mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-2019-3-419-426mimmun-1815Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESПРЯМЫЕ ЭФФЕКТЫ ГРАНУЛОЦИТАРНО-МАКРОФАГАЛЬНОГО КОЛОНИЕСТИМУЛИРУЮЩЕГО ФАКТОРА НА ФУНКЦИОНАЛЬНЫЕ СВОЙСТВА МОНОЦИТОВ/МАКРОФАГОВ ЧЕЛОВЕКАDIRECT EFFECTS OF GM-CSF ON THE FUNCTIONS OF HUMAN MONOCYTES/MACROPHAGESГазатоваН. Д.GazatovaN. D.

научный сотрудник центра медицинских биотехнологий

г. Калининград

 

Research Associate, Center of Medical Biotechnologies

Kaliningrad

МеняйлоМ. Е.MeniailoM. E.

младший научный сотрудник центра медицинских биотехнологий 

г. Калининград

Junior Research Associate, Center of Medical Biotechnologies

Kaliningrad

МалащенкоВ. В.MalashchenkoV. V.

младший научный сотрудник центра медицинских биотехнологий 

г. Калининград

Junior Research Associate, Center of Medical Biotechnologies

Kaliningrad

ГончаровА. Г.GoncharovA. G.

к.м.н., директор института живых систем 

г. Калининград

PhD (Medicine), Director, Institute of Live Systems

Kaliningrad

МелащенкоО. Б.MelashchenkoO. B.

научный сотрудник центра медицинских биотехнологий 

г. Калининград

Research Associate, Center of Medical Biotechnologies

Kaliningrad

МорозоваЕ. М.MorozovaE. M.

младший научный сотрудник центра медицинских биотехнологий 

г. Калининград

Junior Research Associate, Center of Medical Biotechnologies

Kaliningrad

СеледцовВ. И.SeledtsovV. I.

д.м.н., профессор, главный научный сотрудник центра медицинских биотехнологий

236001, г. Калининград, ул. Гайдара, 6, каб. 406

Тел.: 8 (915) 263-60-27

PhD, MD (Medicine), Professor, Chief Research Associate, Center of Medical Biotechnologies

236001, Kaliningrad, Gaidar str., 6-406

Phone: 7 (915) 263-60-27

seledtsov@rambler.ruФГАОУ «Балтийский федеральный университет имени Иммануила Канта»РоссияI. Kant Baltic Federal UniversityRussian Federation201911072019213419426Copyright © Газатова Н.Д., Меняйло М.Е., Малащенко В.В., Гончаров А.Г., Мелащенко О.Б., Морозова Е.М., Селедцов В.И., 20192019Газатова Н.Д., Меняйло М.Е., Малащенко В.В., Гончаров А.Г., Мелащенко О.Б., Морозова Е.М., Селедцов В.И.Gazatova N.D., Meniailo M.E., Malashchenko V.V., Goncharov A.G., Melashchenko O.B., Morozova E.M., Seledtsov V.I.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.mimmun.ru/mimmun/article/view/1815

Исследовали прямые эффекты гранулоцитарно-макрофагального колониестимулирующего фактора (GM-CSF) человека на поверхностные свойства и цитокин-продуцирующую активность моноцитов/макрофагов (Mц/Мф) человека. CD14+ клетки были выделены из крови здоровых доноров методом позитивной магнитной сепарации. Выделенные Mц/Мф культивировали с липополисахаридом (ЛПС, 1 мкг/мл) или без ЛПС в течение 24 ч. Мембранную экспрессию молекул СD14 CD16, CD119, CD124, CD197 оценивали методом проточной цитофлуориметрии. 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. Также показано, 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. В целом такое влияние может способствовать развитию адаптивного иммуногенеза на периферии.

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.

