Baxter, EW, Graham, AE, Re, NA et al. (4 more authors) (2020) Standardized protocols for differentiation of THP-1 cells to macrophages with distinct M(IFNγ+LPS), M(IL-4) and M(IL-10) phenotypes. Journal of Immunological Methods, 478. 112721. ISSN 0022-1759
Abstract
In vitro models of differing macrophage functions are useful since human monocyte–derived macrophages are short–lived, finite and vary from donor to donor. Published protocols using the promonocytic cell line THP-1 have tended to result in cells that closely resemble classically-activated macrophages, differentiated in IFNγ and LPS. However, no protocol, to date, has fully recapitulated polarization of THP-1 to the M(IL-4) or M(IL-10) macrophage phenotypes seen when human monocyte-derived macrophages are exposed to each cytokine. Here we present protocols that can be used to prepare M(IL-4) polarized THP-1 that transcribe CCL17, CCL26, CD200R and MRC1 and M(IL-10) cells which transcribe CD163, C1QA and SEPP1. We show that the inhibitory Fcγ Receptor IIb is preferentially expressed on the surface of M(IL-4) cells, altering the balance of activating to inhibitory Fcγ Receptors.
Adoption of standardized experimental conditions for macrophage polarization will make it easier to compare downstream effector functions of different macrophage polarization states, where the impact of PMA exposure is minimized and rest periods and cytokine exposure have been optimized.
Metadata
Item Type: | Article |
---|---|
Authors/Creators: |
|
Copyright, Publisher and Additional Information: | © 2019 Published by Elsevier B.V. This is an author produced version of a paper published in Journal of Immunological Methods. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | THP-1; Differentiation; Macrophage polarization; M(IFNγ+LPS); M(IL-4); FcγRIIb |
Dates: |
|
Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Medicine and Health (Leeds) > School of Medicine (Leeds) > Leeds Institute of Cardiovascular and Metabolic Medicine (LICAMM) > Discovery & Translational Science Dept (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 03 Jan 2020 12:33 |
Last Modified: | 04 Feb 2021 01:39 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.jim.2019.112721 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:155057 |
Downloads
Filename: BaxteretalJIM final (2).pdf
Licence: CC-BY-NC-ND 4.0
Filename: Graphical Abstract.tiff
Licence: CC-BY-NC-ND 4.0
Filename: Figure1 (1).tiff
Licence: CC-BY-NC-ND 4.0
Filename: Figure2.tiff
Licence: CC-BY-NC-ND 4.0
Filename: Figure3.tiff
Licence: CC-BY-NC-ND 4.0
Filename: Figure4.tiff
Licence: CC-BY-NC-ND 4.0
Filename: Revised Figure 5.tiff
Licence: CC-BY-NC-ND 4.0
Filename: Supplementary Figure 1.tiff
Licence: CC-BY-NC-ND 4.0
Filename: Supplementary Figure 2.tiff
Licence: CC-BY-NC-ND 4.0
Filename: Supplementary Figure 3a.tiff
Licence: CC-BY-NC-ND 4.0
Filename: Supplementary Figure 3b (1).tiff
Licence: CC-BY-NC-ND 4.0