Contributions
Abstract: EP760
Type: E-Poster Presentation
Session title: Hematopoiesis, stem cells and microenvironment
Background
Macrophages are immune cells known for the detection and destruction of bacteria and viruses. In addition, macrophages are vital for a more generalised inflammatory response, providing cell-cell contacts and producing a cocktail of cytokines that stimulate a variety of other immunopathology’s; including wound healing. Importantly, macrophages exhibit plasticity in their specific phenotypes, able to transiently convert between pro or anti-inflammatory subsets of cells along a continuum. For example, the pro-inflammatory M1 subset are heavily involved in immune modulation and the anti-inflammatory M2c subset is central to the erythroblastic island in the bone marrow.
Aims
The aim of our work was to further understand and characterise human macrophages and their polarity using a well characterised in vitro culture model. A multipronged approach of mass spectrometry, flow cytometry and a previously described2 imaging pipeline was utilised to interrogate the cell surface expression and behaviour of cultured cells.
Methods
We compared three populations against our control unstimulated macrophage (MØ) population, including the classically activated “inflammatory” M1 stimulated with interferon-g (IFNg) against the non-classically activated M2a macrophage subset stimulated with interleukin-4 (IL-4) and finally M2c macrophages stimulated with dexamethasone. The glucocorticoid dexamethasone was also applied as a treatment of the inflammatory M1 subset to better understand the specific effects of this steroid on macrophage polarisation to mimic anti-inflammatory steroid treatment for diseases such as COVID19.
Results
In doing this we demonstrate an increase in CD163 and CD206, markers synonymous with anti-inflammatory M2 macrophages when compared with untreated M1 macrophages. We detected specific alterations in cell surface markers and behaviour between the subsets of in vitroproduced macrophages associated with the known phenotypes.
Conclusion
This provides a detailed dissection of the macrophage polarity continuum that is relevant to the macrophage field as a whole and to the current situation with COVID-19. This ex vivo method of recapitulating an aspect of the inflammatory response to disease has the prospect to be a test bed for future drugs and therapeutics.
Keyword(s): Dexamethasone, Macrophage, Proteomics
Abstract: EP760
Type: E-Poster Presentation
Session title: Hematopoiesis, stem cells and microenvironment
Background
Macrophages are immune cells known for the detection and destruction of bacteria and viruses. In addition, macrophages are vital for a more generalised inflammatory response, providing cell-cell contacts and producing a cocktail of cytokines that stimulate a variety of other immunopathology’s; including wound healing. Importantly, macrophages exhibit plasticity in their specific phenotypes, able to transiently convert between pro or anti-inflammatory subsets of cells along a continuum. For example, the pro-inflammatory M1 subset are heavily involved in immune modulation and the anti-inflammatory M2c subset is central to the erythroblastic island in the bone marrow.
Aims
The aim of our work was to further understand and characterise human macrophages and their polarity using a well characterised in vitro culture model. A multipronged approach of mass spectrometry, flow cytometry and a previously described2 imaging pipeline was utilised to interrogate the cell surface expression and behaviour of cultured cells.
Methods
We compared three populations against our control unstimulated macrophage (MØ) population, including the classically activated “inflammatory” M1 stimulated with interferon-g (IFNg) against the non-classically activated M2a macrophage subset stimulated with interleukin-4 (IL-4) and finally M2c macrophages stimulated with dexamethasone. The glucocorticoid dexamethasone was also applied as a treatment of the inflammatory M1 subset to better understand the specific effects of this steroid on macrophage polarisation to mimic anti-inflammatory steroid treatment for diseases such as COVID19.
Results
In doing this we demonstrate an increase in CD163 and CD206, markers synonymous with anti-inflammatory M2 macrophages when compared with untreated M1 macrophages. We detected specific alterations in cell surface markers and behaviour between the subsets of in vitroproduced macrophages associated with the known phenotypes.
Conclusion
This provides a detailed dissection of the macrophage polarity continuum that is relevant to the macrophage field as a whole and to the current situation with COVID-19. This ex vivo method of recapitulating an aspect of the inflammatory response to disease has the prospect to be a test bed for future drugs and therapeutics.
Keyword(s): Dexamethasone, Macrophage, Proteomics