гранулоцитарно-макрофагальный колониестимулирующий фактормоноцит/макрофагСD-экспрессияцитокиныgranulocyte-macrophage colony-stimulating factormonocyte/macrophagesСD-expressioncytokinesReferencesКудрявцев И.В. Т-клетки памяти: основные популяции и стадии дифференцировки // Российский иммунологический журнал, 2014. Т. 8 (17), № 4. С. 947-964.Kudryavtsev I.V. Memory T cells: major populations and stages of differentiation. Rossiyskiy immunologicheskiy zhurnal = Russian Journal of Immunology, 2014, Vol. 8 (17), no. 4, pp. 947-964. (In Russ.)Becher B., Tugues S., Greter M. GM-CSF: from growth factor to central mediator of tissue inflammation. Immunity, 2016, Vol. 45, no. 5, pp. 963-973.Becher B., Tugues S., Greter M. GM-CSF: from growth factor to central mediator of tissue inflammation. Immunity, 2016, Vol. 45, no. 5, pp. 963-973.Broughton S.E., Nero T.L., Dhagat U., Kan W.L., Hercus T.R., Tvorogov D., Lopez A.F., Parker M.W. The βc receptor family – Structural insights and their functional implications. Cytokine, 2015, Vol. 74, no. 2, pp. 247-258.Broughton S.E., Nero T.L., Dhagat U., Kan W.L., Hercus T.R., Tvorogov D., Lopez A.F., Parker M.W. The βc receptor family – Structural insights and their functional implications. Cytokine, 2015, Vol. 74, no. 2, pp. 247-258.Codarri L., Gyulveszi G., Tosevski V., Hesske L., Fontana A., Magnenat L., Suter T., Becher B. ROR gamma t drives production of the cytokine GM-CSF in helper T cells, which is essential for the effector phase of autoimmune neuroinflammation. Nat. Immunol., 2011, Vol. 12, no. 6 , pp. 560-567.Codarri L., Gyulveszi G., Tosevski V., Hesske L., Fontana A., Magnenat L., Suter T., Becher B. ROR gamma t drives production of the cytokine GM-CSF in helper T cells, which is essential for the effector phase of autoimmune neuroinflammation. Nat. Immunol., 2011, Vol. 12, no. 6 , pp. 560-567.Croxford A.L., Spath S., Becher B. GM-CSF in neuroinflammation: licensing myeloid cells for tissue damage. Trends Immunol., 2015, Vol. 36, no. 10, pp. 651-662.Croxford A.L., Spath S., Becher B. GM-CSF in neuroinflammation: licensing myeloid cells for tissue damage. Trends Immunol., 2015, Vol. 36, no. 10, pp. 651-662.Darrieutort-Laffite C., Boutet M.A., Chatelais M., Brion R., Blanchard F., Heymann D., le Goff B. IL-1β and TNFα promote monocyte viability through the induction of GM-CSF expression by rheumatoid arthritis synovial fibroblasts. Mediators Inflamm., 2014, 241840. doi: 10.1155/2014/241840.Darrieutort-Laffite C., Boutet M.A., Chatelais M., Brion R., Blanchard F., Heymann D., le Goff B. IL-1β and TNFα promote monocyte viability through the induction of GM-CSF expression by rheumatoid arthritis synovial fibroblasts. Mediators Inflamm., 2014, 241840. doi: 10.1155/2014/241840.El-Behi M., Ciric B., Dai H., Yan Y., Cullimore M., Safavi F., Zhang G.X., Dittel B.N., Rostami A. The encephalitogenicity of T(H) 17 cells is dependent on IL-l- and IL-23-induced production of the cytokine GM-CSF. Nat. Immunol., 2011, Vol. 12, no. 6, pp. 568-575.El-Behi M., Ciric B., Dai H., Yan Y., Cullimore M., Safavi F., Zhang G.X., Dittel B.N., Rostami A. The encephalitogenicity of T(H) 17 cells is dependent on IL-l- and IL-23-induced production of the cytokine GM-CSF. Nat. Immunol., 2011, Vol. 12, no. 6, pp. 