiotic LAB interactions within the host. Intestinal and systemic immunologic parameters to be assessed include: 1) innate responses; 2) virus- specific, LAB-specific and total isotype B cell responses focusing on gut IgA; 3) T cell responses (Th1, Th2, Th3, Tr1 cytokines). Our innovative studies will provide improved understanding of maturation of neonatal immunity and new insights into the beneficial effects of novel col/milk MatAb/mediators in concert with probiotics in protective immunity to RV. Further, our findings will suggest new strategies to limit RV diarrhea in infants in developing countries through short-term co- administration of probiotics and col/mlk mediators to breastfed infants or pregnant mothers. This approach will promote stable LAB colonization and gut immune maturation in infants leading to disease moderation. Immunomodulation and enhancement of neonatal immune responses also have fundamental implications for tolerance to food antigens or commensals to control food allergies or inflammatory bowel syndromes as well as antibiotic-induced diarrheas. PUBLIC HEALTH RELEVANCE: Breast milk contains maternal antibodies and other soluble immunomodulators that promote infant health and moderate diarrheal diseases. Probiotics are living non-pathogenic microorganisms that, when ingested, benefit the health of their host. Because bifidobacteria and lactobacillus colonization differs between breastfed and formula-fed infants, it is important to investigate the impact of colostrum and milk on gut colonization by the probiotic strains, Bifidobacterium lactis and L. rhamnosus GG (LGG) and their combined effect on immunologic maturation of the neonatal gut. Synergy between colostrum/milk and these two probiotic lactic acid bacteria should promote immunologic maturation and maintain immunologic balance in the gut, reducing diarrheal disease (i.e. rotavirus) in infants. Rotavirus (RV) is a leading cause of diarrhea in infants worldwide, but RV vaccines fail in developing countries. We will use the neonatal gnotobiotic pig model that is colostrum-deprived, free of microbes and susceptible to human rotavirus diarrhea to evaluate the effectiveness of colostrum/milk and probiotics to prevent RV diarrhea. Our findings will suggest new strategies to limit diarrheal disease in infants in developing countries through short-term co-administration of