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Interaction of fibrinogen and muramidase-released protein promotes the development of Streptococcus suis meningitis.

Wang J, Kong D, Zhang S, Jiang H, Zheng Y, Zang Y, Hao H, Jiang Y - Front Microbiol (2015)

Bottom Line: Muramidase-released protein (MRP) is as an important virulence marker of Streptococcus suis (S. suis) serotype 2.In this study, we found that the deletion of mrp significantly impairs the hFg-mediated adherence and traversal ability of S. suis across human cerebral microvascular endothelial cells (hCMEC/D3).Measurement of the permeability to Lucifer yellow in vitro and Evans blue extravasation in vivo show that the MRP-hFg interaction significantly increases the permeability of the blood-brain barrier (BBB).

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences , Beijing, China ; Urumqi Ethnic Cadres' College , Urumqi, China.

ABSTRACT
Muramidase-released protein (MRP) is as an important virulence marker of Streptococcus suis (S. suis) serotype 2. Our previous works have shown that MRP can bind human fibrinogen (hFg); however, the function of this interaction in S. suis meningitis is not known. In this study, we found that the deletion of mrp significantly impairs the hFg-mediated adherence and traversal ability of S. suis across human cerebral microvascular endothelial cells (hCMEC/D3). Measurement of the permeability to Lucifer yellow in vitro and Evans blue extravasation in vivo show that the MRP-hFg interaction significantly increases the permeability of the blood-brain barrier (BBB). In the mouse meningitis model, wild type S. suis caused higher bacterial loads in the brain and more severe histopathological signs of meningitis than the mrp mutant at day 3 post-infection. Western blot analysis and immunofluorescence observations reveal that the MRP-hFg interaction can destroy the cell adherens junction protein p120-catenin of hCMEC/D3. These results indicate that the MRP-hFg interaction is important in the development of S. suis meningitis.

No MeSH data available.


Related in: MedlinePlus

MRP-hFg interaction promotes the adherence of S. suis to hCMEC/D3 cells. (A) The adherent ratio of S. suis to hCMEC/D3 cells in the presence of hFg. S. suis 05ZYH33 and 05ZYH33Δmrp mutant were pretreated with a series of concentrations of hFg before infecting a hCMEC/D3 cell monolayer. The adherence ability of S. suis was evaluated by adherence assay. Values represent percent (mean ± S.D.) of total S. suis inoculum bound to the monolayers. (B) The adherent ability of S. suis to hCMEC/D3 cells evaluated by fluorescent microscopy. S. suis 05ZYH33 and 05ZYH33Δmrp mutant were labeled by BCECF (green) before infection, and hCMEC/D3 cell monolayers were treated or untreated with Fg (500 μg/ml). F-actin was strained with Rhodamine-labeled Phalloidin (red), nuclei were stained with DAPI (blue). *P < 0.05.
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Figure 1: MRP-hFg interaction promotes the adherence of S. suis to hCMEC/D3 cells. (A) The adherent ratio of S. suis to hCMEC/D3 cells in the presence of hFg. S. suis 05ZYH33 and 05ZYH33Δmrp mutant were pretreated with a series of concentrations of hFg before infecting a hCMEC/D3 cell monolayer. The adherence ability of S. suis was evaluated by adherence assay. Values represent percent (mean ± S.D.) of total S. suis inoculum bound to the monolayers. (B) The adherent ability of S. suis to hCMEC/D3 cells evaluated by fluorescent microscopy. S. suis 05ZYH33 and 05ZYH33Δmrp mutant were labeled by BCECF (green) before infection, and hCMEC/D3 cell monolayers were treated or untreated with Fg (500 μg/ml). F-actin was strained with Rhodamine-labeled Phalloidin (red), nuclei were stained with DAPI (blue). *P < 0.05.

Mentions: Previous studies have found that S. suis serotype 2 has the ability to adhere to and invade porcine brain microvascular endothelial cells (Vanier et al., 2004). In this study, we evaluated if hFg could affect the adherence of S. suis to human brain microvascular endothelial cells. As shown in Figure 1A, with the increase of hFg, both wild type strain 05ZYH33 and the 05ZYH33Δmrp mutant had increased adherence to hCMEC/D3 cells. The adherent ratio of strain 05ZYH33 was significantly higher than that of the 05ZYH33Δmrp mutant in the presence of hFg. Fluorescent microscopy (Figure 1B) showed that in the presence of hFg, there was significantly more BCECF-labeled S. suis 05ZYH33 than that of the 05ZYH33Δmrp mutant. These results indicate that the interaction of MRP and Fg promotes the adherence of S. suis to hCMEC/D3 cell.


