Abstract
Discovery of novel immune biomarkers for monitoring of disease prognosis and response to therapy in immune-mediated inflammatory diseases is an important unmet clinical need. Here, we establish a novel framework for immunological biomarker discovery, comparing a conventional (liquid) flow cytometry platform (CFP) and a unique lyoplate-based flow cytometry platform (LFP) in combination with advanced computational data analysis. We demonstrate that LFP had higher sensitivity compared to CFP, with increased detection of cytokines (IFN-γ and IL-10) and activation markers (Foxp3 and CD25). Fluorescent intensity of cells stained with lyophilized antibodies was increased compared to cells stained with liquid antibodies. LFP, using a plate loader, allowed medium-throughput processing of samples with comparable intra- and inter-assay variability between platforms. Automated computational analysis identified novel immunophenotypes that were not detected with manual analysis. Our results establish a new flow cytometry platform for standardized and rapid immunological biomarker discovery with wide application to immune-mediated diseases.
Highlights
Chronic inflammation and dysregulated activation of the immune system are central to the pathogenesis of immunemediated inflammatory diseases (IMID), such as psoriasis, rheumatoid arthritis, and Crohn’s disease [1,2,3,4]
We report that lyoplate-based flow cytometry platform (LFP) showed higher sensitivity to detect key cytokines (IFN-c and IL-10) and activation markers (Foxp3 and CD25)
Flow Cytometry antibody cocktail and lyoplate design Our goal was to assess the performance of a lyoplate-based flow cytometry platform (LFP) versus a conventional flow cytometry platform (CFP) using a panel of 12 parameters, including surface markers (CD3, CD4, CD8, CD45RO, and CD25) and intracellular markers (IFN-c, IL-10, IL-17A, and Foxp3) focussing on the investigation of T cell subsets [T helper (Th) 1, Th17, T regulatory cells (T regs) and CD8+ T cells]
Summary
Chronic inflammation and dysregulated activation of the immune system are central to the pathogenesis of immunemediated inflammatory diseases (IMID), such as psoriasis, rheumatoid arthritis, and Crohn’s disease [1,2,3,4]. Given the impact on quality of life, productivity, and the high medical costs related to IMID, the need to understand disease immunopathogenesis is accompanied by an urgent demand to identify specific biomarkers for the purpose of disease screening, diagnosis, staging, and monitoring, as well as to evaluate therapy response. High-dimensional flow cytometry has emerged as a suitable tool for the identification of immunological biomarkers, relevant for IMID and for cancer [7], cardiovascular disease [8], allograft rejection and tolerance [9,10], and infectious diseases [11]. Multicolour flow cytometry has become one of the preferred tools to study the immune system, allowing the simultaneous characterization of many cell types and their functions in complex tissue compartments such as blood, opening the way to a faster and more sophisticated biomarker discovery in human immunology. There is a clear need for a more sophisticated standardized diagnostic platform
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