568-575.Eshleman E.M., Delgado C., Kearney S.J., Friedman R.S., Lenz L.L. Down regulation of macrophage IFNGR1 exacerbates systemic L. monocytogenes infection. PLoS Pathog., 2017, Vol. 13, e1006388. doi: 10.1371/journal.ppat.1006388.Eshleman E.M., Delgado C., Kearney S.J., Friedman R.S., Lenz L.L. Down regulation of macrophage IFNGR1 exacerbates systemic L. monocytogenes infection. PLoS Pathog., 2017, Vol. 13, e1006388. doi: 10.1371/journal.ppat.1006388.Geginat J., Sallusto F., Lanzavecchia A. Cytokine-driven proliferation and differentiation of human naive, central memory, and effector memory CD4 + T cells. J. Exp. Med., 2001, Vol. 194, no. 12, pp. 1711-1720.Geginat J., Sallusto F., Lanzavecchia A. Cytokine-driven proliferation and differentiation of human naive, central memory, and effector memory CD4 + T cells. J. Exp. Med., 2001, Vol. 194, no. 12, pp. 1711-1720.Greter M., Helft J., Chow A., Hashimoto D., Mortha A., Agudo-Cantero J., Boqunovic M., Gautier E.L., Miller J., Leboeuf M., Lu G., Aloman C., Brown B.D., Pollard J.W., Xiong H., Randolph G.J., Chipuk J.E., Frenette P.S., Merad M. GM-CSF controls nonlymphoid tissue dendritic cell homeostasis but is dispensable for the differentiation of inflammatory dendritic cells. Immunity, 2012, Vol. 36, no. 6, pp. 1031-1046.Greter M., Helft J., Chow A., Hashimoto D., Mortha A., Agudo-Cantero J., Boqunovic M., Gautier E.L., Miller J., Leboeuf M., Lu G., Aloman C., Brown B.D., Pollard J.W., Xiong H., Randolph G.J., Chipuk J.E., Frenette P.S., Merad M. GM-CSF controls nonlymphoid tissue dendritic cell homeostasis but is dispensable for the differentiation of inflammatory dendritic cells. Immunity, 2012, Vol. 36, no. 6, pp. 1031-1046.Halstead, E. S., Umstead, T. M., Davies, M. L., Kawasawa, Y. I., Silveyra, P., Howyrlak, J., Yang L., Guo W., Hu S., Hewage E.K., Chroneos Z.C. GM-CSF overexpression after influenza a virus infection prevents mortality and moderates M1-like airway monocyte/macrophage polarization. Resp. Res., 2018, Vol. 19, no. 1, p. 3.Halstead, E. S., Umstead, T. M., Davies, M. L., Kawasawa, Y. I., Silveyra, P., Howyrlak, J., Yang L., Guo W., Hu S., Hewage E.K., Chroneos Z.C. GM-CSF overexpression after influenza a virus infection prevents mortality and moderates M1-like airway monocyte/macrophage polarization. Resp. Res., 2018, Vol. 19, no. 1, p. 3.Hamilton T.A., Zhao C., Pavicic Jr P. G., Datta S. (2014). Myeloid colony-stimulating factors as regulators of macrophage polarization. Front. Immunol., Vol. 5, p. 554.Hamilton T.A., Zhao C., Pavicic Jr P. G., Datta S. (2014). Myeloid colony-stimulating factors as regulators of macrophage polarization. Front. Immunol., Vol. 5, p. 554.Hercus T.R., Dhagat U., Kan W.L., Broughton S.E., Nero T.L., Perugini M., Sandow J.J., d’Andrea R.J., Ekert P.G., Hughes T., Parker M.W., Lopez A.F. Signalling by the βc family of cytokines. Cytokine Growth Factor Rev., 2013, Vol. 24, no. 3, pp. 189-201.Hercus T.R., Dhagat U., Kan W.L., Broughton S.E., Nero T.L., Perugini M., Sandow J.J., d’Andrea R.J., Ekert P.G., Hughes T., Parker M.W., Lopez A.F. Signalling by the βc family of cytokines. Cytokine Growth Factor Rev., 2013, Vol. 24, no. 3, pp. 189-201.Hercus T.R., Thomas D., Guthridge M.A., Ekert P.G., King-Scott J., Parker M.W., Lopez A.F. The