Toll-like receptor 2 is critical for induction of Reg3 beta expression and intestinal clearance of Yersinia pseudotuberculosis.
OBJECTIVE
Yersinia pseudotuberculosis causes ileitis and mesenteric lymphadenitis by mainly invading the Peyer's patches that are positioned in the terminal ileum. Whereas toll-like-receptor 2 (TLR2) controls mucosal inflammation by detecting certain microbiota-derived signals, its exact role in protecting Peyer's patches against bacterial invasion has not been defined.


METHODS
Wild-type, Tlr2-, Nod2- and MyD88-deficient animals were challenged by Y pseudotuberculosis via the oral or systemic route. The role of microbiota in conditioning Peyer's patches against Yersinia through TLR2 was assessed by delivering, ad libitum, exogenous TLR2 agonists in drinking water to germ-free and streptomycin-treated animals. Bacterial eradication from Peyer's patches was measured by using a colony-forming unit assay. Expression of cryptdins and the c-type lectin Reg3 beta was quantified by quantitative reverse transcriptase polymerase chain reaction analysis.


RESULTS
Our data demonstrated that Tlr2-deficient mice failed to limit Yersinia dissemination from the Peyer's patches and succumbed to sepsis independently of nucleotide-binding and oligomerisation domain 2 (NOD2). Recognition of both microbiota-derived and myeloid differentiation factor 88 (MyD88)-mediated elicitors was found to be critically involved in gut protection against Yersinia-induced lethality, while TLR2 was dispensable to systemic Yersinia infection. Gene expression analyses revealed that optimal epithelial transcript level of the anti-infective Reg3 beta requires TLR2 activation. Consistently, Yersinia infection triggered TLR2-dependent Reg3 beta expression in Peyer's patches. Importantly, oral treatment with exogenous TLR2 agonists in germ-free animals was able to further enhance Yersinia-induced expression of Reg3 beta and to restore intestinal resistance to Yersinia. Lastly, genetic ablation of Reg3 beta resulted in impaired clearance of the bacterial load in Peyer's patches.


CONCLUSIONS
TLR2/REG3 beta is thus an essential component in conditioning epithelial defence signalling pathways against bacterial invasion.
Dessein
R
R
Inserm, U801, Université de Lille 2, Institut Pasteur de Lille, Lille, France.
Gironella
M
M
Vignal
C
C
Peyrin-Biroulet
L
L
Sokol
H
H
Secher
T
T
Lacas-Gervais
S
S
Gratadoux
J-J
JJ
Lafont
F
F
Dagorn
J-C
JC
Ryffel
B
B
Akira
S
S
Langella
P
P
Nùñez
G
G
Sirard
J-C
JC
Iovanna
J
J
Simonet
M
M
Chamaillard
M
M
D000818:Animals; D002460:Cell Line; D004847:Epithelial Cells / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D017353:Gene Deletion; D020869:Gene Expression Profiling / Q000379:methods; D005856:Germ-Free Life; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D046529:Microscopy, Electron, Transmission; D053473:Nod2 Signaling Adaptor Protein / Q000235:genetics / Q000378:metabolism; D000073718:Pancreatitis-Associated Proteins; D010581:Peyer's Patches / Q000378:metabolism / Q000382:microbiology* / Q000648:ultrastructure; D011506:Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015398:Signal Transduction / Q000502:physiology*; D051195:Toll-Like Receptor 2 / Q000235:genetics / Q000378:metabolism*; D015011:Yersinia pseudotuberculosis; D015012:Yersinia pseudotuberculosis Infections / Q000378:metabolism*
D053473:Nod2 Signaling Adaptor Protein; C495945:Nod2 protein, mouse; D000073718:Pancreatitis-Associated Proteins; D011506:Proteins; C490253:Reg3b protein, mouse; D051195:Toll-Like Receptor 2