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

Themilk pentasaccharide LNFPIII has therapeutic action for metabolic and autoimmune dis-eases 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 humanmilk and also known as stage-specific embryonic antigen-1. LNFPIII-NGC drives alternative activation of macrophages and dendritic cells viaNFκB 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-NGCwas completely dependent on CD14/TLR4-Ras-Raf1/TPL2-MEK axis in bonemarrow derived dendritic cells (BMDCs). In addition, LNFPIII-NGC preferen-tially induced the production of Th2 “favoring ” chemokines CCL22 andmatrix metalloprotease protein-9 in a CD14 dependent manner in BMDCs. In contrast, LNFPIII-NGC induces signifi-cantly lower levels of Th1 “favoring ” chemokines, MIP1α, MIP1β andMIP-2 compared to level