Macrophages - the most abundant antigen-presenting cells in the periphery - represent by themselves a wide group of functionally-different subpopulations involved in several immunological functions, including inflammatory responses and tissue homeostasis maintain & repair. Unlike M1 macrophages which promote a pro-inflammatory response, alternatively activated M2 macrophages, with diametrically opposed functions, trigger inflammation resolution and suppress T cell activation via several mechanisms including immune checkpoint engagement (e.g. PDL1), release of anti-inflammatory cytokines such as IL-10 and TGFb, and metabolic activities which promote essential amino acid depletion (e.g. Arginine). – which characteristics that they share with the so-called tumor-associated macrophages (TAMs).
Our newly validated M2 suppression assay based on i) the co-culture of autologous monocyte-derived M2 macrophages and activated CD4+ T cells (or PBMCs) and on ii) the quantitation of IFNg levels as surrogate of T cell activation, is specifically designed to assess new immunotherapeutics for their modulatory activity on the phenotype and function of M2 macrophages. Candidate compounds can thus be evaluated as single agents or in combinatorial treatments, for their potential to repolarize / switch M2 macrophages and to antagonize M2-mediated T cell suppression.
Molecular features of human monocyte-derived macrophages through surface marker expression and cytokine profiles

While displaying an expected variation from a donor to another one, surface marker expression ...
Assay principle
- HTRF-based quantification of key cytokines released by macrophages, e.g. IL-6, IL-10, IL-12 as to evaluate the potency of candidate compounds to revert M2 phenotype.
- Human autologous M2 suppression assay, based on a co-culture of monocyte-derived M2 macrophages with activated T cells (or PBMCs) and measurement of IFNγ, as one of the most representative surrogates of T cell activation.
- Robust functional assay, microplate format tailored for screening campaigns
- Complementary analysis can be implemented to decipher MOA (e.g. activation markers, signaling pathways, etc)