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Characterization of <em>In vitro</em> Generated Human Polarized Macrophages | OMICS International | Abstract
ISSN: 2155-9899

Journal of Clinical & Cellular Immunology
Open Access

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Research Article

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.