Katarzyna Swist-Szulik
Newcastle upon Tyne University, UK
Title: The role of mitochondrial dysfunction in infl ammatory crosstalk between immune cells and non-myeloid cells such as fi broblasts and myoblasts
Biography
Biography: Katarzyna Swist-Szulik
Abstract
Introduction: Mitochondria are double-membrane-bound organelles and primary source of cellular energy adenosine triphosphate (ATP). Mitochondria are also involved in innate responses and are necessary for NLRP3 inflammasome activation and maturation of IL-1β in immune cells. Research project investigates the role of mitochondrial dysfunction in cell to cell cytokine crosstalk ((1-6) . Aims: To characterise the nature of cytokine crosstalk between fibroblasts and myoblasts with induced mitochondrial dysfunction and inflammatory cells such as monocytes and THP1 cells. Methods: Stimulation with LPS (lipopolysaccharide) and benzylated ATP were used to cleavage NLRP3 inflammasome and release IL-1β in monocytes and THP1 cells. Supernatants containing IL-1β from treated for inflammasomes THP1 cells as well as recombinant IL-1β were applied on fibroblasts and myoblasts with induced by Rotenone ( Complex I inhibitor) mitochondrial dysfunction. Blocking experiment using anti-IL-1β, anti-IL-1α antibodies as well as IL-1 receptor antagonist characterized the interactions between IL-1β and Il-6 cytokines. ELISA tests were used to measure the content of IL-1β and IL-6 in supernatants. Seahorse was used to assess the changes in bioenergetics profile of the treated cells. Results: Fibroblasts and myoblasts release IL-6 in the presence of supernatants containing IL-1β from THP1 cells and monocytes in the dose dependent manner. Cells with induced mitochondrial dysfunction produce high levels of IL-6 level in close correlation with the degree of mitochondrial dysfunction and dose of recombinant IL-1β. IL-6 released by fibroblasts is in 90% inhibited by adding IL-1 receptor antagonist (IL-Ra) and anti-IL-1β antibodies what would suggest single cytokine crosstalk between IL-1β and IL-6. Conclusions: The data indicate the presence of crosstalk between monocytes and fibroblasts or myoblasts on the cytokine level characterized by IL-1β and IL-6 relationship. Ability to produce high levels of IL-6 by fibroblasts or myoblasts is amplified by degree of mitochondrial dysfunction and presence of IL-1β in dose dependent manner.