Data on myeloperoxidase-oxidized low-density lipoproteins stimulation of cells to induce release of resolvin-D1

This article present data related to the publication entitled “Native and myeloperoxidase-oxidized low-density lipoproteins act in synergy to induce release of resolvin-D1 from endothelial cells” (Dufour et al., 2018). The supporting materials include results obtained by Mox-LDLs stimulated macrophages and investigation performed on scavenger receptors. Linear regressions (RvD1 vs age of mice and RvD1 vs CL-Tyr/Tyr) and Data related to validation were also presented. The interpretation of these data and further extensive insights can be found in Dufour et al. (2018) [1].

were also presented. The interpretation of these data and further extensive insights can be found in Dufour et al. (2018) [1].
& 2018 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Subject area Biochemistry
More specific subject area Resolvin D1 in atherosclerosis Type of data Table, text file, graph, figure How data was acquired Mass spectrometry (LC-MS-/-MS system from Agilent Technologies (Santa Clara, CA, USA): an Agilent 1290 Infinity Binary -UHPLC system coupled to a mass spectrometer Agilent Jet Stream electrospray ionization source (AJS)-Triple Quadrupole (QQQ) 6490 series) Rt-PCR Data format Analyzed Experimental factors Samples were treated by liquid-liquid extraction before analysis Experimental features RvD1 and precursors (17S-HDHA and DHA) were quantified in cell supernatant or in plasma. Data source location Brussels, Belgium Data accessibility The data are provided with this article

Value of the data
Validation of method was an important part of this work and we showed how it was developed. Data show how scavenger receptors, usually involved in oxidized LDLs recognition, were analyzed and how they could be involved in RvD1 synthesis.
Correlation was established between level of RvD1 and Cl-Tyr/Tyr ratio from healthy donors and between level of RvD1 and age in mice. These results are illustrated here.
Because HMEC have shown ability to produce RvD1, we assay the production of RvD1 by monocytes (THP-1).

Data
Different aspects of method validation for RvD1 and precursors analysis were described. Moreover, we investigated many aspects of RvD1 mechanistic of production linked to Mox-LDLs stimulation of endothelial cells and THP-1 macrophages.

Macrophages are apparently not essential for RvD1 production
It is generally accepted that EC and monocytes/macrophages are both required to produce RvD1. However, it was shown that EC are able to produce RvD1 alone [1]. Therefore, we tested whether Mox-LDLs induced RvD1 production in the presence of THP-1. Cells were incubated with 100 μg/ml of Mox-LDLs and data showed an increased concentration of RvD1 (76 7 21 pg/ml) compared with THP-1 cells incubated with 100 μg/ml non-physiologic Ox-LDLs (RvD1 35 7 16 pg/ml) or 100 μg/ml LDLs-nat (RvD1 14 7 10 pg/ml). However, the differences were not statistically significant. An increase in 17S-HDHA was also observed in the same conditions with a concentration of 439 7 32 pg/ ml when incubated with 100 μg/ml of Mox-LDLs and concentrations of 192 7 84 pg/ml and 202 7 25 pg/ml when incubated with Ox-LDLs or LDLs-nat, respectively. These data were again not statistically significant (see Fig. 1).

Macrophages are apparently not essential for RvD1 production
Because we suspected that THP-1 may participate in RvD1 production, we also tested the effects of Mox-LDLs, non-physiologic Ox-LDLs and LDLs-nat on the synthesis of RvD1 and 17S-HDHA by THP-1.     2.2. in vivo macrophage subpopulations analysis using flow cytometry 100 µL of total blood were incubated for 15 min at RT with PE mouse anti-human CD14 and V500 mouse anti-human CD16 antibodies (Becton Dickinson, Franklin Lakes, NJ, USA) as well as with mouse anti-human CD86-FITC and anti-human CCR2-APC monoclonal antibodies (Miltenyi Biotec, Bergisch Gladbach, Germany) for determining the M1 polarization or with mouse anti-human CD206-FITC, anti-human CXCR3-APC and anti-human CD163-VioBlue monoclonal antibodies (Miltenyi Biotec, Bergisch Gladbach, Germany) for the M2 polarization. Red blood cells were then eliminated by adding BD FACS Lysing Solution (dilution: 1/20) (Becton Dickinson, Franklin Lakes, NJ, USA) to total blood and remaining cells were washed twice with 1 mL of running buffer. Cells were finally resuspended in 300 µL of running buffer for analysis. The matching isotype controls were used for each antibody in order to define the threshold. The analysis was performed using the MACSQuant Analyzer 10 (Miltenyi Biotec, Bergisch Gladbach, Germany), applying a gating strategy based on the SSC vs PE gate (CD14), selecting the monocyte population. Classical monocytes were defined based on a high expression of   Table 2 Data of validation: Recovery and coefficients of variation (CV) obtained using liquid/liquid purification of plasma spiked with two different concentrations of RvD1 (1 ng/ml and 0.5 ng/ml), 17S-HDHA (10 ng/ml and 5 ng/ml) and DHA (200 ng/ml and 100 ng/ml).

Product
Concentration ( Table 3 List of primers: primers used for qRT-PCR.