My research project was focused on validation of a multicytokine molecular panel by Kairos automated system recently developed by Nurex srl in patients with hypereosinophilic syndromes, a group of rare immunohematologic disorders. Kairos multicytokine molecular panel requires mRNA extraction, purification, followed by multicytokines mRNA quantification. As preliminary step, the multicytokine panel was tested in a model of acute inflammatory response such as sepsis and then customized and transferred to a model of chronic inflammation such as the hypereosinophilic syndrome. Hypereosinophilic syndromes are a heterogeneous group of disorders characterized by eosinophil-driven inflammation and complex cytokine dysregulation. These conditions pose diagnostic and therapeutic challenges due to their distinct pathogenetic mechanisms and often overlapping clinical presentations. A 14-cytokine multiplex RT-qPCR assay panel was designed, encompassing key players in the pathogenesis of hypereosinophilic syndromes and cytokines involved in inflammation and tissue remodeling (IL-4, IL-5, IL-13, IL-33, TSLP, IL-17A, TGF-β, TNF-α, IL-1B, IL-6, IL-10, IL-1ra, IL-2, and IFN-γ). The specificity, sensitivity, and ability of the platform to monitor dynamic cytokine changes were previously validated. Notably, the platform requires only 25-100µl of blood and completes the entire process in approximately 2 hours, overcoming limitations of traditional methods. This study aimed to develop a novel automated multicytokine platform for rapid and robust cytokine profiling in whole blood samples of patients with various hypereosinophilic syndromes, addressing the need for accurate and efficient diagnostic tools. The platform was used to analyze whole blood samples from 63 patients with hypereosinophilic syndromes. Our multicytokine panel was able to identify a distinct cytokine signature between hypereosinophilic syndromes and sepsis, systemic hypereosinophilic syndromes and sepsis, localized and systemic hypereosinophilic syndromes, and subgroups of systemic hypereosinophilic conditions, highlighting its potential for improving diagnostic accuracy and guiding personalized treatment.

Exploring The Role of a Novel Highly Automated Technology for Multipanel Cytokines Determination in Hypereosinophilic Syndromes

Ribeiro Luz Soraia
2025-01-01

Abstract

My research project was focused on validation of a multicytokine molecular panel by Kairos automated system recently developed by Nurex srl in patients with hypereosinophilic syndromes, a group of rare immunohematologic disorders. Kairos multicytokine molecular panel requires mRNA extraction, purification, followed by multicytokines mRNA quantification. As preliminary step, the multicytokine panel was tested in a model of acute inflammatory response such as sepsis and then customized and transferred to a model of chronic inflammation such as the hypereosinophilic syndrome. Hypereosinophilic syndromes are a heterogeneous group of disorders characterized by eosinophil-driven inflammation and complex cytokine dysregulation. These conditions pose diagnostic and therapeutic challenges due to their distinct pathogenetic mechanisms and often overlapping clinical presentations. A 14-cytokine multiplex RT-qPCR assay panel was designed, encompassing key players in the pathogenesis of hypereosinophilic syndromes and cytokines involved in inflammation and tissue remodeling (IL-4, IL-5, IL-13, IL-33, TSLP, IL-17A, TGF-β, TNF-α, IL-1B, IL-6, IL-10, IL-1ra, IL-2, and IFN-γ). The specificity, sensitivity, and ability of the platform to monitor dynamic cytokine changes were previously validated. Notably, the platform requires only 25-100µl of blood and completes the entire process in approximately 2 hours, overcoming limitations of traditional methods. This study aimed to develop a novel automated multicytokine platform for rapid and robust cytokine profiling in whole blood samples of patients with various hypereosinophilic syndromes, addressing the need for accurate and efficient diagnostic tools. The platform was used to analyze whole blood samples from 63 patients with hypereosinophilic syndromes. Our multicytokine panel was able to identify a distinct cytokine signature between hypereosinophilic syndromes and sepsis, systemic hypereosinophilic syndromes and sepsis, localized and systemic hypereosinophilic syndromes, and subgroups of systemic hypereosinophilic conditions, highlighting its potential for improving diagnostic accuracy and guiding personalized treatment.
2025
RT-qPCR
automated mRNA extraction, purification
Automated platform
Biomarkers
Th2-related diseases
Molecular diagnostic
Sepsis biomarkers
Hypereosinophilic syndromes Biomarkers
Hypereosinophilic Syndromes
Cytokines
Cytokine profiling
Inflammation
Immune dysregulation
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Descrizione: This doctoral thesis in biomolecular medicine presents research on the development and validation of a novel highly automated platform for rapid and robust mRNA multicytokine profiling in hypereosinophilic syndromes.
Tipologia: Tesi di dottorato
Licenza: Creative commons
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11562/1152169
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