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Updated: May 8, 2026
Isolation, Transfection, and Culture of Primary Human Monocytes
Published on: December 16, 2019
Attenuated expression of interferon-β and interferon-λ1 by human alternatively activated macrophages
Ashraf El Fiky1, Roger Perreault, Gwendolyn J McGinnis
1Laboratory of Immunobiochemistry, Division of Bacterial, Parasitic and Allergenic Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, US Food and Drug Administration, 29 Lincoln Drive, Bethesda, MD 20892-4555, USA.
Abstract:
Macrophages can be polarized into classically (CAM) or alternatively (AAM) activated macrophages with IFN-γ or IL-4, respectively. CAM are associated with type 1 immune responses and are implicated in autoimmunity; AAM are associated with type 2 responses and are implicated in allergic diseases. An impediment in investigating macrophage biology using primary human monocyte derived macrophages is the wide inter-donor heterogeneity and the limited quantity of cells that survive in vitro polarization. To overcome this impediment, we established a protocol to generate CAM and AAM cultures derived from the THP-1 human promonocytic cell line. In this report, we demonstrate that THP-CAM and -AAM express gene and protein markers that define their primary human monocyte derived counterparts, such as IL-1β, CXCL10, and CXCL11 for CAM, and MRC1, IL-4 and CCL22 for AAM. In addition, we demonstrate that STAT6 is selectively activated in THP-AAM which, upon LPS stimulation, have an attenuated or delayed expression of IFN-β, IFN-λ1, and IFN α/β pathway genes compared to their CAM counterparts. Taken together, these findings may help further investigate human diseases associated with the alternatively activated macrophage phenotype using this reproducible in vitro macrophage model.
Insights
Researchers developed a reproducible method to create classically (CAM) and alternatively (AAM) activated macrophages from the THP-1 cell line. This model overcomes limitations of primary cells for studying immune responses and diseases like autoimmunity and allergies.
Area of Science:
- Immunology
- Cell Biology
Background:
- Macrophages polarize into classically activated (CAM) and alternatively activated (AAM) phenotypes, crucial for type 1/type 2 immune responses.
- Primary human monocyte-derived macrophages exhibit inter-donor variability and limited in vitro survival, hindering research.
- CAM are linked to autoimmunity, while AAM are implicated in allergic diseases.
Purpose of the Study:
- To establish a reproducible protocol for generating CAM and AAM from the THP-1 human promonocytic cell line.
- To validate THP-1 derived macrophages as a reliable model for studying macrophage polarization.
- To overcome limitations associated with primary human monocyte-derived macrophages.
Main Methods:
- THP-1 cells were differentiated and polarized into CAM (using IFN-γ) and AAM (using IL-4).
- Gene and protein expression of key markers (e.g., IL-1β, CXCL10, MRC1, CCL22) were analyzed.
- STAT6 activation and response to LPS stimulation, including IFN pathway gene expression, were assessed.
Main Results:
- THP-1 derived CAM and AAM expressed characteristic markers of their primary counterparts.
- STAT6 was selectively activated in THP-AAM.
- THP-AAM showed attenuated/delayed IFN-β, IFN-λ1, and IFN α/β pathway gene expression upon LPS stimulation compared to THP-CAM.
Conclusions:
- The THP-1 cell line provides a reproducible in vitro model for CAM and AAM.
- This model facilitates the investigation of human diseases associated with alternatively activated macrophages.
- The findings support the use of THP-1 derived macrophages for studying macrophage biology and immune responses.
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