Expression ofprotease-activated receptors PAR-2 has been investigated on neutrophils isolated from atopic dermatitis patient peripheral blood and has been analyzed with index SCORAD. The simultaneous expression of both, PAR-2 receptors and heat shock protein 90 (HSP90) was measured on neutrophils at the first week of acute phase of atopic dermatitis. Methods. Peripheral blood neutrophils were isolated on the Percoll gradient. Cells were stained with antibodies to PAR-2 receptor and HSP90 followed by the flow cytometry. Results. The PAR-2 receptors have been shown to be expressed on the plasma membrane of neutrophils obtained from patients with atopic dermatitis. The percentage of PAR-2+ neutrophils was positive correlated with diseases severity. HSP90 was found to be elevated simultaneously with PAR-2 receptors on neutrophils at the acute phase of atopic relapse. Conclusions. Atopic dermatitis patients have the higher level of PAR-2 receptor expression compared with healthy donors. Percentage of PAR-2+ neutrophils is elevated with the increasing of disease severity. Simultaneous elevation of PAR-2 receptors and HSP90 was found at the first week of acute atopic dermatitis.

HSP90 expression on the neutrophil plasma membrane has been investigated in patients with atopic dermatitis. Methods. Human peripheral blood cells were stained with monoclonal antibodies to the HSP90 and were analyzed using the flow cytometer. Results. Neutrophils obtained from patients with atopic dermatitis have been found to express the more significant level of HSP90 on plasma membrane, and percentage of these cells has been shown to be higherfor atopic patients compared with healthy donors. Both, neutrophil count and HSP90 fluorescence were increased according elevation of index SCORAD for atopic patients. The most significant elevation in percentage of neutrophils expressing HSP90 on plasma membrane has been found during the first and second weeks of acute phase of the disease. We did not found the significant differences for HSP90 expression on neutrophils dependent on IgE level in the blood serum. Conclusion. HSP90 expression is elevated on the neutrophil plasma membrane from atopic dermatitis patients compared with healthy donors. Percentage of these cells were positive correlated with SCORAD index.

В клетках крови больных атопическим дерматитом (АД) исследовали экспрессию белка mTOR методом проточной цитометрии. Установлено, что общий пул белка mTOR в лимфоцитах повышен при АД по сравнению с донорами. Экспрессия белка mTOR в CD4+ и CD8+ Т-лимфоцитах крови в период обострения АД выше, чем у доноров и больных в периоде ремиссии. Оба комплекса киназы mTOR (mTORC1 и mTORC2) участвуют в изменениях субпопуляций лимфоцитов при АД. Активация белков Raptor и Rictor (mTORC1 vs mTORC2) достоверно выше в лимфоцитах больных АД легкой и средней степени тяжести (по индексу SCORAD), чем у доноров. The kinase mTOR expression was studied in blood cells of patients with atopic dermatitis (AD) using flow cytometry. The total level of lymphocyte mTOR protein was increased in AD patients compared with healthy donors. The mTOR protein expression in CD4+ and CD8+ T cells was higher in patients with AD in a relapse than in donors and patients in a remission. Both mTORC1 and mTORC2 complexes formed by mTOR kinase were found to participate in lymphocyte disruption during AD. Activation of Raptor and Rictor proteins (from mTORC1 and mTORC2, respectively) was significantly increased in lymphocytes of patients with mild and moderate AD (according to the SCORAD index).

Цель работы - изучить влияние препаратов цинка на уровень экспрессии белков, ассоциированных со стрессом, в лимфоцитах крови больных атопическим дерматитом (АД) в зависимости от уровня цинка в сыворотке крови. Методы. Кровь брали натощак. Определяли концентрацию цинка в сыворотке крови методом абсобрционной спектроскопии. Лимфоциты выделяли на гралиенте плотности Фиколла-Верографина и окрашивали моноклональными антителами ко внутриклеточным белкам. Интенсивность флуоресценции измеряли на проточном цитометре FACSCalibur по программе SimulSet. Статистический анализ проводили по программе «Биостатистика». Результаты. Показано влияние исходной концентрации цинка в сыворотке крови на показатели сигнальных белков в лимфоцитах. При дефиците цинка уровень ингибитора клеточного цикла р21 в лимфоцитах повышен, а терапия цинк пиритионом снижает уровень экспрессии р21. При дефиците цинка в сыворотке крови больных АД уровень экспрессии белков киназ р38 и JNK был повышен по сравнению с показателями лиц без дефицита цинка, и снижался после применения цинк пиритиона у больных АД. При дефиците цинка у больных АД была повышена экспрессия молекулярных шаперонов DNAJA3, DNAJB6 и DNAJC15 в лимфоцитах. Применение цинк пиритиона снижает экспрессию этих стрессовых белков. Заключение. Дефицит цинка в сыворотке крови больных АД сопряжен с развитием стресса в лимфоцитах периферической крови больных. Добавление цинк пиритиона снижает уровень экспрессии стресс-зависимых белков в лимфоцитах крови больных АД. The aim of the work was to study the effect of zinc pyrithione and zinc oxide on stress-associated protein expression in blood lymphocytes of patients with atopic dermatitis (AD) depending on the serum concentration of zinc. Methods. Blood was collected in fasting state. Blood concentration of zinc was determined by absorption spectroscopy. Lymphocytes were isolated by the Ficoll/verografin density gradient centrifugation and stained with monoclonal antibodies to intracellular proteins. The fluorescence intensity was measured on a FACSCalibur flow cytometer using the SimulSet software. Statistical analysis was performed using the Biostatistic software. Results. The study showed the effect of zinc serum concentration at baseline on stress-dependent proteins in lymphocytes. In zinc deficiency, the content of cell cycle inhibitor p21 in lymphocytes was increased whereas the zinc pyrithione treatment reduced the p21 expression. Serum zinc deficiency in patients with AD was associated with increased expression of kinase proteins p38 and JNK compared to patients without zinc deficiency; in patients with AD, the kinase protein expression decreased after administration of zinc pyrithione. In AD patients with zinc deficiency, the lymphocyte expression of the DNAJA3, DNAJB6 and DNAJC15 molecular chaperones was increased. The zinc pyrithione treatment restricted the expression of these stress proteins. Conclusions. Serum zinc deficiency in patients with AD is associated with the development of stress in peripheral blood lymphocytes. The zinc pyrithione treatment restricts the expression of stress-dependent proteins in lymphocytes of patients with AD.