Immunomodulatory effects of M. canis on murine epidermal cells.

BURSTEIN V. L., MASIH D. T., CHIAPELLO L

Los Cocos

Congreso; LXI Reunión Científica Anual de la Sociedad Argentina de Inmunología. SAI 2013.; 2013

Sociedad Argentina de Inmunología

Langerhans cells (LC) are the only dendritic cells (DC) in the epidermis (MHC-II+) and represent 1-5% of all nucleated cells in this skin layer. Microsporum canis is a dermatophytic fungus that invades epidermis causing cutaneous mycosis. The effects of dermatophytes on the activation of skin DC as well as the impact on specific antifungal immunity remain unknown. The aim of this work was to study in vitro effects of M. canis on the activation of epidermal cells and to evaluate different isolation and purification methods of murine LC. We studied different sources of epidermal cells from BALB/c mice ears: total epidermal cell (TEpC) suspensions and migratory epidermal cells (MEpC). Regarding TEpC suspensions, we found 2,2 ± 0,3% MHC-II+ cells by flow cytometry. After 24 h- culture, M. canis hyphae (suspension of DO:0,508 ± 0,002, corresponding to 1110 ± 101 colony forming units) induced a significantly increase in IL-6 (p<0,05) and reduced levels of IL-10 production (p<0,004) by TEpC (5x105/well/300l), however there was no difference in MHC-II expression compared to controls. We next examined MEpC by culturing epidermal sheets treated or not with GM-CSF or M. canis, for 3 days. MEpC alone showed 6% MHC-II+ cells, including a 2,4% MHC-IIhi population. Both GM-CSF and M. canis increased the percentage (p<0,00006; p<0,0006, respectively) and the MFI (p<0,01; p<0,003, respectively) of MHC-IIhi cells. Moreover, M. canis induced an increase of IL-6 production. Finally, we assessed LC purification through immunomagnetic beads and isolated 67,7 ± 1,9% MHC-II+ cells with a viability of 70% and a 2% recovery rate, approximately. Furthermore, we evaluate LC isolation by cell sorting with an anti MHC-II antibody and obtained 75% MHC-II+ cells, with 50% viability and a recovery rate of 5%, approximately. Thus, this work set the basis to study murine LC activation due to M. canis in the epidermis microenvironment and directly on isolated LCs.