Characterization of In vitro Generated Human Polarized Macrophages
|Salma Iqbal1 and Ashok Kumar1,2,3*|
|1Department of Pathology and Laboratory Medicine, University of Ottawa, Ottawa, Ontario, Canada|
|2Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, Ontario, Canada|
|3Research Institute, Children’s Hospital of Eastern Ontario, University of Ottawa, Ottawa, Ontario, Canada|
|Corresponding Author :||Ashok Kumar
Division of Virology
Research Institute, Children’s Hospital of Eastern Ontario
University of Ottawa, 401 Smyth Road
Ottawa, Ontario, K1H 8L1, Canada
Tel: 613-737-7600 ext. 3920
Fax: 613-738- 4825
E-mail: [email protected]
|Received October 02, 2015; Accepted December 21, 2015; Published December 28, 2015|
|Citation: Iqbal S, Kumar A (2015) Characterization of In vitro Generated Human Polarized Macrophages. J Clin Cell Immunol 6:380. doi:10.4172/2155-9899.1000380|
|Copyright: © 2015 Iqbal S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.|
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Objective: Contact with invading pathogens and/or tissue injury leads to the polarization of macrophages into either a M1 or a M2 state which is further divided into M2a, M2b and M2c subsets. The human macrophage subsets have been poorly characterized. The present study was undertaken to characterize macrophage polarization using a non-exhaustive panel of surface markers with respect to M1, M2a, M2b and M2c macrophages and production of pro- and anti-inflammatory cytokines in response to various toll-like receptors (TLR), ligands.
Methods: We generated various macrophage subsets by treating monocyte-derived macrophages (MDMs) with IFNγ (M1), IL-4 (M2a), LPS and IL-1β (M2b) or IL-10 (M2c) followed by stimulation with toll-like receptor (TLR)- 2, TLR-3 and TLR-4 agonists and analysis of surface marker and cytokines expression was carried out by flow cytometry and ELISA, respectively.
Results: M2a subset was characterized by CD14low, CD163low and TLR4low phenotype and produced high levels of IL-10. M2b subset was characterized by CD14high, CD80high and CD200Rlow phenotype and produced IL-6 prior to stimulation. M2c subset displayed a CD86low, CD163high phenotype and produced high levels of IL-10. M1 subset was characterized by CD80high, CD86high, CD163low and TLR4high phenotype and produced high levels of proinflammatory IFN-g, IL-12, TNFα and IL-23 following stimulation.
Conclusion: This study characterizes all four polarization states in human macrophages. Each polarization state demonstrated a unique cell surface marker profile and cytokine profile. These phenotypic markers can be used to characterize macrophage populations in tissue inflammatory disease conditions in vivo to further understand disease pathogenesis.