granulocytemacrophage colony-stimulating factor receptor: linking its structure to cell signaling and its role in disease. Blood, 2009, Vol. 114, no. 7, pp. 1289-1298.Hercus T.R., Thomas D., Guthridge M.A., Ekert P.G., King-Scott J., Parker M.W., Lopez A.F. The granulocytemacrophage colony-stimulating factor receptor: linking its structure to cell signaling and its role in disease. Blood, 2009, Vol. 114, no. 7, pp. 1289-1298.Hurdayal R., Brombacher F. Interleukin-4 receptor alpha: from innate to adaptive immunity in murine models of Cutaneous Leishmaniasis. Front. Immunol., 2017, Vol. 8, 1354. doi: 10.3389/fimmu.2017.01354.Hurdayal R., Brombacher F. Interleukin-4 receptor alpha: from innate to adaptive immunity in murine models of Cutaneous Leishmaniasis. Front. Immunol., 2017, Vol. 8, 1354. doi: 10.3389/fimmu.2017.01354.Lau M.Y., Dharmage S.C., Burgess J.A., Win A.K., Lowe A.J., Lodge C., Perret G., Hui J., Thomas P.S., Morrison S., Giles G.G., Hopper J., Abramson M.J., Walters E.H., Matheson M.C. The interaction between farming/ rural environment and TLR2, TLR4, TLR6 and CD14 genetic polymorphisms in relation to early-and late-onset asthma. Sci. Rep., 2017, Vol. 7, 43681. doi: 10.1038/srep43681.Lau M.Y., Dharmage S.C., Burgess J.A., Win A.K., Lowe A.J., Lodge C., Perret G., Hui J., Thomas P.S., Morrison S., Giles G.G., Hopper J., Abramson M.J., Walters E.H., Matheson M.C. The interaction between farming/ rural environment and TLR2, TLR4, TLR6 and CD14 genetic polymorphisms in relation to early-and late-onset asthma. Sci. Rep., 2017, Vol. 7, 43681. doi: 10.1038/srep43681.Mach N., Gillessen S., Wilson S.B., Sheehan C., Mihm M., Dranoff G. Differences in dendritic cells stimulated in vivoby tumors engineered to secrete granulocyte-macrophage colony-stimulating factor or Flt3-ligand. Cancer Res., 2000, Vol. 60, no. 12, pp. 3239-3246.Mach N., Gillessen S., Wilson S.B., Sheehan C., Mihm M., Dranoff G. Differences in dendritic cells stimulated in vivoby tumors engineered to secrete granulocyte-macrophage colony-stimulating factor or Flt3-ligand. Cancer Res., 2000, Vol. 60, no. 12, pp. 3239-3246.Meniailo M.E., Malashchenko V.V., Shmarov V.A., Gazatova N.D., Melashchenko O.B., Goncharov A.G., Seledtsova G.V., Seledtsov V.I. Interleukin-8 favors pro-inflammatory activity of human monocytes/macrophages. Int. Immunopharmacol., 2018, Vol. 56, pp. 217-221.Meniailo M.E., Malashchenko V.V., Shmarov V.A., Gazatova N.D., Melashchenko O.B., Goncharov A.G., Seledtsova G.V., Seledtsov V.I. Interleukin-8 favors pro-inflammatory activity of human monocytes/macrophages. Int. Immunopharmacol., 2018, Vol. 56, pp. 217-221.Mukherjee R., Barman P.K., Thatoi P.K., Tripathy R., Das B.K., Ravindran B. Non-classical monocytes display inflammatory features: validation in sepsis and systemic lupus erythematous. Sci. Rep., 2015, Vol. 5, 13886. doi: 10.1038/srep13886.Mukherjee R., Barman P.K., Thatoi P.K., Tripathy R., Das B.K., Ravindran B. Non-classical monocytes display inflammatory features: validation in sepsis and systemic lupus erythematous. Sci. Rep., 2015, Vol. 5, 13886. doi: 10.1038/srep13886.Seledtsov V.I., Seledtsova G.V. A balance between tissue-destructive and tissue-protective immunities: a role of toll-like receptors in regulation of adaptive