Interaction of fibrinogen and muramidase-released protein promotes the development of Streptococcus suis meningitis.

Wang J, Kong D, Zhang S, Jiang H, Zheng Y, Zang Y, Hao H, Jiang Y - Front Microbiol (2015)

MRP-hFg interaction promotes the adherence of S. suis to hCMEC/D3 cells. (A) The adherent ratio of S. suis to hCMEC/D3 cells in the presence of hFg. S. suis 05ZYH33 and 05ZYH33Δmrp mutant were pretreated with a series of concentrations of hFg before infecting a hCMEC/D3 cell monolayer. The adherence ability of S. suis was evaluated by adherence assay. Values represent percent (mean ± S.D.) of total S. suis inoculum bound to the monolayers. (B) The adherent ability of S. suis to hCMEC/D3 cells evaluated by fluorescent microscopy. S. suis 05ZYH33 and 05ZYH33Δmrp mutant were labeled by BCECF (green) before infection, and hCMEC/D3 cell monolayers were treated or untreated with Fg (500 μg/ml). F-actin was strained with Rhodamine-labeled Phalloidin (red), nuclei were stained with DAPI (blue). *P < 0.05.
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Related In: Results  -  Collection

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Figure 1: MRP-hFg interaction promotes the adherence of S. suis to hCMEC/D3 cells. (A) The adherent ratio of S. suis to hCMEC/D3 cells in the presence of hFg. S. suis 05ZYH33 and 05ZYH33Δmrp mutant were pretreated with a series of concentrations of hFg before infecting a hCMEC/D3 cell monolayer. The adherence ability of S. suis was evaluated by adherence assay. Values represent percent (mean ± S.D.) of total S. suis inoculum bound to the monolayers. (B) The adherent ability of S. suis to hCMEC/D3 cells evaluated by fluorescent microscopy. S. suis 05ZYH33 and 05ZYH33Δmrp mutant were labeled by BCECF (green) before infection, and hCMEC/D3 cell monolayers were treated or untreated with Fg (500 μg/ml). F-actin was strained with Rhodamine-labeled Phalloidin (red), nuclei were stained with DAPI (blue). *P < 0.05.
Mentions: Previous studies have found that S. suis serotype 2 has the ability to adhere to and invade porcine brain microvascular endothelial cells (Vanier et al., 2004). In this study, we evaluated if hFg could affect the adherence of S. suis to human brain microvascular endothelial cells. As shown in Figure 1A, with the increase of hFg, both wild type strain 05ZYH33 and the 05ZYH33Δmrp mutant had increased adherence to hCMEC/D3 cells. The adherent ratio of strain 05ZYH33 was significantly higher than that of the 05ZYH33Δmrp mutant in the presence of hFg. Fluorescent microscopy (Figure 1B) showed that in the presence of hFg, there was significantly more BCECF-labeled S. suis 05ZYH33 than that of the 05ZYH33Δmrp mutant. These results indicate that the interaction of MRP and Fg promotes the adherence of S. suis to hCMEC/D3 cell.

Bottom Line: Muramidase-released protein (MRP) is as an important virulence marker of Streptococcus suis (S. suis) serotype 2.In this study, we found that the deletion of mrp significantly impairs the hFg-mediated adherence and traversal ability of S. suis across human cerebral microvascular endothelial cells (hCMEC/D3).Measurement of the permeability to Lucifer yellow in vitro and Evans blue extravasation in vivo show that the MRP-hFg interaction significantly increases the permeability of the blood-brain barrier (BBB).

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences , Beijing, China ; Urumqi Ethnic Cadres' College , Urumqi, China.

ABSTRACT
Muramidase-released protein (MRP) is as an important virulence marker of Streptococcus suis (S. suis) serotype 2. Our previous works have shown that MRP can bind human fibrinogen (hFg); however, the function of this interaction in S. suis meningitis is not known. In this study, we found that the deletion of mrp significantly impairs the hFg-mediated adherence and traversal ability of S. suis across human cerebral microvascular endothelial cells (hCMEC/D3). Measurement of the permeability to Lucifer yellow in vitro and Evans blue extravasation in vivo show that the MRP-hFg interaction significantly increases the permeability of the blood-brain barrier (BBB). In the mouse meningitis model, wild type S. suis caused higher bacterial loads in the brain and more severe histopathological signs of meningitis than the mrp mutant at day 3 post-infection. Western blot analysis and immunofluorescence observations reveal that the MRP-hFg interaction can destroy the cell adherens junction protein p120-catenin of hCMEC/D3. These results indicate that the MRP-hFg interaction is important in the development of S. suis meningitis.

No MeSH data available.


Related in: MedlinePlus