Cytokine storm is a severe, life-threatening condition characterized by systemic inflammation, cell hyperreactivity, and the release of large amounts of cytokines, which are important for intercellular communication and regulation of the immune response. The discovery of new biological markers enables earlier diagnosis, monitoring of disease progression and treatment response, as well as identification of new drugs. The early activation marker of lymphocytes CD69, also plays an essential role in cell proliferation, differentiation, migration, and cytotoxic function. Additionally, it could serve as a suitable marker for disease progression and therapy efficacy in certain chronic and inflammatory conditions. The use of such markers in relevant preclinical in vitro models can enhance the understanding of the complex mechanisms of cytokine storm and facilitate the development of new drugs for its treatment. For this purpose, this master’s thesis focused on testing the immunomodulatory effects of compounds based on their impact on cytokine secretion and CD69 expression. We used Jurkat lymphocyte cells and THP-1 monocyte cells. For all compounds, we first used the MTS assay to confirm they did not negatively affect the metabolic activity of the cells. We found that the ionomycin/phorbol 12-myristate 13-acetate combination is the most suitable for stimulating Jurkat cell to express CD69. Additionally, we determined that lipopolysaccharide is the most appropriate stimulant for inducing cytokine secretion in THP-1 cells. Due to the broader spectrum of cytokine secretion, the THP-1 cell line proved to be a more suitable in vitro model for evaluating the effects of compounds on cytokine secretion, however, lipopolysaccharide-induced level of activation of THP-1 cells was inconsistent. On the other hand, the Jurkat cell line was found to be a more appropriate in vitro model for assessing the effects of compounds on CD69 marker expression. We concluded that both in vitro models responded appropriately to the tested compounds, however, it is important to note that the Jurkat cell model does not respond to glucocorticoids due to a defect in glucocorticoid receptor.
Among the most notable compounds were those from the class of MAP kinase inhibitors. In the future, it will be necessary to ensure consistent level of activation of undifferentiated THP-1 cells or to pre-differentiate them into macrophages.
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