Abstract
T-regulatory cells control the strength and duration of immune response and play an important role in the pathogenesis of psoriasis. The goal of the study was to examine the content of T-regulatory cells in peripheral blood and affected skin of psoriasis patients, the level of FOXP3 mRNA expression in psoriatic skin. Determination of subpopulations of T-regulatory lymphocytes in peripheral blood was performed using flow cytometry. The median level of FOXP3 mRNA expression was determined in biopsies of the skin by PCR in real time. Immunohistochemistry was performed with monoclonal anti- FOXP3 antibodies. Revealed an increase in the relative content of T-regulatory cells in the peripheral blood of patients with psoriasis in the progressive period, compared with healthy people. The level of FOXP3 mRNA expression in psoriatic foci was 3.1 times higher than in healthy individuals. Was noted a 10-fold increase in the number of T-regulatory lymphocytes in the skin of psoriasis patients in the progressive period, compared with skin from healthy donors.
Highlights
The goal of the study was to examine the content of T-regulatory cells in peripheral blood and affected skin of psoriasis patients, the level of FOXP3 mRNA expression in psoriatic skin
The median level of FOXP3 mRNA expression was determined in biopsies of the skin by PCR in real time
Immunohistochemistry was performed with monoclonal antiFOXP3 antibodies
Summary
В группу больных вульгарным псориазом вошел 41 пациент в возрасте от 21 года до 79 лет (средний возраст 47,6±2,04 года). Наиболее специфичным маркером для идентификации T-reg клеток является фактор транскрипции FOXP3, но его локализация внутри клетки значительно осложняет идентификацию T-регуляторных лимфоцитов при проточной цитометрии. Для окрашивания клеток использовали моноклональные антитела, меченные FITC (изотиоцианат флуоресцеина), PE (фикоэритрин) и PC5 (комплекс PE с цианином-5), производства «Beckman Coulter» (США): CD4/FITC, CD25/ PC5, CD127/PE. Исследование экспрессии мРНК FOXP3 Материалом для исследования экспрессии мРНК FOXP3 служили биоптаты кожи, полученные методом панч-биопсии (6 мм) от 37 больных псориазом в прогрессирующем периоде из периферии псориатических бляшек и от 16 здоровых доноров. Для иммуногистохимической детекции Т-регуляторных лимфоцитов использовали первичные мышиные моноклональные антитела анти-FOXP3 (FOXP3 antibody [236A/E7], Abcam, США), систему визуализации Envision (Dako, Дания), в качестве хромогена применяли диаминобензидин (Dako, Дания). Полученные данные представлены в виде среднего количества FOXP3-позитивных клеток для каждого биоптата.
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