Innate immunity cells in a model of acute psoriasis-like inflammation in mice

Abstract

Background. Experimental animal models of psoriasis helped to clarify the functions of inflammatory mediators, to reveal the contribution of innate or adaptive immune mechanisms, keratinocytes to the development and maintenance of inflammation in psoriasis.Aim. To study the subpopulation composition of immune cells of blood, skin, lymphoid organs and compare two methods of isolation of cells from the skin.Materials and methods. The study included 46 mice of the C57BL / 6 line, which were divided into 2 groups: experimental (n = 24) to reproduce a model of acute psoriasis-like dermatitis using imiquimod cream 5 % (62.5 mg / cm2 / day / mouse, 7 days) and control (n = 22). The severity of skin inflammation was assessed on a point scale. On the 7th day, the skin, spleen, lymph nodes, and thymus were examined. To isolate cells from the skin, the method of spontaneous migration and enzymatic dissociation using collagenase was used. The assessment of the subpopulation structure of mononuclear cells (MNCs) was carried out by flow cytometry using monoclonal antibodies against the corresponding antigens (CD3, CD4, CD5, CD8, MHC class II, TCRyδ, CD38, CD80, CD83, CD86, TLR2). Statistical processing was carried out using the winMDI 2.8 software package.Results. It has been shown that both methods of isolation of skin cells are applicable for immunophenotyping of γδ T-lymphocytes, CD86+, CD83+, CD83+CD86+ dendritic cells. A decrease in TLR2 expression on blood cells and an increase in lymph node and skin cells were revealed. There was a marked increase in the number of CD38+ in the lymph nodes, thymus, and an increase in γδ T-lymphocytes in the lymph nodes and blood. The infiltration of γδ T-lymphocytes, CD8+ is shown in the skin and CD38+ cells.Conclusion. Acute psoriasis-like inflammation of mice was accompanied by an increase in the number of γδ T cells in the blood, lymph nodes and skin. Infiltration of the skin by CD8+ and CD38+ cells was observed. Both methods of cell isolation – the method of spontaneous migration and the method of enzymatic dissociation proved to be applicable for further immunophenotyping.