Immune Netw.  2017 Aug;17(4):228-236. 10.4110/in.2017.17.4.228.

Yersinia enterocolitica Exploits Signal Crosstalk between Complement 5a Receptor and Toll-like Receptor 1/2 and 4 to Avoid the Bacterial Clearance in M cells

• Kim SH
• Jang YS

• KMID: 2388077
• DOI: http://doi.org/10.4110/in.2017.17.4.228

Abstract

In the intestinal mucosal surface, microfold cells (M cells) are the representative gateway for the uptake of luminal antigens. At the same time, M cells are the primary infection site for pathogens invading mucosal surface for their infection. Although it is well recognized that many mucosal pathogens exploit the M cells for their infection, the mechanism to infect M cells utilized by pathogens is not clearly understood yet. In this study, we found that M cells expressing complement 5a (C5a) receptor (C5aR) also express Toll-like receptor (TLR) 1/2 and TLR4. Infection of Yersinia enterocolitica, an M cell-invading pathogen, synergistically regulated cyclic adenosine monophosphate-dependent protein kinase A (cAMP-PKA) signaling which are involved in signal crosstalk between C5aR and TLRs. In addition, Y. enterocolitica infection into M cells was enhanced by C5a treatment and this enhancement was abrogated by C5a antagonist treatment. Finally, Y. enterocolitica infection into M cells was unsuccessful in C5aR knock-out mice. Collectively, we suggest that exploit the crosstalk between C5aR and TLR signaling is one of infection mechanisms utilized by mucosal pathogens to infect M cells.

Keyword

Complement 5a receptor; Immune evade; M cell; Toll-like receptor; Yersinia enterocolitica

MeSH Terms

Adenosine
Animals
Complement C5a*
Complement System Proteins*
Cyclic AMP-Dependent Protein Kinases
Mice
Mice, Knockout
Phenobarbital
Receptor, Anaphylatoxin C5a*
Toll-Like Receptors*
Yersinia enterocolitica*
Yersinia*
Adenosine
Complement C5a
Complement System Proteins
Cyclic AMP-Dependent Protein Kinases
Phenobarbital
Receptor, Anaphylatoxin C5a
Toll-Like Receptors

Figure

• 

  Figure 1 Y. enterocolitica interacted with C5aR and TLR1/2 in PP M cells. (A, B) Whole-mounted PPs were stained with either NKM 16-2-4 anti-M cell Ab (A) or anti-GP2 Ab (B), anti-C5aR Ab, either anti-TLR4 Ab (A) or anti-TLR1/2 Ab (B), and WGA. (C, D) PPs were prepared at 10 min after oral administration of CFSE-labeled Y. enterocolitica and stained with anti-p-C5aR Ab, anti-TLR1/2 Ab, and WGA. The samples were analyzed by confocal laser scanning microscopy. (C) and (D) show stacked images of XY and XZ images. The dotted lines depict M cells. Scale bars represent 20 μm. WGA, wheat-germ agglutinin.

• 

  Figure 2 The expression level of cAMP signaling-related genes increased in human M-like cells incubated with Y. enterocolitica. The human M-like cells were differentiated from Caco-2 cells by co-culturing with Raji. After stimulating the cells with either C5a together with LPS, or Y. enterocolitica for 6 h, change in the gene expression was evaluated through quantitative real-time PCR. The results from 3 independent experiments are presented as a fold regulation which is the negative inverse of the fold change if the fold change is less than 1. N.S., not significant. **p<0.01 and ***p<0.001 indicate significant differences between the groups compared.

• 

  Figure 3 Y. enterocolitica induces the activation of PKA in PP M cells. The PPs were prepared from the mice orally administered with or without CFSE-labeled Y. enterocolitica, stained with anti-GP2 Ab and anti-p-PKA substrate Ab, and counter stained with DAPI. The samples were analyzed by confocal laser scanning microscopy. The image is a stack of XY. The arrows indicate M cells. Scale bars represent 20 μm.

• 

  Figure 4 Y. enterocolitica exploits C5aR signaling for their survival in vivo. (A) The human M-like cells and E-like cells were either pretreated or not with C5aRA and then infected with Y. enterocolitica. (B, C) The WT mice and C5aR KO mice were orally administered with Y. enterocolitica. The M-like cells (A), PP cells (B), or MLN cells (C) were prepared 3 h after infection and used to determine viable Y. enterocolitica CFU. Data are represented as mean±SE (n=3) in the group. CFU, colony-forming unit; SE, standard error; ND, not determined. *p<0.05, **p<0.01, and ***p<0.001 indicate significant differences between the groups compared.

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