Leishmania spp. ENFEKSİYONLARININ MAKROFAJ POLARİZASYONU ÜZERİNDEKİ ETKİLERİNİN ARAŞTIRILMASI
Project no. 29627 · Status: Kapanmış · 2023
Summary
<p>Leishmania spp. infection varies in species and subspecies and affects the activation and phenotype of infected macrophages. In our study, we aimed to determine the effect of Leishmania tropica (L. tropica), the main causative agent of cutaneous leishmaniasis in Turkey, on macrophage polarization.</p><p>In the project, the phenotypes of macrophages were evaluated using genetic, metabolic and immunological markers in an ex-vivo infection model in which macrophages generated by induction of human monocytes were infected with the local L. tropica strain. The expression levels of macrophage (CD11b), M1 (CD80) and M2 (CD163, CD209 and CD206) surface markers were analyzed by flow cytometry and changes in the expression of related genes (ARG-1, NOS2a, IDO1, IRF4, CCL1 and CCL22) by RT-qPCR. In addition, the amount of nitric oxide (NO) released outside the cell as a result of infection and Arginase activity were also determined.</p><p>Flow cytometry analysis showed a significant decrease in the percentage of the M1 (classical activation) marker CD80 and an increase in the percentage of the M2 (alternative activation) marker CD163 in infection. Other M2 markers CD206 and CD209 showed decreased percentages. A significant increase in the expression of ARG1 gene associated with M2 phenotype and a significant decrease in the expression of NOS2a and IDO1 genes associated with M1 phenotype were observed in infection. The expression of the M2-related IRF4 gene was also significantly decreased. Changes in CCL1 and CCL22 gene expressions were not statistically significant. The lack of statistically significant changes in NO level and Arginase activity (p value 0, 10) was associated with the fact that they were monitored once in a certain hour, and it is foreseen that they should be evaluated at certain intervals in future studies.</p><p>By analyzing surface markers and gene expression levels, it was observed that infection activated macrophages specific to the M2 phenotype, and the decrease in some M2 markers suggested the role of M2 macrophage subtypes in Leishmania-dependent activation. Therefore, the relationship between L. tropica infection and macrophage subtypes needs further studies.</p>