Abstract
We investigated direct effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the surface properties and cytokine-producing activity of human monocytes/macrophages (Mc/Mphs). The CD14+cells were isolated from peripheral blood of healthy donors by positive magnetic separation. The isolated Mc/Mphs were cultured with lipopolysaccharide (LPS, 1 μg/ml) or without LPS for 24 hours. Membrane expression of CD14, CD16, CD119, CD124, and CD197 molecules was assessed by flow cytometry. The contents of tumor necrosis factor-α (TNFα), interleukin-1β (IL-1β), IL-6 and IL-10 in culture supernatants were determined by the enzyme immunoassay technique. It was found that GM-CSF at a concentration range of 0.01-10 ng/ml did significantly reduce the number of cells expressing CD197 (CC receptor of chemokine 7), without significantly affecting the percentage of CD14+(coreceptor of LPS), CD16+(low-affinity Fc receptor), CD119+(IFNγ receptor) and CD124+(IL-4 receptor) cells. At the same time, GM-CSF reduced the contents of CD197+macrophages, as well as CD14+, CD16+, and CD119+cells among the activated cell population, without significantly altering the number of CD124+cells. It was also shown that GM-CSF (10 ng/ml), was able to enhance production of TNFα and IL-6, but not IL-1β and IL-10 by activated Mc/Mphs. The results obtained indicate the ability of GM-CSF to exert both anti-inflammatory and pro-inflammatory effects upon macrophage cell populations. In general, such effects could contribute to the development of adaptive immunogenesis in peripheral tissues.
Highlights
CD14+ cells were isolated from peripheral blood of healthy donors by positive magnetic separation
Membrane expression of CD14, CD16, CD119, CD124, and CD197 molecules was assessed by flow cytometry
The contents of tumor necrosis factor-α (TNFα), interleukin-1β (IL-1β), IL-6 and IL-10 in culture supernatants were determined by the enzyme immunoassay technique
Summary
Газатова Н.Д., Меняйло М.Е., Малащенко В.В., Гончаров А.Г., Мелащенко О.Б., Морозова Е.М., Селедцов В.И. Исследовали прямые эффекты гранулоцитарно-макрофагального колониестимулирующего фактора (GM-CSF) человека на поверхностные свойства и цитокин-продуцирующую активность моноцитов/макрофагов (Mц/Мф) человека. Содержание фактора некроза опухоли-α (TNFα), интерлейкина-1β (IL-1β), IL-6 и IL-10 в культуральных супернатантах определяли иммуноферментным методом. Установлено, что GM-CSF в диапазоне концентраций 0,01–10 нг/мл заметно снижал среди неактивированных Мц/Мф количество клеток, экспрессирующих CD197 (C-C рецептор хемокина 7), существенно не влияя на процентное содержание CD14+ (корецептор ЛПС), CD16+ (низкоаффинный Fc-рецептор), CD119+ (рецептор IFNγ) и CD124+ (рецептор IL-4) клеток. В то же время среди активированных Мц/Мф GM-CSF снижал содержание не только CD197+ клеток, но также CD14+, CD16+, и CD119+ клеток, существенно не изменяя количество CD124+ клеток. Что GM-CSF в концентрации 10 нг/мл обладал способностью усиливать продукцию активированными Мц/Мф TNFα и IL-6, но не IL-1β и IL-10. Полученные данные указывают на способность GM-CSF оказывать на макрофагальные клетки как антивоспалительное, так и провоспалительное влияние. Gazatova N.D., Meniailo M.E., Malashchenko V.V., Goncharov A.G., Melashchenko O.B., Morozova E.M., Seledtsov V.I
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