Morpho-phenotypic Characterisation of Immune Cells Using an Ex vivo Chronic Inflammation Model

Abstract

Autoimmune diseases such as rheumatoid arthritis (RA) often involve prolonged immune dysregulation well before clinical symptoms appear, making timely detection crucial. It can be hypothesised that the morphology of immune cells can function as an early detection marker in an inflammatory system and hence aid in early detection of immune dysregulation. Leveraging this, we developed an ex vivo chronic inflammation model by culturing peripheral blood mononuclear cells (PBMCs) from age-matched RA patients and healthy donors under inflammatory cytokine conditions that replicate the immune environment of early disease. Using the means of high-content imaging, unique morpho-phenotypic changes within immune cells, distinct clustering patterns, and significant shifts in cell size and shape in RA-derived PBMCs compared to healthy controls were observed. We also identified a novel extracellular structure termed “WEBs” that varied markedly between patients and controls. Quantitative analyses of these phenotypic differences revealed measurable variations in immune cell behaviour that could serve as early indicators of autoimmune dysregulation. By integrating AI-driven analysis, our approach systematically detects subtle cellular changes, offering a robust framework for refining diagnostic tools and improving early intervention in RA and other autoimmune diseases.