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14.09.2017 | Original Contribution | Ausgabe 7/2018

European Journal of Nutrition 7/2018

High-dose vitamin D3 supplementation decreases the number of colonic CD103+ dendritic cells in healthy subjects

Zeitschrift:
European Journal of Nutrition > Ausgabe 7/2018
Autoren:
Nina Friis Bak, M. Bendix, S. Hald, L. Reinert, M. K. Magnusson, J. Agnholt
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1007/​s00394-017-1531-y) contains supplementary material, which is available to authorized users.

Abstract

Purpose

Vitamin D may induce tolerance in the intestinal immune system and has been shown to regulate the phenotype of tolerogenic intestinal dendritic cells (DCs) in vitro. It is unknown whether vitamin D supplementation affects human intestinal DCs in vivo, and we aimed to investigate the tolerability and effect on intestinal CD103+DCs of high-dose vitamin D3 treatment in healthy subjects.

Methods

Ten healthy subjects received a total of 480,000 IU oral vitamin D3 over 15 days and colonic biopsies were obtained before and after intervention by endoscopy. Lamina propria mononuclear cells (LPMCs) were isolated from the biopsies, stained with DC surface markers and analysed with flow cytometry. Snap-frozen biopsies were analysed with qPCR for DC and regulatory T cell-related genes.

Results

No hypercalcemia or other adverse events occurred in the test subjects. Vitamin D decreased the number of CD103+ DCs among LPMCs (p = 0.006). Furthermore, vitamin D induced mRNA expression of TGF-β (p = 0.048), TNF-α (p = 0.006) and PD-L1 (p = 0.02) and tended to induce IL-10 expression (p = 0.06). Multivariate factor analysis discriminated between pre- and post-vitamin D supplementation with a combined increased qPCR expression of PD1, PD-L1, TGF-β, IL-10, CD80, CD86, FOXP3, NFATc2 and cathelicidin.

Conclusion

High-dose vitamin D supplementation is well tolerated by healthy subjects and has a direct effect on the CD103+ DCs, local cytokine and surface marker mRNA expression in the colonic mucosa, suggestive of a shift towards a more tolerogenic milieu.

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Zusatzmaterial
Supplementary figure S1 DC’s (HLA-DR+, CD11c+, CD14−, blue colour) backgated to the live cells gate, defined as 7AAD− (TIFF 4448 kb)
394_2017_1531_MOESM1_ESM.tiff
Supplementary Table S1 List of TaqMan Gene Expression Assays used in the experiment. All products from Applied Biosystems, Life Technologies, Carlsbad, California, USA. The assay ID provides information on NCBI reference sequence number, amplicon length and location of each primer by exon or intron. All TaqMan Gene Expression Assays have amplification efficiencies of 100% ± 10% and will amplify the intended target at least 10 Cq earlier than the gene with the closest sequence homology. Supplementary Table S2 List of genes analysed by qPCR. Medians are listed for baseline normalised Cq-values and day 15 normalised Cq-values. IFNG is excluded because of too many missing values. P-value is given for paired non-parametric analyses (Wilcoxon matched-pairs signed-rank test) (PDF 84 kb)
394_2017_1531_MOESM2_ESM.pdf
Literatur
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