Immune Netw.  2015 Jun;15(3):135-141. 10.4110/in.2015.15.3.135.

Alloferon Alleviates Dextran Sulfate Sodium-induced Colitis

• Kim H
• Im JP
• Kim JS
• Kang JS
• Lee WJ

• KMID: 2168039
• DOI: http://doi.org/10.4110/in.2015.15.3.135

Abstract

Dysfunction of gut immune regulation is involved in mucosal damage in inflammatory bowel disease (IBD). However, there is still no efficacious immune-regulator for the treatment of IBD. Alloferon is a novel immune-modulatory peptide that was originally isolated from infected insects. It shows anti-inflammatory effects by the regulation of cytokine production by immune cells and their activities. Therefore, we investigated the effect of alloferon in a mouse model of colitis using dextran sulfate sodium (DSS). Colitis was induced by administration of DSS in drinking water for 7 consecutive days. It was confirmed by the presence of weight loss, diarrhea, hematochezia, and colon contraction. Alloferon was injected 4 days after DSS administration. We found that alloferon improved the pathogenesis of IBD based on the reduced disease activity index (DAI) and colon contraction. Edema, epithelial erosion, and immune cell infiltration were found in mice administered DSS, but the phenomena were reduced following alloferon treatment. The plasma level of IL-6, a classical pro-inflammatory cytokine in colitis, was also decreased by alloferon. Moreover, alloferon inhibited the TNF-alpha-induced degradation and phosphorylation of IkappaB in Colo205 colon cancer cells. Taken together, these results show that alloferon has anti-inflammatory effects and attenuates DSS-induced colitis.

Keyword

Alloferon; Inflammatory bowel disease (IBD); Anti-inflammation; DSS-induced colitis

MeSH Terms

Animals
Colitis*
Colon
Colonic Neoplasms
Dextran Sulfate*
Diarrhea
Drinking Water
Edema
Gastrointestinal Hemorrhage
Inflammatory Bowel Diseases
Insects
Interleukin-6
Mice
Phosphorylation
Plasma
Weight Loss
Dextran Sulfate
Drinking Water
Interleukin-6

Figure

• 

  Figure 1 Experimental design of dextran sulfate sodium (DSS) and alloferon treatment. Mice were treated with 3% DSS administered in drinking water for 7 days. Alloferon (50 µg/mouse, i.p.) was daily injected to mice four times from day 4 of DSS treatment.

• 

  Figure 2 Alloferon decreased the severity of dextran sulfate sodium (DSS)-induced colitis. (A) Body weight was monitored for 7 days following 3% DSS and alloferon (50 µg/mouse) treatment. (B) After DSS and alloferon treatment for 7 days, colon length of each experimental group was measured (n=three, *p<0.05, **p<0.01). (C) Body weight loss, stool consistency, and bloody stool were presented as DAI according to the criteria described in the Materials and Methods. DAI calculated as, (stool consistency+gross bleeding+weight loss)/3, *p<0.05, ***p<0.001. DAI, disease activity index; ctl, control; Allo, alloferon.

• 

  Figure 3 Alloferon decreased inflammation and IL-6 production in dextran sulfate sodium (DSS)-induced colitis. (A) Colons were longitudinally sectioned and stained with H&E. Scale bar, 200 and 100 µm (upper and lower panels, respectively). (B) Cross-sectioned colons were stained with H&E and severity was scored according to criteria described in Materials and Methods (n=three, **p<0.01. (C) After DSS and alloferon treatment, the blood was collected in heparinized capillaries. After centrifugation, the plasma concentration of IL-6 was measured using ELISA, *p<0.05; ctl, control; Allo, alloferon.

• 

  Figure 4 Alloferon inhibited Iκ B degradation. Colo205 cells (1×106) were treated with TNF-α (10 ng/ml) for 10 min after alloferon pre-treatment (1 µg) for 24 h. The expression of IκB and phospho-(p)-IκB was determined using immunoblotting. β-actin was used as a loading control.

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