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Link to original content: http://www.ncbi.nlm.nih.gov/pubmed/31272068
Vitamin D/Vitamin D Receptor Signaling Is Required for Normal Development and Function of Group 3 Innate Lymphoid Cells in the Gut - PubMed Skip to main page content
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. 2019 Jul 26:17:119-131.
doi: 10.1016/j.isci.2019.06.026. Epub 2019 Jun 20.

Vitamin D/Vitamin D Receptor Signaling Is Required for Normal Development and Function of Group 3 Innate Lymphoid Cells in the Gut

Lei He  1 , Min Zhou  2 , Yan Chun Li  3
Affiliations

Vitamin D/Vitamin D Receptor Signaling Is Required for Normal Development and Function of Group 3 Innate Lymphoid Cells in the Gut

Lei He et al. iScience. .

Abstract

Group 3 innate lymphoid cells (ILC3) play key roles in protective immunity and mucosal barrier maintenance. Here we showed that vitamin D/vitamin D receptor (VDR) signaling regulates gut ILC3. VDR deletion or 1,25-dihydroxyvitamin D deficiency in mice led to a marked reduction in colonic ILC3 populations at steady state and impaired ILC3 responses following Citrobacter rodentium infection, resulting in substantial increases in intestinal bacterial growth and mouse mortality. VDR regulation of ILC3 was independent of T and B lymphocytes or gut microflora. Correction of 1,25-dihydroxyvitamin D deficiency rescued the ILC3 defects. Mechanistically, VDR deletion or 1,25-dihydroxyvitamin D deficiency markedly reduced colonic Ki67+ ILC3 populations, and in vivo and in vitro studies confirmed that vitamin D hormone directly stimulated ILC3 proliferation. Therefore, vitamin D/VDR signaling is required for ILC3-mediated innate immunity through regulation of ILC3 proliferation.

Keywords: Biomolecules; Components of the Immune System; Immune Response; Immunology.

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Conflict of interest statement

The authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
Global VDR Deletion Impairs Gut ILC3 Development in Mice Colonic lamina propria (LP) cells were isolated from WT and VDR−/− mice, and the cells were analyzed by FACS for ILC3 populations. (A–C) Representative FACS plots for analysis of RORγt+ILC3 (A), NKp46+ cells (B), and LTi4 and LTi0 cells (C) in WT and VDR−/− mice at steady state. (D–F) Quantitation based on FACS data of RORγt+ILC3 (D), NKp46+ cells (E), and LTi4 and LTi0 cells (F) in WT and VDR−/− mice. The data were presented as percentage of the gated population and absolute cell number. **p < 0.01; ***p < 0.001. n = 4 each genotype. Data are represented as mean ± SEM.
Figure 2
Figure 2
Global VDR Deletion Leads to Impaired Gut Immunity against C. rodentium WT and VDR−/− mice were gavaged with C. rodentium. The mice were monitored daily for 16 days post infection. The mice were killed on day 5 following infection for FACS analyses of colonic LP cells. (A–C) (A) Body weight changes (n = 10), (B) survival curve (n = 10–11), and (C) fecal daily bacterial counts (CFU) per gram of feces in WT and VDR−/− mice under C. rodentium infection (n = 6–7). p < 0.001 by log rank test. (D–G) Representative FACS plots for the analysis of RORγt+ILC3 (D), IL-22+ILC3 (E), NKp46+ (F), and LTi4 and LTi0 cells (G) in WT and VDR−/− mice at steady state (Ctrl) and under C. rodentium infection. (H–J) Quantitation RORγt+ILC3 and IL-22+ILC3 (H), NKp46+ cells (I), and LTi4 and LTi0 cells (J) in WT and VDR−/− mice at baseline and under C. rodentium infection. The data were presented as percentage of the gated population and absolute cell number. *p < 0.05; **p < 0.01; ***p < 0.001. n = 4 each group. Data are represented as mean ± SEM.
Figure 3
Figure 3
Deficiency in 1,25(OH)2D3 Synthesis Impairs ILC3 Development and Its Immunity against C. rodentium WT and Cyp27b1−/− mice were gavaged with C. rodentium. The mice were killed on day 5 following infection for FACS analyses of colonic LP cells. (A and B) (A) Body weight changes; (B) fecal bacterial counts per gram of feces in WT and Cyp27b1−/− mice on days 2 and 5 following C. rodentium infection. ***p < 0.001 vs. WT. n = 4–5 each group. (C–F) Representative FACS plots for the analysis of RORγt+ILC3 (C), IL-22+ILC3 (D), NKp46+ (E), and LTi4 and LTi0 cells (F) in WT and Cyp27b1−/− mice at steady state (Ctrl) and under C. rodentium infection. (G–I) Quantitation RORγt+ILC3 and IL-22+ILC3 (G), NKp46+ cells (H), and LTi4 and LTi0 cells (I) in WT and Cyp27b1−/− mice at baseline and under C. rodentium infection. The data were presented as percentage of the gated population and absolute cell number. *p < 0.05; **p < 0.01; ***p < 0.001. n = 4 each group. Data are represented as mean ± SEM.
Figure 4
Figure 4
ILC3-specific Deletion of VDR Impairs ILC3 Development and Its Immunity against C. rodentium Infection VDRflox/flox and VDRflox/flox;RORγt-Cre mice were gavaged with C. rodentium. The mice were killed on day 4 following infection. (A and B) (A) Body weight changes; (B) fecal bacterial counts per gram of feces in VDRflox/flox and VDRflox/flox;RORγt-Cre mice on day 4 after C. rodentium infection. ***p < 0.001 vs. VDRflox/flox. n = 4–5 each genotype. (C–F) Representative FACS plots for the analysis of RORγt+ILC3 (C), IL-22+ILC3 (D), NKp46+ (E), and LTi4 and LTi0 cells (F) in VDRflox/flox and VDRflox/flox;RORγt-Cre mice at steady state (Ctrl) and under C. rodentium infection. (G–I) Quantitation RORγt+ILC3 and IL-22+ILC3 (G), NKp46+ cells (H), and LTi4 and LTi0 cells (I) in VDRflox/flox and VDRflox/flox;RORγt-Cre mice at baseline and under C. rodentium infection. The data were presented as percentage of the gated population and absolute cell number. *p < 0.05; **p < 0.01; ***p < 0.001. n = 4 each group. Data are represented as mean ± SEM.
Figure 5
Figure 5
Bone Marrow (BM) Transplantation Confirms VDR Deletion in ILC3 Leading to Impaired ILC3 Development BM cells obtained from VDRflox/flox or VDRflox/flox/RORγt-Cre mice were transplanted to recipient Rag1−/− mice 6 h after receiving lethal γ-irradiation. The recipient mice were analyzed 8 weeks after transplantation. (A and B) (A) Representative FACS plots for RORγt+ILC3 analyses; (B) quantitation of colonic RORγt+ILC3, NKp46+, and LTi4 and LTi0 cells in recipient mice transplanted with VDRflox/flox or VDRflox/flox;RORγt-Cre BM cells. The data were presented as percentage of the gated population and absolute cell number. *p < 0.05, **p < 0.01. n = 3–4 each group. Data are represented as mean ± SEM.
Figure 6
Figure 6
Deletion of VDR or Cyp27b1 Reduces ILC3 Proliferation Colonic LP cells were isolated from WT or VDR−/− mice at steady state (Ctrl) or under C. rodentium infection for 5 days (A and B), from VDRflox/flox or VDRflox/flox;RORγt-Cre mice at steady state or under C. rodentium infection for 5 days (Cand D), or from WT and Cyp27b1−/− mice at steady state (E and F). Ki67+ ILC3 and ILC3 subsets were quantified by FACS. (A, C, and E) Representative FACS plots for Ki67+ RORγt+ILC3; (B, D, and F) quantitation of Ki67+RORγt+ILC3 and Ki67+NKp46+, Ki67+LTi4, and Ki67+LTi0 subsets. *p < 0.05; **p < 0.01; ***p < 0.001. n = 4–5 each group. Data are represented as mean ± SEM.
Figure 7
Figure 7
Ligand Activation of VDR Stimulates ILC3 Proliferation (A) FACS quantitation of colonic Ki67+RORγt+ILC3 and Ki67+NKp46+, Ki67+LTi4, and Ki67+LTi0 subpopulations in WT, Cyp27b1−/− or Cyp27b1−/− mice treated with 1,25(OH)2D3 for 1 week; (B) FACS quantitation of colonic Ki67+RORγt+ILC3 and Ki67+NKp46+, Ki67+LTi4, and Ki67+LTi0 subsets in WT untreated or treated with paricalcitol for 1 week; (C) Lin−Thy1.2hiKLRG1−IL-7Rα+ cells were sorted by FACS and cultured in the presence or absence of 1,25(OH)2D3 for 3 days, and Ki67+RORγt+ILC3 population was analyzed by FACS. Shown are representative FACS plots. *p < 0.05, **p < 0.01; ***p < 0.001. n = 4 each group. Data are represented as mean ± SEM.
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