Intest Res.  2020 Oct;18(4):402-411. 10.5217/ir.2019.00132.

A glycolipid adjuvant, 7DW8-5, provides a protective effect against colonic inflammation in mice by the recruitment of CD1d-restricted natural killer T cells

• Lee C
• Hong SN
• Kim YH

• KMID: 2508566
• DOI: http://doi.org/10.5217/ir.2019.00132

Abstract

Background/Aims
The modulation of CD1d-restricted natural killer T (NKT) cells by glycolipids has been considered as a potential therapy against immunologic diseases, including inflammatory bowel disease. A recently identified a glycolipid analog, 7DW8-5, which is derived from α-galactosylceramide (α-GalCer), is as much as 100-fold more active at stimulating both human and mice NKT cells when compared to α-GalCer. We explored the effects of 7DW8-5 in mouse models of acute and chronic colitis.
Methods
We investigated the effects of 7DW8-5 on intestinal inflammation by assessing the effects of 7dW8-5 on a murine dextran sulfate sodium (DSS)-induced acute colitis model and a chronic colitis-associated tumor model.
Results
The acute DSS-induced colitis model showed a dose-dependent response to 7DW8-5, as mice administered 7DW8-5 showed a significant improvement in DSS-induced colitis based on their disease activity index, histologic analysis, and serum C-reactive protein levels, when compared to mice administered vehicle alone. However, DSS-induced colitis in CD1d-KO mice showed no response to 7DW8-5. A fluorescence-activating cell sorting analysis revealed an increase in NKT cells in colonic tissues of 7DW8-5-treated mice. RNA-seq and real-time quantitative polymerase chain reaction showed a significant increase in the expression of interleukin (IL)-4, IL-13, and interferon-gamma in 7DW8-5-treated mice. In addition, 7DW8-5 treatment reduced colitis-associated tumor development in an azoxymethane/DSS mouse model.
Conclusions
7DW8-5 activates NKT cells through CD1d and provides a protective effect against intestinal inflammation in mice. Therefore, 7DW8-5 may be a promising therapeutic agent for treatment of inflammatory bowel disease.

Keyword

7DW8-5; Alpha-galactosylceramide; Natural killer T-cells; Dextran sulfate sodium; Inflammatory bowel disease

Figure

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  Fig. 1. Structure of α-galactosylceramide (α-GalCer) and 7DW8-5.

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  Fig. 2. The effect of 7DW8-5 on dextran sulfate sodium (DSS)-induced acute colitis. (A) The changes in body weight. (B) Disease activity index. (C) Histological score. Magnified image of the top black square is the bottom picture (H&E). (D) Serum C-reactive protein level. (E) Colon length. aP<0.05, bP<0.01, cP<0.001. α-GalCer, α-galactosylceramide.

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  Fig. 3. The effect of 7DW8-5 on dextran sulfate sodium (DSS)-induced acute colitis in CD1d-/- mice and anti-NK1.1 administrated C57BL/6 mice. (A) The changes in body weight. (B) Colon length. (C) Disease activity index. (D) Histological score.

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  Fig. 4. Fluorescence-activating cell sorting for invariant natural killer T (iNKT) cell. (A) Gating strategy for flow cytometry analysis. (B) Percentage of NKT cell population in the colon tissue of the DSS/7DW8-5 group. FSC, forward scatter; DSS, dextran sulfate sodium.

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  Fig. 5. The effect of 7DW8-5 in the chronic colitis-induced tumor model. (A) Colon length. (B) The number of tumors ≥1 mm. AOM, azoxymethane; DSS, dextran sulfate sodium.

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  Fig. 6. Expression pattern of T helper type 1 (Th1) and Th2 cytokines in 7DW8-5 or vehicle only administrated dextran sulfate sodium (DSS)-induced acute colitis model. (A) Heatmap of differentially expressed cytokine genes. (B) Real-time quantitative polymerase chain reaction analysis for interleukin (IL)-4, IL-13, and interferon γ (IFN-γ).

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