Comparative Proteomic Analysis of Polarized Human THP-1 and Mouse RAW264.7 Macrophages

Abstract

Macrophages can be polarized into classically activated macrophages (M1) and alternatively activated macrophages (M2) in the immune system, performing pro-inflammatory and anti-inflammatory functions, respectively. Human THP-1 and mouse RAW264.7 cell line models have been widely used in various macrophage-associated studies, while the similarities and differences in protein expression profiles between the two macrophage models are still largely unclear. In this study, the protein expression profiles of M1 and M2 phenotypes from both THP-1 and RAW264.7 macrophages were systematically investigated using mass spectrometry-based proteomics. By quantitatively analyzing more than 5,000 proteins among different types of macrophages (M0, M1 and M2) from both cell lines, we identified a list of proteins that were uniquely up-regulated in each macrophage type and further confirmed 43 proteins that were commonly up-regulated in M1 macrophages of both cell lines. These results revealed considerable divergences of each polarization type between THP-1 and RAW264.7 macrophages. Moreover, the mRNA and protein expression of CMPK2, RSAD2, DDX58, and DHX58 were strongly up-regulated in M1 macrophages for both macrophage models. These data can serve as important resources for further studies of macrophage-associated diseases in experimental pathology using human and mouse cell line models.

Nitric Oxide Induced stx2 Expression Is Inhibited by the Nitric Oxide Reductase, NorV, in a Clade 8 Escherichia coli O157:H7 Outbreak Strain

“…There is also the possibility that the RAW264.7 mouse derived macrophage cells, which were used to examine the survival of the EHEC K15 strain after 10 h [18], may differ from the human derived THP-1 cells used herein. Indeed, a prior study observed key differences in protein expression profiles between mouse-derived RAW264.7 macrophages and THP-1 cells [43].…”

Construction of a Macrophage Infiltration Regulatory Network and Related Prognostic Model of High-Grade Serous Ovarian Cancer

“…We selected the genes to construct the macrophage infiltration network, in which seven genes with high connectivity played a leading role in this network. RSAD2 is an antiviral protein that is significantly upregulated in M1 macrophages [ 29 ]. IFIT3 is a marker of M1 macrophage polarization and is highly upregulated in atherosclerosis and other inflammatory diseases [ 30 ].…”

Mesenchymal-Stromal Cell-like Melanoma-Associated Fibroblasts Increase IL-10 Production by Macrophages in a Cyclooxygenase/Indoleamine 2,3-Dioxygenase-Dependent Manner

“…Macrophage M2-like polarization was achieved by 72 h incubation with 20 ng/mL of interleukin 4 (PeproTech) and 20 ng/mL of interleukin 13 (PeproTech). M1-like CD38 [ 22 , 23 ] and M2-like and CD209 [ 23 , 24 ] marker expression on macrophages was confirmed with flow cytometry (BD FACSCalibur™ system, BD Biosciences, San Diego, CA, USA). Plots were analyzed with FlowJo software ( Figure 1 d).…”