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）：母乳中含有促进婴儿健康和中度腹泻的母体抗体（MATAB），以及可溶性免疫接触者（TGF2，IL4，SCD14等），其作用在很大程度上没有探索。轮状病毒（RV）是全世界婴儿腹泻的主要原因。新许可的口服RV疫苗在发展中国家的效力未经证实或较低，高昂的成本仍然是普遍采用的障碍。尽管乳酸细菌（LAB）降低了婴儿的RV腹泻，但机制是不确定的，并且未探索了同时使用MATAB/介体的效果。我们的创新研究将首先评估初乳（Col）/牛奶和关键母体免疫接种者（TGF2，IL4）对益生菌定植的影响。然后，我们确定Col/Milk和Lab对新生儿肠道中免疫学成熟，RV腹泻和稳态的适度的个体和组合作用。双歧杆菌和乳酸杆菌菌株在母乳喂养的婴儿与配方奶粉中有所不同。我们假设Col/牛奶促进了这2个益生菌实验室的定殖，需要更少或更低的实验室剂量，这种协同作用增强了肠道免疫成熟和稳态，导致轮状病毒腹泻的适度。初步研究表明，L Rhamnosus gg（LGG）在单个口服剂量后长期定植了Col/牛奶喂养的猪，这支持了我们的新思想。我们独特的新生儿gnotobiotic（GN）猪模型被剥夺了初乳（无MATAB pre-Col/牛奶），可以使MATAB/介体操纵和不含微生物（剖腹产的无菌住房），允许评估已定义的益生菌。重要的是，它们的胃肠道生理，粘膜免疫反应和对人RV腹泻的敏感性模仿婴儿。在AIM 1中，我们确定含有MATAB/介质的Col/牛奶（或TGF2+IL4）是否影响a）乳酸杆菌和LGG定植，肠道和b）新生儿肠道免疫反应的成熟。在AIM 2中，我们阐明a）如果与实验室调节的col/牛奶，如果具有低滴度RV抗体（自然暴露的母猪），则可以调节HRV的发病机理和免疫力，从而调节HRV腹泻和b）如果肠病毒病毒感染会影响Gute脑稳态和Lab Microflora。我们将使用体内生物发光成像来研究新生儿肠道中与乳酸乳乳杆菌的LGG相互作用，以及Col/Milk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宿主内的益生菌实验室相互作用。要评估的肠道和全身免疫学参数包括：1）先天反应； 2）针对肠道IgA的病毒特异性，实验室特异性和总同种型B细胞反应； 3）T细胞反应（TH1，TH2，TH3，TR1细胞因子）。我们的创新研究将提高人们对新生儿免疫的成熟以及对新型Col/牛奶MATAB/介体的有益作用的新见解，其中包括对RV的保护性免疫的益生菌。此外，我们的发现将提出新的策略，以限制发展中国家的婴儿腹泻，通过短期益生菌和Col/MLK介体的短期共同给予母乳喂养的婴儿或怀孕的母亲。这种方法将促进婴儿稳定的实验室定植和肠道免疫成熟，从而导致疾病节制。新生儿免疫反应的免疫调节和增强也对耐受食物抗原或共生的耐受性具有根本性的影响，以控制食物过敏或炎症性肠综合征以及抗生素诱导的腹泻。 公共卫生相关性：母乳含有孕妇抗体和其他可溶性免疫调节剂，可促进婴儿健康和中度腹泻疾病。益生菌是生命的非致病微生物，当摄入时，益生菌会受益于其宿主的健康。由于母乳喂养和配方蛋白喂养的婴儿之间的双歧杆菌和乳酸菌定植不同，因此重要的是研究乳酸菌菌株，乳酸乳酸菌和rhamnosus l. rhamnosus gg（LGG）及其对免疫学的综合作用的益生菌菌株，乳酸菌和l. rhamnosus l.初乳/牛奶与这两种益生菌乳酸细菌之间的协同作用应促进免疫学成熟并维持肠道中的免疫平衡，从而减少婴儿的腹泻病（即轮状病毒）。轮状病毒（RV）是全世界婴儿腹泻的主要原因，但发展中国家RV疫苗未能进行。我们将使用剥夺初乳的新生儿gnotobiotic猪模型，不含微生物并受到人轮状病毒腹泻易感性，以评估初乳/牛奶和益生菌的有效性以防止RV腹泻。我们的发现将提出新的策略，通过短期共同给予益生菌和初乳/牛奶介质作为母乳喂养的婴儿或怀孕的母亲的生物治疗剂，从而限制发展中国家腹泻病的新策略，从而导致婴儿定植。这种方法将促进益生菌和肠道免疫成熟的稳定定殖，导致疾病适度。了解新生儿免疫反应如何调节和增强具有重要意义，这不仅是对新生儿对腹泻病的免疫力的影响，还为了诱导对食物抗原和我们的本地微生物的耐受性，可以控制食物过敏或炎性肠综合症以及抗生素诱导的糖尿病诱导的疾病。