PMID:24409285

Citation	Sathe, A and Reddy, KV (2014) TLR9 and RIG-I Signaling in Human Endocervical Epithelial Cells Modulates Inflammatory Responses of Macrophages and Dendritic Cells In Vitro. PLoS ONE 9:e83882
Abstract	The innate immune system has evolved to recognize invading pathogens through pattern recognition receptors (PRRs).Among PRRs, Toll like receptors (TLRs 3, 7/8,9) and RIG-I like receptors (RLRs) have been shown to recognize viral components. Mucosal immune responses to viral infections require coordinated actions from epithelial as well as immune cells. In this respect, endocervical epithelial cells (EEC's) play an important role in initiating innate immune responses via PRRs. It is unknown whether EEC's can alter immune responses of macrophages and dendritic cells (DC's) like its counterparts in intestinal and respiratory systems. In this study, we show that endocervical epithelial cells (End1/E6E7) express two key receptors, TLR9 and RIG-I involved in anti-viral immunity. Stimulation of End1/E6E7 cells lead to the activation of NF-κB and increased secretion of pro-inflammatory cytokines, IL-6 and IL-8. Polarized End1/E6E7 cells responded to apical stimulation with ligands of TLR9 and RIG-I, CpG-ODN and Poly(I:C)LL respectively, without compromising End1/E6E7 cell integrity. At steady state, spent medium from End1/E6E7 cells significantly reduced secretion of pro-inflammatory cytokines from LPS treated human primary monocyte derived macrophages (MDMs) and DC:T cell co-cultures. Spent medium from End1/E6E7 cells stimulated with ligands of TLR9/RIG-I restored secretion of pro-inflammatory cytokines as well as enhanced phagocytosis and chemotaxis of monocytic U937 cells. Spent medium from CpG-ODN and Poly(I:C)LL stimulated End1/E6E7 cells showed significant increased secretion of IL-12p70 from DC:T cell co-cultures. The anti-inflammatory effect of spent media of End1/E6E7 cell was observed to be TGF-β dependent. In summary, the results of our study indicate that EEC's play an indispensable role in modulating anti-viral immune responses at the female lower genital tract.
Links	PubMed PMC3883652 Online version:10.1371/journal.pone.0083882
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HUMAN:DDX58	involved_in	GO:0032757: positive regulation of interleukin-8 production	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:DDX58	involved_in	GO:0032755: positive regulation of interleukin-6 production	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:DDX58	involved_in	GO:0010628: positive regulation of gene expression	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:DDX58	involved_in	GO:0032725: positive regulation of granulocyte macrophage colony-stimulating factor production	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:DDX58		GO:0032755: positive regulation of interleukin-6 production	ECO:0000314:		P		In Figure 2A agonists of RIG-I poly LL and CpG-ODN show that RIG-I increases the production of Interleukin 6 in End1/E6E7 cells as shown by ELISA test.	complete CACAO 9273
HUMAN:DDX58		GO:0032757: positive regulation of interleukin-8 production	ECO:0000314:		P		In Figure 2A agonists of RIG-I poly LL and CpG-ODN show that RIG-I increases the production of Interleukin 8 in End1/E6E7 cells as shown by ELISA test	complete CACAO 9274
HUMAN:DDX58		GO:0010628: positive regulation of gene expression	ECO:0000314:		P		Figure 3 A graph 1 and Figure 3b show that when RIG-I agonists are applied there is an increased expression of p65-NF-kB when compared to the control thus showingthat RIG-I increases expression of p65 NF-kB in End1/E6E7 cells.	complete CACAO 9277
HUMAN:DDX58		GO:0032725: positive regulation of granulocyte macrophage colony-stimulating factor production	ECO:0000314:		P		Figure 4 A and B show that when subjected to agonists RIG-I increases the production of GM-CSF in End1/E6E7 cells when compared to the control.	complete CACAO 9284
HUMAN:TLR9		GO:0032755: positive regulation of interleukin-6 production	ECO:0000314:		P		In figure 2A both CpG-ODN and Poly LL are agonists of TLR9 and when added increase the amount of Interleukin 6 produced in End1/E6E7 cells.	complete CACAO 9269
HUMAN:TLR9		GO:0032757: positive regulation of interleukin-8 production	ECO:0000314:		P		In figure 2A both CpG-ODN and Poly LL are agonists of TLR9 and when added increase the amount of Interleukin 8 produced in End1/E6E7 cells.	complete CACAO 9271
HUMAN:TLR9		GO:0010628: positive regulation of gene expression	ECO:0000314:		P		In figure 3A graph 1 and figure 3B show that when compared to the control, cells that are subjected to agnoists of TLR9 show an increased expression of p65-NF-kB in End1/E6E7 cells. This suggests that TLR9 activation leads to increased expression of P65-NF-kB.	complete CACAO 9278
HUMAN:TLR9		GO:0032725: positive regulation of granulocyte macrophage colony-stimulating factor production	ECO:0000314:		P		Figure 4 A and B show that when subjected to agonists TLR9 increases the production of GM-CSF in End1/E6E7 cells when compared to the control.	complete CACAO 9279
HUMAN:TLR9	involved_in	GO:0010628: positive regulation of gene expression	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:TLR9	involved_in	GO:0032755: positive regulation of interleukin-6 production	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:TLR9	involved_in	GO:0032725: positive regulation of granulocyte macrophage colony-stimulating factor production	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
HUMAN:TLR9	involved_in	GO:0032757: positive regulation of interleukin-8 production	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete

References

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