Immune Netw.  2014 Apr;14(2):100-106. 10.4110/in.2014.14.2.100.

Shigella flexneri Inhibits Intestinal Inflammation by Modulation of Host Sphingosine-1-Phosphate in Mice

• Kim YI
• Yang JY
• Ko HJ
• Kweon MN
• Chang SY

• KMID: 2168020
• DOI: http://doi.org/10.4110/in.2014.14.2.100

Abstract

Infection with invasive Shigella species results in intestinal inflammation in humans but no symptoms in adult mice. To investigate why adult mice are resistant to invasive shigellae, 6~8-week-old mice were infected orally with S. flexneri 5a. Shigellae successfully colonized the small and large intestines. Mild cell death was seen but no inflammation. The infected bacteria were cleared 24 hours later. Microarray analysis of infected intestinal tissue showed that several genes that are involved with the sphingosine-1-phosphate (S1P) signaling pathway, a lipid mediator which mediates immune responses, were altered significantly. Shigella infection of a human intestinal cell line modulated host S1P-related genes to reduce S1P levels. In addition, co-administration of S1P with shigellae could induce inflammatory responses in the gut. Here we propose that Shigella species have evasion mechanisms that dampen host inflammatory responses by lowering host S1P levels in the gut of adult mice.

Keyword

Shigella flexneri; Inflammation; Sphingosine-1-phosphate; Intestine

MeSH Terms

Adult
Animals
Bacteria
Cell Death
Cell Line
Colon
Humans
Inflammation*
Intestines
Mice*
Microarray Analysis
Shigella
Shigella flexneri*

Figure

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  Figure 1 S. flexneri evade and evoke host cell death in the small intestine without inflammation. (A) Hematoxylin and eosin (H&E) staining of terminal ileum 1 and 24 h after oral infection with 5×109 CFU wild-type M90T or T3SS mutant M90TΔipaB4 (n=5 per group). (B) Periodic acid-Schiff (PAS) staining of terminal ileum 1 and 3 hours after oral infection with 5×109 CFU M90T. Arrows indicate secretory granules of crypt Paneth cells and mucin of goblet cells (n=3 per group). (C) Localization of GFP expressing M90T in the villi and crypts of the terminal ileum 1 h after oral infection (n=3 per group).

• 

  Figure 2 S. flexneri evade colon epithelium without inflammation. (A) H&E staining of ascending and descending colon orally infected with M90T after 3 and 24 h (n=5 per group). (B) M90T-GFP in the epithelium and lamina propria of the colon 3 h after oral infection (n=3 per group). (C) CFU of M90T in the colon and cecum 1, 3, and 24 h after infection. Graphs show mean±SEM. (one-way ANOVA with Tukey post test, ***p<0.0001).

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  Figure 3 S. flexneri modulate host gene expression. (A) Microarray analysis of ileal tissue uninfected or infected with M90T (1 or 16 h after infection) and T3SS mutant M90TΔipaB4 (1 h). (B) Representative gene expression of ileal tissue infected with M90T (1 h after infection) regarding lipid metabolism. (C) mRNA expression of Spgl1 and Ppap2a of ileal tissue 1 h after oral infection with M90T or wild-type Salmonella typhimurium (UK-1).

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  Figure 4 S. flexneri modulate host enzymes to regulate S1P level. (A) CFUs of avirulent BS176 and M90T in Caco-2 cells 2 h following infection. ***p<0.0001. (B) S1P is formed by phosphorylation of sphingosine that is catalyzed by sphingosine kinase (SK) and breaks down by S1P lyase (S1PL) and S1P phosphatase (S1PP). (C) mRNA expression of S1PL, S1PP1, S1PP2, SK1, and SK2 of Caco-2 cells infected with avirulent BS176 (3 h) or M90T (1 and 3 h).

• 

  Figure 5 S. flexneri-provoked inflammation in the gut of adult mice in the presence of S1P. (A) Terminal ileum was looped and M90T injected directly into the looped tissue with or without S1P. After 1 h and 24 h, H&E staining was performed. Representative data were selected from three repetitive experiment sets. (B and C) Pathological grade of the looped ileum tissue injected with M90T and S1P after 1 h (B) and 24 h (C). **p<0.001, ***p<0.0001 (one-way ANOVA with Tukey post test).

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