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  • Ascorbic acid modulates cell migration in differentiated HL-60 cells and peripheral blood leukocytes.

Ascorbic acid modulates cell migration in differentiated HL-60 cells and peripheral blood leukocytes.

Molecular nutrition & food research (2015-03-27)
Joseph Schwager, Albine Bompard, Peter Weber, Daniel Raederstorff
ABSTRACT

The impact of L-ascorbic acid (L-AA) on the chemokinesis (CK) and chemotaxis (CT) of HL-60 cells and polymorphonuclear cells (PMN) was investigated. HL-60 cells were differentiated with DMSO, retinoic acid (RA), vitamin D, or L-AA. Chemokinesis and chemotaxis of differentiated HL-cells were assayed. Vitamin D3-treated HL-60 cells (dHL-60vitD3 cells) and RA-treated cells (dHL-60RA cells) acquired monocyte/macrophage-like and neutrophil-like phenotypes, respectively. DMSO induced the differentiation of an intermediate phenotype (dHL-60DMSO cells), whereas L-AA downregulated neutrophil markers (dHL-60L-AA cells). dHL-60DMSO cells had increased CK and potent CT in gradients of IL-8 and N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP). dHL-60RA cells and dHL-60L-AA cells migrated less toward IL-8 and fMLP; dHL-60vitD3 cells preferably responded to fMLP. L-AA enhanced CK of dHL-60DMSO cells and was a weak chemo-attractant. In human leukocytes, IL-8 and fMLP triggered receptor-mediated chemotaxis. CXCR2 and fMLPR were downregulated by IL-8 and fMLP, respectively. L-AA stimulated chemotaxis although significantly less than IL-8 and fMLP. IL-8 targeted chemotaxis was enhanced both in HL-60 cells and leukocytes when cells were incubated with L-AA. L-AA modulated chemokinesis and had significant chemo-attractant properties, which were independent on fMLP or IL-8 receptors. The results suggest that L-AA improves leukocyte function in innate immune responses.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
25-Hydroxyvitamin D3 solution, 5 μg/mL in ethanol, 98% (CP)
Sigma-Aldrich
25-Hydroxyvitamin D3 solution, 50 μg/mL in ethanol, 98% (CP)
Sigma-Aldrich
25-Hydroxyvitamin D3 solution, 100 μg/mL in ethanol, 98% (CP)
Sigma-Aldrich
25-Hydroxyvitamin D3 monohydrate, ≥99.0% (HPLC)
Sigma-Aldrich
Retinoic acid, ≥98% (HPLC), powder
Sigma-Aldrich
25-Hydroxycholecalciferol, ≥98% (HPLC)
Supelco
25-Hydroxyvitamin D3 solution, 100 μg/mL in ethanol, ampule of 1 mL, certified reference material, Cerilliant®