probiotics and colostrum/milk mediators as biotherapeutic agents to breastfed infants or to pregnant mothers, leading to infant colonization. This approach will promote stable colonization by probiotics and gut immune maturation in infants resulting in disease moderation. Understanding how neonatal immune responses can be modulated and enhanced has important implications, not only for neonatal immunity to diarrheal disease, but also for induction of tolerance to food antigens and our indigenous microflora, to control food allergies or inflammatory bowel syndromes as well as antibiotic-induced diarrheas.

描述（由申请人提供）：母乳含有促进婴儿健康和中度腹泻的母体抗体（MatAbs），以及可溶性免疫介质（TGF2、IL4、sCD14等），其作用很大程度上尚未被探索。轮状病毒 (RV) 是全世界婴儿腹泻的主要原因。新获得许可的口服 RV 疫苗在发展中国家的功效尚未得到证实或较低，而且高成本仍然是普遍采用的障碍。尽管乳酸菌 (LAB) 可减少婴儿 RV 腹泻，但其机制尚不明确，并且同时提供 MatAb/介质和 LAB 的效果尚未探索。我们的创新研究将首先评估初乳 (col)/牛奶和关键母体免疫介质（TGF2、IL4）对益生菌定植的影响。然后，我们确定了 col/milk 和 LAB 对免疫成熟、RV 腹泻的缓解和新生儿肠道稳态的单独和组合影响。母乳喂养的婴儿与配方奶喂养的婴儿中的双歧杆菌和乳酸菌菌株有所不同。我们假设，大肠/牛奶会促进这 2 种益生菌 LAB 的定植，从而需要更少或更低的 LAB 剂量，并且这种协同作用可增强肠道免疫成熟和体内平衡，从而缓解轮状病毒腹泻。初步研究表明，单次口服剂量后，鼠李糖乳杆菌 GG (LGG) 在结肠/奶饲猪中长期定植，支持了我们的新想法。我们独特的新生无菌 (Gn) 猪模型不含初乳（无 MatAb 预生/牛奶），允许 MatAb/介体操作并且不含微生物（剖腹产衍生的无菌猪舍），从而允许评估特定的益生菌。重要的是，它们的胃肠道生理学、粘膜免疫反应和对人类RV腹泻的易感性与婴儿相似。在目标 1 中，我们确定含有 MatAb/介体（或 TGF2+IL4）的 col/milk 是否影响 a) 乳双歧杆菌和 LGG 在肠道中的定植、持久性和分布，以及 b) 新生儿肠道免疫反应的成熟。在目标 2 中，我们阐明 a) 含有低滴度 RV 抗体（自然暴露的母猪）的结肠/牛奶是否与 LAB 一起调节 HRV 发病机制和免疫，从而缓解 HRV 腹泻；b) 肠病性病毒感染是否影响肠道稳态和 LAB 微生物区系。我们将使用体内生物发光成像来研究新生儿肠道中 LGG 有/无乳酸双歧杆菌的相互作用，以及结肠/牛奶或 HRV 感染对这些相互作用的影响。这种体内细菌生态学的新方法将阐明益生菌 LAB 在宿主内的相互作用。待评估的肠道和全身免疫学参数包括： 1) 先天反应； 2) 病毒特异性、LAB 特异性和总同型 B 细胞反应，重点关注肠道 IgA； 3）T细胞反应（Th1、Th2、Th3、Tr1细胞因子）。我们的创新研究将加深对新生儿免疫力成熟的理解，并为新型结肠/牛奶 MatAb/介质与益生菌在 RV 保护性免疫中的有益作用提供新的见解。此外，我们的研究结果将提出新的策略，通过对母乳喂养的婴儿或孕妇短期联合施用益生菌和 col/mlk 介质来限制发展中国家婴儿的 RV 腹泻。这种方法将促进婴儿体内乳酸菌的稳定定植和肠道免疫成熟，从而缓解疾病。免疫调节和新生儿免疫反应的增强对于食物抗原或共生体的耐受性也具有根本性影响，以控制食物过敏或炎症性肠综合征以及抗生素引起的腹泻。 公共健康相关性：母乳含有母体抗体和其他可促进婴儿健康和中度腹泻疾病的可溶性免疫调节剂。益生菌是活的非病原性微生物，摄入后有益于宿主的健康。由于母乳喂养和配方奶喂养的婴儿之间的双歧杆菌和乳酸菌定植情况不同，因此研究初乳和牛奶对益生菌菌株乳双歧杆菌和鼠李糖乳杆菌 GG (LGG) 肠道定植的影响以及它们对免疫成熟的综合影响非常重要新生儿肠道。初乳/牛奶与这两种益生菌乳酸菌之间的协同作用应促进免疫成熟并维持肠道免疫平衡，减少婴儿腹泻疾病（即轮状病毒）。轮状病毒 (RV) 是全世界婴儿腹泻的主要原因，但 RV 疫苗在发展中国家无效。我们将使用缺乏初乳、不含微生物且易受人轮状病毒腹泻影响的新生无菌猪模型来评估初乳/牛奶和益生菌预防 RV 腹泻的有效性。我们的研究结果将提出新的策略，通过短期联合使用益生菌和初乳/乳介质作为母乳喂养的婴儿或怀孕母亲的生物治疗剂来限制发展中国家婴儿的腹泻病，从而导致婴儿定植。这种方法将促进婴儿体内益生菌的稳定定植和肠道免疫成熟，从而缓解疾病。了解如何调节和增强新生儿免疫反应具有重要意义，不仅对于新生儿对腹泻病的免疫力，而且对于诱导对食物抗原和我们的本土微生物群的耐受性，控制食物过敏或炎症性肠综合症以及抗生素。诱发腹泻。