Journal Article

Effect of stimulation time on the expression of human macrophage polarization markers

Unuvar Purcu, Duygu, Helvaci, Derya Goksu, Dogan, Yavuz, Sag, Duygu, Korkmaz, Asli, Gunalp, Sinem, Wingender, Gerhard, Erdal, Yonca, Suner, Asli

2022

Overview

Macrophages are highly plastic cells that can polarize into functionally distinct subsets in vivo and in vitro in response to environmental signals. The development of protocols to model macrophage polarization in vitro greatly contributes to our understanding of macrophage biology. Macrophages are divided into two main groups: Pro-inflammatory M1 macrophages (classically activated) and anti-inflammatory M2 macrophages (alternatively activated), based on several key surface markers and the production of inflammatory mediators. However, the expression of these common macrophage polarization markers is greatly affected by the stimulation time used. Unfortunately, there is no consensus yet regarding the optimal stimulation times for particular macrophage polarization markers in in vitro experiments. This situation is problematic, (i) as analysing a particular marker at a suboptimal time point can lead to false-negative results, and (ii) as it clearly impedes the comparison of different studies. Using human monocyte-derived macrophages (MDMs) in vitro , we analysed how the expression of the main polarization markers for M1 (CD64, CD86, CXCL9, CXCL10, HLA-DR, IDO1, IL1β, IL12, TNF), M2a (CD200R, CD206, CCL17, CCL22, IL-10, TGM2), and M2c (CD163, IL-10, TGFβ) macrophages changes over time at mRNA and protein levels. Our data establish the most appropriate stimulation time for the analysis of the expression of human macrophage polarization markers in vitro . Providing such a reference guide will likely facilitate the investigation of macrophage polarization and its reproducibility.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Biology / Biology and Life Sciences / Biomarkers - metabolism / Biotechnology / CCL17 protein / CCL22 protein / CD163 antigen / CD86 antigen / Chemokines / Computer and Information Sciences / CXCL10 protein / Cytokines / Engineering and Technology / Flow cytometry / Gene expression / Genetic aspects / Genomes / Health aspects / Histocompatibility antigen HLA / Humans / Inflammation / Interleukin 1 / Interleukin 10 / Interleukin 12 / Interleukin-10 - metabolism / Macrophage Activation / Macrophages / Macrophages - metabolism / Markers / Medicine / Medicine and Health Sciences / Metabolism / Monocytes / mRNA / Polarization / Reproducibility of Results / Research and Analysis Methods / Stimulation / Surface markers / Tumor necrosis factor