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TDK conference 2023Hanyecz Viktória - year 4 University of Veterinary Medicine Budapest, Department of Physiology and Biochemistry Supervisor: Csilla Papp-Sebők One of the most defining problems of our days is antimicrobial resistance, since multidrug resistant pathogens are a huge threat both in the field of human and veterinary medicine. There are many initiations regarding the reduction of the usage of antibiotics in the veterinary field, although it's not enough for the curation of the issue, studying and developing antibiotic alternatives are also essential for long term solution. One of the possible directions of these researches is studying antimicrobial peptides (AMPs). AMPs are oligopeptides which either can directly combat microbes or may affect the immune system to successfully eliminate them. In this study the effects of a natural AMP, chicken cathelicidin-2 (Cath-2) were examined on a chicken intestinal epithelial explant model. Throughout the study the freshly isolated cells of the intestine of the broiler chicken were exposed to lipoteichoic acid (LTA) extracted from Staphylococcus aureus to induce inflammation and received treatment of Cath-2 in 5, 10 and 25 nmol/ml concentrations. CCK-8 test were used to assess the metabolic activity, and membrane integrity was studied by measuring the activity of the extracellular lactate dehydrogenase enzyme (LDH). The concertation of proinflammatory interferon gamma (INF-γ) and anti-inflammatory interleukin-10 (IL-10) were measured using Luminex method, and for the purpose of investigating the extent of the inflammation the ratio of these cytokines was calculated. Based on our results Cath-2 did not change either the metabolic activity or the integrity of the membrane of the cells. Furthermore, it was shown that in the groups treated with LTA the concentration of INF-γ was significantly elevated compared to the control group without LTA treatment, although Cath-2 did not cause a decrease compared to the groups with LTA treatment. The concentration of IL-10 significantly decreased in the groups with LTA, LTA+5 nmol/ml Cath-2 and LTA+10 nmol/ml Cath-2 treatment. In addition, the treatment of LTA+25 nmol/ml Cath-2 significantly increased the concentration of IL-10 compared to the group treated solely with LTA. Based on the ratio of INF-γ and IL-10 concentration it could be concluded that all the groups treated with LTA showed a significantly higher quotient compared to the control group without LTA, but it was also found that the treatment together with Cath-2 in the concentration of 25 nmol/ml significantly reduced the ratio compared to the group with only LTA treatment. Based on the results it can be stated that the studied AMP has significant immunomodulatory effects, since it was able to balance out the IL-10 reductant effect of LTA, furthermore it is seen from the ratio of INF-γ and IL-10 that it can successfully moderate the inflammation induced by LTA. Considering these results, chicken cathelicidin-2 can be considered a potential candidate for moderating the various damaging effects of bacterial infections. List of lectures |