1. Volume 150, Issue 7, Pages 1620-1632 (June 2016)
Core Fucosylation on T Cells, Required for Activation of T-Cell Receptor Signaling and Induction of Colitis in Mice, Is Increased in Patients With Inflammatory Bowel Disease 
Hironobu Fujii, Shinichiro Shinzaki, Hideki Iijima, Kana Wakamatsu, Chizuru Iwamoto, Tomoaki Sobajima, Ryusuke Kuwahara, Satoshi Hiyama, Yoshito Hayashi, Shinji Takamatsu, Naofumi Uozumi, Yoshihiro Kamada, Masahiko Tsujii, Naoyuki Taniguchi, Tetsuo Takehara, Eiji Miyoshi 
Gastroenterology 
Volume 150, Issue 7, Pages (June 2016)
DOI: /j.gastro
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2. Figure 1
Core fucosylation levels in T cells were increased in colitic conditions. (A) Flow cytometric analysis of cell-surface fucosylation levels of splenocytes in mice treated with TNBS or vehicle control, and (B) mean fluorescence intensity (MFI) (n = 4 per group). (C–F) Flow cytometric analysis of cell-surface fucosylation levels of splenocytes (C) and CD4+ T cells (E) stimulated with anti-CD3ε and anti-CD28 mAbs or vehicle control for 24 hours. (D) MFI related to (C); (F) MFI related to (E) (n = 4 per group). Similar results were obtained in 2 different experiments. ALL, Aleuria aurantia lectin. Data are shown as mean ± SEM. *P < .01.
Gastroenterology  , DOI: ( /j.gastro )
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3. Figure 2
Core fucosylation levels of T cells were increased in inflamed mucosa in IBD patients. (A) FUT8 expression levels in inflamed and noninflamed mucosa from the same patient with Crohn’s disease (n = 8 per group). (B, C) Representative micrographs of immunofluorescence staining of CD3 and PhoSL (B) and the percentage of CD3+ PhoSL+ T cells (C) in inflamed and noninflamed mucosa from same Crohn’s disease patient (n = 4 per group). DAPI, 4′,6-diamidino-2-phenylindole. Scale bar = 30 μm. Data are shown as mean ± SEM. #P < .05.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2016 AGA Institute Terms and Conditions

4. Figure 3
Amelioration of TNBS-induced colitis in Fut8−/− mice. (A) Body weight of Fut8+/+ and Fut8−/− mice subjected to TNBS (n = 6 per group). (B) Micrographs of H&E-stained colon. Scale bar = 200 μm. (C) Histologic scores of TNBS-induced colitis (n = 6 per group). (D) Cytokine production in Fut8+/+ and Fut8−/− mice. MLN isolated from mice subjected to TNBS were cultured with anti-CD3ε and anti-CD28 mAbs for 48 hours, and cytokine levels in the culture supernatants were determined by enzyme-linked immunosorbent assay (n = 6 per group). IFN, interferon. (E) mRNA expression levels in the colon from Fut8+/+ and Fut8−/− mice subjected to TNBS were analyzed by real-time reverse transcription polymerase chain reaction (n = 6 per group). Similar results were obtained in 2 different experiments. Data are shown as mean ± SEM. #P < .05, *P < .01.
Gastroenterology  , DOI: ( /j.gastro )
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5. Figure 4
T cells from Fut8−/− mice exhibited significantly less severe colitis in the adoptive transfer colitis model. (A) Body weights of Rag2−/− mice adoptively transferred with CD4+ CD62L+ naïve T cells from Fut8+/+ or Fut8−/− mice (n = 6 per group). (B, C) Representative micrographs of H&E-stained colon (B) and histological scores (C, n = 6 per group) of adoptively transferred Rag2−/− mice. Scale bar = 200 μm. (D) Cytokine production in adoptively transferred Rag2−/− splenocytes. Isolated splenocytes were incubated with anti-CD3ε and anti-CD28 mAbs for 48 hours and cytokine productions in the culture supernatants were determined by enzyme-linked immunosorbent assay. IFN, interferon. (E) The gene expression levels in the colon from adoptively transferred Rag2−/− mice by real-time reverse transcription polymerase chain reaction (n = 6 per group). Similar results were obtained in 2 different experiments. Data are shown as mean ± SEM. #P < .05; *P < .01.
Gastroenterology  , DOI: ( /j.gastro )
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6. Figure 5
Decreased production of both Th1 and Th2 cytokines in CD4+ T cells from Fut8−/− mice. Purified splenic CD4+ T cells from Fut8+/+ and Fut8−/− mice were cultured with anti-CD3ε and anti-CD28 mAbs, and cytokine productions were analyzed. Gene expression levels were measured by real-time RT-PCR (A) and cytokine levels were measured by ELISA (B) (n = 4 per group). Similar results were obtained in 2 different experiments. IFN, interferon. Data are shown as mean ± SEM. *P < .01.
Gastroenterology  , DOI: ( /j.gastro )
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7. Figure 6
Suppression of T-cell signaling through impaired transport of TCR to lipid rafts in CD4+ T cells of Fut8−/− mice. (A) Cell proliferation of CD4+ T cells from Fut8+/+ and Fut8−/− incubated with anti-CD3ε and anti-CD28 mAbs for 48 hours (n = 4 per group). Data are shown as mean ± SEM. *P < .01 (B) Western blot (WB) analysis of CD4+ T cells from Fut8+/+ and Fut8−/− mice incubated with anti-CD3ε and anti-CD28 mAbs. (C) Immunoprecipitation (IP) with anti-TCRβ and anti-CD28 antibodies, followed by lectin blot analysis using Aspergillus oryzae lectin (AOL). (D) CD4+ T cells from Fut8+/+ and Fut8−/− mice were incubated with anti-CD3ε and anti-CD28 mAbs for 10 minutes, and expression levels of TCRβ, CD28, CD3ε, and flotillin-1 in the lipid raft fraction were analyzed by WB analysis. (E) Immunofluorescence staining of TCRβ (red) and flotillin-1 (green) in splenocytes from Fut8+/+ and Fut8−/− mice were incubated with anti-CD3ε and anti-CD28 mAbs for 10 minutes. Scale bar = 50 μm. (F) Immunogold electron microscopy of TCRβ (arrow) and flotillin-1 (arrowhead) in splenocytes from Fut8+/+ and Fut8−/− mice were incubated with anti-CD3ε and anti-CD28 mAbs for 10 minutes. Scale bar = 500 nm. Similar results were obtained in 3 different experiments.
Gastroenterology  , DOI: ( /j.gastro )
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