Cd1d-dependent regulation of bacterial colonization in the intestine of mice
Edward E.S. Nieuwenhuis, … , Andrew B. Onderdonk, Richard S. Blumberg
Edward E.S. Nieuwenhuis, … , Andrew B. Onderdonk, Richard S. Blumberg
Published April 6, 2009
Citation Information: J Clin Invest. 2009;119(5):1241-1250. https://doi.org/10.1172/JCI36509.
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Research Article Gastroenterology

Cd1d-dependent regulation of bacterial colonization in the intestine of mice

Abstract

The accumulation of certain species of bacteria in the intestine is involved in both tissue homeostasis and immune-mediated pathologies. The host mechanisms involved in controlling intestinal colonization with commensal bacteria are poorly understood. We observed that under specific pathogen–free or germ-free conditions, intragastric administration of Pseudomonas aeruginosa, E. coli, Staphylococcus aureus, or Lactobacillus gasseri resulted in increased colonization of the small intestine and bacterial translocation in mice lacking Cd1d, an MHC class I–like molecule, compared with WT mice. In contrast, activation of Cd1d-restricted T cells (NKT cells) with α-galactosylceramide caused diminished intestinal colonization with the same bacterial strains. We also found prominent differences in the composition of intestinal microbiota, including increased adherent bacteria, in Cd1d–/– mice in comparison to WT mice under specific pathogen–free conditions. Germ-free Cd1d–/– mice exhibited a defect in Paneth cell granule ultrastructure and ability to degranulate after bacterial colonization. In vitro, NKT cells were shown to induce the release of lysozyme from intestinal crypts. Together, these data support a role for Cd1d in regulating intestinal colonization through mechanisms that include the control of Paneth cell function.

Authors

Edward E.S. Nieuwenhuis, Tetsuya Matsumoto, Dicky Lindenbergh, Rob Willemsen, Arthur Kaser, Ytje Simons-Oosterhuis, Sylvia Brugman, Keizo Yamaguchi, Hiroki Ishikawa, Yuji Aiba, Yasuhiro Koga, Janneke N. Samsom, Kenshiro Oshima, Mami Kikuchi, Johanna C. Escher, Masahira Hattori, Andrew B. Onderdonk, Richard S. Blumberg

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Figure 6

Morphological and functional differences of Pc granules in Cd1d–/– mice and upon NKT cell activation.

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Morphological and functional differences of Pc granules in Cd1d–/– mice ...
(A) Decreased lysozyme-p immunoreactivity in jejuno-ileal sections of WT mice (right panels) at t = 96 hours (bottom panel) of colonization with E. coli compared with baseline levels (top panel), which is absent in Cd1d–/– animals (left panels). Germ-free Cd1d–/– mice exhibited normal Pc degranulation relative to WT animals at 30 minutes after in vivo treatment with pilocarpine (2.0 mg/mouse). (B) Coincubation of crypt preparations with αGalCer and DN32 cells results in lysozyme-p release, expressed as a ratio of the optical intensity for lysozyme-p/optical intensity for β-actin in the pellet × 100. (C) Mice exhibit empty granules, with enhanced lysozyme immunoreactivity along the crypt axis at 24 hours of intraperitoneal injection with 2 μg αGalCer, which are absent in PBS-treated mice. (D) Compared to WT mice, Pcs of Cd1d–/– mice exhibit smaller granules. (E) WT Pc granules have an electron-dense inner domain and a halo of intermediate density (left panel, arrow). In contrast, in Cd1d–/– mice, the peripheral halo is devoid of electron-dense material (right panel, arrow). (F) Histochemical staining with HRP-labeled lectins (Helix pomatia, top panel; Triticum vulgaris, bottom panel) reveals an altered oligosaccharide composition in Pc granules in Cd1d–/– mice (left panels). Original magnification, ×20 (A); ×40 (C); ×9,000 (E); ×60 (F). Data represent mean ± SEM (B and D); *P < 0.05, **P < 0.01.