Spontaneous and LPS-induced cytokine production in supernatants of cell cultures isolated from a skin biopsy in a mouse model of acute psoriasis-like dermatitis

Sorokina E.V., Akhmatova E.A., Bisheva I.V., Stolpnikova V.N., Kalinichenko E.O., Skhodova S.A.

I.I. Mechnikov Research Institute of Vaccines and Sera, Moscow, Russia
Postgraduate Education Academy of Federal medical and biological Agency, Moscow, Russia
P.A. Herzen Moscow Cancer Research Institute, Russia

The purpose of the work performed was to obtain data on spontaneous and LPS-stimulated cytokine production in cell culture supernatants isolated from isolated from skin bioptates of C57BL/6 mice in an imiquimod-induced psoriasis model.
Methods. Skin cells were isolated by spontaneous migration and enzymatic dissociation in a collagenase solution. Cells isolated from the skin were cultured for 24 hours in RPMI-1640 (spontaneous cytokine production) or RPMI-1640 medium with the addition of a solution (10 micrograms/ml) of E. coli LPS (induced). Cytokine levels were determined by flow laser cytofluorometry using the Mouse Th1/Th2/Th17 multiplex test system (Antigenix America, USA) on an FC-500 cytometer.
Results. In the focus of imiquimod-induced inflammation, an increase in the spontaneous production of proinflammatory cytokines IL-1, IL-5, IL-17 was observed by 2.4-3.8 times. At the same time, a 1.9-fold inhibition of IFNy synthesis was detected when cultivating skin cells in the presence of bacterial LPS.