A Neoglycoconjugate Containing the Human Milk Sugar LNFPIII Drives Anti-Inflammatory Activation of Antigen Presenting Cells in a CD14 Dependent Pathway.

Abstract

The milk pentasaccharide LNFPIII has therapeutic action for metabolic and autoimmune diseases and prolongs transplant survival in mice when presented as a neoglycoconjugate. Within LNFPIII is the Lewisx trisaccharide, expressed by many helminth parasites. In humans, LNFPIII is found in human milk and also known as stage-specific embryonic antigen-1. LNFPIII-NGC drives alternative activation of macrophages and dendritic cells via NFκB activation in a TLR4 dependent mechanism. However, the connection between LNFPIII-NGC activation of APCs, TLR4 signaling and subsequent MAP kinase signaling leading to anti-inflammatory activation of APCs remains unknown. In this study we determined that the innate receptor CD14 was essential for LNFPIII-NGC induction of both ERK and NFkB activation in APCs. Induction of ERK activation by LNFPIII-NGC was completely dependent on CD14/TLR4-Ras-Raf1/TPL2-MEK axis in bone marrow derived dendritic cells (BMDCs). In addition, LNFPIII-NGC preferentially induced the production of Th2 "favoring" chemokines CCL22 and matrix metalloprotease protein-9 in a CD14 dependent manner in BMDCs. In contrast, LNFPIII-NGC induces significantly lower levels of Th1 "favoring" chemokines, MIP1α, MIP1β and MIP-2 compared to levels in LPS stimulated cells. Interestingly, NGC of the identical human milk sugar LNnT, minus the alpha 1-3 linked fucose, failed to activate APCs via TLR4/MD2/CD14 receptor complex, suggesting that the alpha 1-3 linked fucose in LNFPIII and not on LNnT, is required for this process. Using specific chemical inhibitors of the MAPK pathway, we found that LNFPIII-NGC induction of CCL22, MMP9 and IL-10 production was dependent on ERK activation. Over all, this study suggests that LNFPIII-NGC utilizes CD14/TLR4-MAPK (ERK) axis in modulating APC activation to produce anti-inflammatory chemokines and cytokines in a manner distinct from that seen for the pro-inflammatory PAMP LPS. These pathways may explain the in vivo therapeutic effect of LNFPIII-NGC treatment for inflammation based diseases.

• Authors: Smanla Tundup, Leena Srivastava, Thomas Norberg, Wendy Watford, Donald Harn
• Journal: PloS one (PLoS One) Vol. 10 Issue 9 Pg. e0137495 ( 2015) ISSN: 1932-6203 [Electronic] United States
• PMID: 26340260 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Amino Sugars
• Anti-Inflammatory Agents
• Ccl22 protein, mouse
• Ccl3 protein, mouse
• Ccl4 protein, mouse
• Chemokine CCL22
• Chemokine CCL3
• Chemokine CCL4
• Chemokine CXCL2
• Cxcl2 protein, mouse
• Glycoconjugates
• Lipopolysaccharide Receptors
• NF-kappa B
• Polysaccharides
• Proto-Oncogene Proteins
• Tlr4 protein, mouse
• Toll-Like Receptor 4
• lacto-N-fucopentaose III
• Proto-Oncogene Proteins c-raf
• Extracellular Signal-Regulated MAP Kinases
• MAP Kinase Kinase Kinases
• Map3k8 protein, mouse
• Matrix Metalloproteinase 9

Topics

• Amino Sugars (chemistry, pharmacology)
• Animals
• Anti-Inflammatory Agents (chemistry, pharmacology)
• Bone Marrow Cells (cytology, drug effects, immunology)
• Cell Line
• Chemokine CCL22 (genetics, immunology)
• Chemokine CCL3 (genetics, immunology)
• Chemokine CCL4 (genetics, immunology)
• Chemokine CXCL2 (genetics, immunology)
• Dendritic Cells (cytology, drug effects, immunology)
• Extracellular Signal-Regulated MAP Kinases (genetics, immunology)
• Gene Expression Regulation
• Glycoconjugates (chemistry, pharmacology)
• Humans
• Lipopolysaccharide Receptors (genetics, immunology)
• MAP Kinase Kinase Kinases (genetics, immunology)
• Macrophages (cytology, drug effects, immunology)
• Matrix Metalloproteinase 9 (genetics, immunology)
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• NF-kappa B (genetics, immunology)
• Polysaccharides (chemistry, pharmacology)
• Primary Cell Culture
• Proto-Oncogene Proteins (genetics, immunology)
• Proto-Oncogene Proteins c-raf (genetics, immunology)
• Signal Transduction
• Th1-Th2 Balance (drug effects)
• Toll-Like Receptor 4 (genetics, immunology)