Role of IL-18 in MI-Induced Myelopoiesis

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2022-05

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The Ohio State University

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Interleukin-18 (IL-18), a prominent member of the IL-1 pro-inflammatory cytokine family, has been found to be upregulated via inflammasome signaling post-myocardial infarction (MI), much like its sibling cytokine IL-1β. Contemporary understanding of IL-1β far surpasses that of IL-18 in the context of neutrophil signaling for myelopoiesis post-MI. IL-18 and IL-1β share a myriad of characteristics ranging from analogous protein structure to undergoing the same inflammasome processing. A pivotal study presented by Sreejit et al 2022 mechanistically detailed reverse neutrophil migration and delivery of IL-1β to the bone marrow (BM) to upregulate myelopoiesis post-MI. The objective of this thesis is to explore the potential of IL-18 to be secreted and induce myelopoiesis, similar to IL-1β. To address clinical relevance of this study, the CANTOS Trial displayed promising therapeutic potential of IL-1β blockade in cardiovascular disease (CVD), thus the information this study uncovers concerning IL-18 may have potential to be therapeutically valuable. This study first analyzed IL-18 compared against IL-1β secretion and mRNA expression in inflammasome activated neutrophils, recapitulating previous findings regarding IL-1β secretion from neutrophils and revealing novel finding about IL-18 secretion. mRNA expression of inflammasome components and IL-18 and IL-1β was measured in activated neutrophils to determine inflammasome activity as well as cytokine expression. The potential for IL-18 to induce myelopoiesis in bone marrow derived stem cells was assessed using fluorescent labeling of newly synthesized DNA. All assays were performed in vitro on murine cell cultures and their products. Classical inflammasome priming and subsequent activation, with lipopolysaccharide (LPS) from E. coli and Nigericin, produced upregulated IL-1β mRNA expression and secretion, but unexpectedly failed to produce the same results with IL-18. Confirmations of assay efficacy were performed to validate these results. Proliferaton was found to be highest in the unstimulated cells compared to IL-18 and LPS stimulated cells demonstrating that IL-18 alone is not sufficient in order to stimulate myelopoiesis. IL-18 may exhibit a unique secretion and expression pattern compared to IL-1β. These results also suggest that IL-18, isolated from other stimulating factors, is incapable of stimulating myelopoiesis in an in vitro setting.

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