Anti-inflammatory effects of Bifidobacterium longum subsp infantis secretions on fetal human enterocytes are mediated by TLR-4 receptors.

The therapeutic and preventive application of probiotics for necrotizing enterocolitis (NEC) has been supported by more and more experimental and clinical evidence in which Toll-like receptor 4 (TLR-4) exerts a significant role. In immune cells, probiotics not only regulate the expression of TLR-4 but also use the TLR-4 to modulate the immune response. Probiotics may also use the TLR-4 in immature enterocytes for anti-inflammation. Here we demonstrate that probiotic conditioned media (PCM) from Bifidobacterium longum supp infantis but not isolated organisms attenuates interleukin-6 (IL-6) induction in response to IL-1β by using TLR-4 in a human fetal small intestinal epithelial cell line (H4 cells), human fetal small intestinal xenografts, mouse fetal small intestinal organ culture tissues, and primary NEC enterocytes. Furthermore, we show that PCM, using TLR-4, downregulates the mRNA expression of interleukin-1 receptor-associated kinase 2 (IRAK-2), a common adapter protein shared by IL-1β and TLR-4 signaling. PCM also reduces the phosphorylation of the activator-protein 1 (AP-1) transcription factors c-Jun and c-Fos in response to IL-1β stimulation in a TLR-4-dependent manner. This study suggests that PCM may use TLR-4 through IRAK-2 and via AP-1 to prevent IL-1β-induced IL-6 induction in immature enterocytes. Based on these observations, the combined use of probiotics and anti-TLR-4 therapy to prevent NEC may not be a good strategy.

[1]  C. Cahill,et al.  Differential Expression of the Activator Protein 1 Transcription Factor Regulates Interleukin-1ß Induction of Interleukin 6 in the Developing Enterocyte , 2016, PloS one.

[2]  P. Brown,et al.  TLR4 has a TP53-dependent dual role in regulating breast cancer cell growth , 2015, Proceedings of the National Academy of Sciences.

[3]  W. Walker,et al.  Secreted Metabolites of Bifidobacterium infantis and Lactobacillus acidophilus Protect Immature Human Enterocytes from IL-1β-Induced Inflammation: A Transcription Profiling Analysis , 2015, PloS one.

[4]  G. Perdigón,et al.  Stimulation of Innate Immune Cells Induced by Probiotics: Participation of Toll-Like Receptors , 2015 .

[5]  W. Walker,et al.  The Toll-like receptor-4 in human and mouse colonic epithelium is developmentally regulated: a possible role in Necrotizing Enterocolitis , 2014, Pediatric Research.

[6]  K. Kalanetra,et al.  Bifidobacterium longum subsp. infantis in experimental necrotizing enterocolitis: alterations in inflammation, innate immune response, and the microbiota , 2014, Pediatric Research.

[7]  A. Castro,et al.  Partial inhibition of Cdk1 in G2 phase overrides the SAC and decouples mitotic events , 2014, Cell cycle.

[8]  D. Hackam,et al.  Toll-like receptor regulation of intestinal development and inflammation in the pathogenesis of necrotizing enterocolitis. , 2014, Pathophysiology : the official journal of the International Society for Pathophysiology.

[9]  H. Kitazawa,et al.  Modulation of Intestinal TLR4-Inflammatory Signaling Pathways by Probiotic Microorganisms: Lessons Learned from Lactobacillus jensenii TL2937 , 2014, Front. Immunol..

[10]  A. Afrazi,et al.  Innate Immune Signaling in the Pathogenesis of Necrotizing Enterocolitis , 2013, Clinical & developmental immunology.

[11]  S. Rautava,et al.  Probiotics prevent necrotizing enterocolitis by modulating enterocyte genes that regulate innate immune-mediated inflammation. , 2013, American journal of physiology. Gastrointestinal and liver physiology.

[12]  A. Kraneveld,et al.  Dual Role of Toll-Like Receptors in Asthma and Chronic Obstructive Pulmonary Disease , 2012, Pharmacological Reviews.

[13]  J. M. Rhoads,et al.  Lactobacillus reuteri strains reduce incidence and severity of experimental necrotizing enterocolitis via modulation of TLR4 and NF-κB signaling in the intestine. , 2012, American journal of physiology. Gastrointestinal and liver physiology.

[14]  E. Pamer,et al.  Innate immune signaling in defense against intestinal microbes , 2012, Immunological reviews.

[15]  L. Joosten,et al.  The Evolutionary History of TLR4 Polymorphisms in Europe , 2011, Journal of Innate Immunity.

[16]  R. Uauy,et al.  The Mechanism of Excessive Intestinal Inflammation in Necrotizing Enterocolitis: An Immature Innate Immune Response , 2011, PloS one.

[17]  S. Akira,et al.  Pattern Recognition Receptors and Inflammation , 2010, Cell.

[18]  Axel Weber,et al.  Interleukin-1 (IL-1) Pathway , 2010, Science Signaling.

[19]  D. Bassler,et al.  Probiotics Reduce the Risk of Necrotizing Enterocolitis in Preterm Infants: A Meta-Analysis , 2009, Neonatology.

[20]  C. Van Ginneken,et al.  Diet-dependent Mucosal Colonization and Interleukin-1β Responses in Preterm Pigs Susceptible to Necrotizing Enterocolitis , 2009, Journal of pediatric gastroenterology and nutrition.

[21]  T. Mogensen.  Pathogen Recognition and Inflammatory Signaling in Innate Immune Defenses , 2009, Clinical Microbiology Reviews.

[22]  Elizabeth E. Molnar,et al.  TRAF6 Autoubiquitination-Independent Activation of the NFκB and MAPK Pathways in Response to IL-1 and RANKL , 2008, PloS one.

