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TDK conference 2023Ujházy András Dávid - year 5 University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry Supervisor: Dr. Csilla Papp-Sebők The phenomenon of antimicrobial resistance (AMR), caused by the large-scale and irresponsible use of antibiotics, has become a growing problem worldwide. Antimicrobial peptides (AMPs), which are small cationic peptides produced as part of the innate immune system in almost all living organisms, may be a possible solution to this challenge due to their direct antimicrobial and immunomodulatory effects. IDRs (Innate Defense Regulators) are a group of synthetic, small (10-20 amino acid long) peptides that have almost no direct bactericidal activity, but are effective in treating various infectious and inflammatory diseases due to their immunomodulatory activity. In our study, we investigated the immunomodulatory effect of the peptide IDR-1002 on intestinal epithelial explants of chicken origin. Explants were treated with solutions containing different concentrations of IDR-1002 (10-, 30- and 90 µg/ml) or lipoteichoic acid (LTA) (10 µg/ml) or a combination of both for 12 hours. Metabolic activity and extracellular lactate dehydrogenase (LDH) activity, indicative of membrane damage, were determined by photometry. In addition, the concentration of interleukin (IL)-8 was measured by ELISA assay and levels of RANTES, IL-2 and interferon (IFN)-γ were assessed using Luminex xMAP technology. Based on our results, IDR-1002 did not affect cell viability , as metabolic activity was not significantly decreased at any concentration and no increase in LDH activity was found. As for the proinflammatory biomarkers (IFN- γ, RANTES, IL-8) and IL-2, we observed that LTA increased their concentrations, and this elevating effect was attenuated by IDR-1002 treatment with 10 µg/ml concentration for each of these cytokines. In addition, treatment with 30 µg/ml concentration of the peptide was shown to have a reducing effect on RANTES, while the levels of IL-8 were decreased after treatment with 30- and 90 µg/ml IDR-1002. Furthermore, we observed that the AMP alone had an up-regulating effect on IL-2 production compared to the control group. Our results show that while IDR-1002 had no cell damaging effect, it was able to influence LTA-induced inflammation. Moreover, an increased efficacy of lower concentrations of the peptide was observed compared to higher doses, highlighting the importance of selecting the appropriate treatment concentration. Thus, the peptide we investigated may be a suitable and promising tool to mitigate the adverse health effects of various bacterial and inflammatory diseases, and it is therefore important to conduct further research to map its effects as accurately as possible. List of lectures |