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Nutritional Immunology and Inflammation/Immunometabolism (Poster Session)

(P14-013-24) The Effects of Dietary Vitamin D Supplementation and In Vitro Vitamin D Treatment on Immunometabolism in Bone Marrow-Derived Dendritic Cells From Control and Diabetic Mice

Sunday, June 30, 2024
12:45 PM – 1:45 PM CT
Location: Poster Board 482

    Presenting Author(s)

  • SJ

    Sohee Jung (she/her/hers)

    graduate student
    Seoul National University
    Seoul, Seoul-t'ukpyolsi, Republic of Korea

    • Co-Author(s)

  • YK

    Yoon-ah Kim

    Seoul National University, Republic of Korea

  • MO

    Minha Oh

    Seoul National University, Republic of Korea

  • GL

    Ga Young Lee, MS

    PhD student
    Seoul National University
    Seoul, Seoul-t'ukpyolsi, Republic of Korea

  • SY

    Soomin Yoo

    Seoul National University, Republic of Korea

  • SH

    Sung Nim Han, PhD, RD

    Professor
    Seoul National University
    Seoul, Seoul-t'ukpyolsi, Republic of Korea

Disclosure(s):
Sohee Jung: No relevant financial relationship(s) with ineligible companies to disclose.


Objectives: The aim of this study is to investigate the effects of dietary vitamin D supplementation and in vitro 1,25(OH)2D3 treatment on differentiation, maturation and metabolism of bone marrow-derived dendritic cells (BMDCs) from diabetic mice.

Methods: Five-week-old C57BLKS/J lar- Leprdb/Leprdb (db/db) and m+/m+ (CON) mice were fed with vitamin D-supplemented diet (10,000 IU/kg diet: vDS) or control diet (1,000 IU/kg diet: vDC) for 8 weeks. BMDCs were generated by culturing bone marrow cells isolated from the mice with rmGM-CSF (20 ng/mL) in the absence or presence of 10 nM 1,25(OH)2D3. The maturation of BMDCs was induced by stimulation with lipopolysaccharides (50 ng/mL) for 24 hours. The expression of surface markers related to immunostimulatory functions (MHC class II, CD80, and CD86) was assessed using flow cytometry analysis. The real-time analysis of oxygen consumption rate (OCR) was conducted using Seahorse XF extracellular flux analyzer to assess mitochondrial function. The mRNA levels of genes related to mitochondrial function and fatty acid metabolism (Prkaa1, Pparg, Cpt1a, and Mpc1) were determined using real-time PCR.

Results: db/db group showed lower surface expression of MHC class II and CD80 than CON group. In vitro 1,25(OH)2D3 treatment decreased the expression of MHC class II, CD80 and CD86 on BMDCs from both CON and db/db groups. In mature BMDCs, db/db group showed higher mitochondrial basal respiration and ATP production levels than CON group. In vitro 1,25(OH)2D3 treatment resulted in decreased mitochondrial maximal respiration and ATP production levels in immature BMDCs from CON and db/db groups and decreased proton leakage in mature BMDCs from CON-vDC, db/db-vDC and CON-vDS groups. In mature BMDCs, mRNA levels of Cpt1a and Mpc1 were higher in db/db group compared to CON group. In vitro treatment of 1,25(OH)2D3 increased Prkaa1 and Cpt1a mRNA levels in CON-vDS and db/db-vDS groups, Pparg mRNA levels in CON-vDS, db/db-vDC, and db/db-vDS groups, and Mpc1 mRNA levels in only CON-vDS group.

Conclusions: These results suggest that vitamin D induce immunotolerance in BMDCs from both control and diabetic mice by affecting mitochondrial function and fatty acid metabolism.

Funding Sources: Supported by the National Research Foundation of Korea (NRF-2021R1A2C2012013)