[23]  Osamu Takeuchi,et al.  Sequential control of Toll-like receptor–dependent responses by IRAK1 and IRAK2 , 2008, Nature Immunology.

[24]  D. Hackam,et al.  The role of epithelial Toll‐like receptor signaling in the pathogenesis of intestinal inflammation , 2008, Journal of leukocyte biology.

[25]  A. Bowie,et al.  IRAK-2 Participates in Multiple Toll-like Receptor Signaling Pathways to NFκB via Activation of TRAF6 Ubiquitination* , 2007, Journal of Biological Chemistry.

[26]  T. Raju,et al.  New Therapies and Preventive Approaches for Necrotizing Enterocolitis: Report of a Research Planning Workshop , 2007, Pediatric Research.

[27]  R. Sartor,et al.  Bacterial symbionts induce a FUT2-dependent fucosylated niche on colonic epithelium via ERK and JNK signaling. , 2007, American journal of physiology. Gastrointestinal and liver physiology.

[28]  D. Hackam,et al.  A Critical Role for TLR4 in the Pathogenesis of Necrotizing Enterocolitis by Modulating Intestinal Injury and Repair1 , 2007, The Journal of Immunology.

[29]  R. Wait,et al.  Mass Spectrometric Analysis of the Endogenous Type I Interleukin-1 (IL-1) Receptor Signaling Complex Formed after IL-1 Binding Identifies IL-1RAcP, MyD88, and IRAK-4 as the Stable Components* , 2007, Molecular & Cellular Proteomics.

[30]  W. Walker,et al.  Secreted Probiotic Factors Ameliorate Enteropathogenic Infection in Zinc-Deficient Human Caco-2 and T84 Cell Lines , 2007, Pediatric Research.

[31]  B. Evers,et al.  Effectiveness of siRNA uptake in target tissues by various delivery methods. , 2007, Surgery.

[32]  W. Walker,et al.  Glucocorticoid responsiveness in developing human intestine: possible role in prevention of necrotizing enterocolitis. , 2005, American journal of physiology. Gastrointestinal and liver physiology.

[33]  W. Falk,et al.  The Interleukin 1 (IL-1) Receptor Accessory Protein Toll/IL-1 Receptor Domain , 2003, Journal of Biological Chemistry.

[34]  W. Walker,et al.  Normal and glucocorticoid-induced development of the human small intestinal xenograft. , 2003, American journal of physiology. Regulatory, integrative and comparative physiology.

[35]  Michael Levine,et al.  Linear signaling in the Toll-Dorsal pathway of Drosophila: activated Pelle kinase specifies all threshold outputs of gene expression while the bHLH protein Twist specifies a subset. , 2002, Development.

[36]  Holger Wesche,et al.  IRAK-4: A novel member of the IRAK family with the properties of an IRAK-kinase , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[37]  M. Baldeón,et al.  Evidence for an Innate Immune Response in the Immature Human Intestine: Toll-Like Receptors on Fetal Enterocytes , 2001, Pediatric Research.

[38]  Shuguang Wu,et al.  IRAK-2 and PI 3-Kinase Synergistically Activate NF-κB and AP-1 , 2000, Inflammation.

[39]  W. Walker,et al.  Inflammation in the developing human intestine: A possible pathophysiologic contribution to necrotizing enterocolitis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[40]  A. Hoyos.  Reduced incidence of necrotizing enterocolitis associated with enteral administration of Lactobacillus acidophilus and Bifidobacterium infantis to neonates in an intensive care unit. , 1999, International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases.

[41]  J. Hasday,et al.  Inflammatory cytokine mRNAs in surgical specimens of necrotizing enterocolitis and normal newborn intestine. , 1997, Pediatric pathology & laboratory medicine : journal of the Society for Pediatric Pathology, affiliated with the International Paediatric Pathology Association.

[42]  Zhaodan Cao,et al.  TRAF6 is a signal transducer for interleukin-1 , 1996, Nature.

[43]  R. Ezzell,et al.  Human fetal enterocytes in vitro: modulation of the phenotype by extracellular matrix. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Z. Cao,et al.  IRAK: A Kinase Associated with the Interleukin-1 Receptor , 1996, Science.

[45]  M. Henry,et al.  Necrotizing Enterocolitis , 1977, Pediatric Surgery.

[46]  C. Lebrilla,et al.  Bifidobacterium longum subspecies infantis: champion colonizer of the infant gut , 2015, Pediatric Research.

[47]  D. Bassler,et al.  Probiotics for prevention of necrotizing enterocolitis in preterm infants (Review) , 2014 .

[48]  D. Ye,et al.  Epithelial and Mesenchymal Cell Biology IL-1-Induced Increase in Intestinal Epithelial Tight Junction Permeability Is Mediated by MEKK-1 Activation of Canonical NF-B Pathway , 2010 .

[49]  D. Bassler,et al.  Probiotics for prevention of necrotizing enterocolitis in preterm infants. , 2008, The Cochrane database of systematic reviews.

[50]  W. Falk,et al.  The Interleukin 1 (IL-1) Receptor Accessory Protein Toll/IL-1 Receptor Domain ANALYSIS OF PUTATIVE INTERACTION SITES BY IN VITRO MUTAGENESIS AND MOLECULAR MODELING* , 2003 .

[51]  M. Rehli,et al.  Evolution of the TIR, tolls and TLRs: functional inferences from computational biology. , 2002, Current topics in microbiology and immunology.

[52]  F. Guo,et al.  IRAK-2 and PI 3-kinase synergistically activate NF-kappaB and AP-1. , 2000, Inflammation.