immunity. Immunobiology, 2012, Vol. 217, no. 4, pp. 430-435.Seledtsov V.I., Seledtsova G.V. A balance between tissue-destructive and tissue-protective immunities: a role of toll-like receptors in regulation of adaptive immunity. Immunobiology, 2012, Vol. 217, no. 4, pp. 430-435.Shapouri-Moghaddam A., Mohammadian S., Vazini H., Taghadosi M., Esmaeili S.A., Mardani F., Seifi B., Mohammadi A., Afshari J.T., Sahebkar, A. Macrophage plasticity, polarization, and function in health and disease. J. Cell. Physiol., 2018, Vol. 233, no. 9, pp. 6425-6440.Shapouri-Moghaddam A., Mohammadian S., Vazini H., Taghadosi M., Esmaeili S.A., Mardani F., Seifi B., Mohammadi A., Afshari J.T., Sahebkar, A. Macrophage plasticity, polarization, and function in health and disease. J. Cell. Physiol., 2018, Vol. 233, no. 9, pp. 6425-6440.Shiomi A., Usui T. Pivotal roles of GM-CSF in autoimmunity and inflammation. Mediators Inflamm., 2015, Vol. 2015, 568543. doi:10.1155/2015/568543.Shiomi A., Usui T. Pivotal roles of GM-CSF in autoimmunity and inflammation. Mediators Inflamm., 2015, Vol. 2015, 568543. doi:10.1155/2015/568543.Sielska M., Przanowski P., Wylot B., Gabrusiewicz K., Maleszewska M., Kijewska M., Zawadzka M., Kucharska J., Vinnakota K., Kettenmann H., Kotulska K., Grajkowska W., Kaminska B. Distinct roles of CSF family cytokines in macrophage infiltration and activation in glioma progression and injury response. J. Pathol., 2013, Vol. 230, no. 3, pp. 310-321.Sielska M., Przanowski P., Wylot B., Gabrusiewicz K., Maleszewska M., Kijewska M., Zawadzka M., Kucharska J., Vinnakota K., Kettenmann H., Kotulska K., Grajkowska W., Kaminska B. Distinct roles of CSF family cytokines in macrophage infiltration and activation in glioma progression and injury response. J. Pathol., 2013, Vol. 230, no. 3, pp. 310-321.Son Y., Kim B.Y., Park Y.C., Kim K. Diclofenac Inhibits 27-hydroxycholesterol-induced differentiation of monocytic cells into mature dendritic cells. Immune Netw., 2017, Vol. 17, no. 3, pp. 179-185.Son Y., Kim B.Y., Park Y.C., Kim K. Diclofenac Inhibits 27-hydroxycholesterol-induced differentiation of monocytic cells into mature dendritic cells. Immune Netw., 2017, Vol. 17, no. 3, pp. 179-185.van de Laar L., Coffer P.J., Woltman A.M. Regulation of dendritic cell development by GM-CSF: molecular control and implications for immune homeostasis and therapy. Blood, 2012, Vol. 119, no. 15, pp. 3383-3393.van de Laar L., Coffer P.J., Woltman A.M. Regulation of dendritic cell development by GM-CSF: molecular control and implications for immune homeostasis and therapy. Blood, 2012, Vol. 119, no. 15, pp. 3383-3393.Vogel D.Y., Glim J.E., Stavenuiter A.W., Breur M., Heijnen P., Amor S., Dijkstra C.D., Beelen R.H. Human macrophage polarization in vitro:maturation and activation methods compared. Immunobiology, 2014, Vol. 219, no. 9, pp. 695-703.Vogel D.Y., Glim J.E., Stavenuiter A.W., Breur M., Heijnen P., Amor S., Dijkstra C.D., Beelen R.H. Human macrophage polarization in vitro:maturation and activation methods compared. Immunobiology, 2014, Vol. 219, no. 9, pp. 695-703.Wicks I.P., Roberts A.W. Targeting GM-CSF in inflammatory diseases. Nat. Rev. Rheumatol., 2016, Vol. 12. no. 1, pp. 37-48.Wicks I.P., Roberts A.W. Targeting GM-CSF in inflammatory diseases. Nat. Rev. Rheumatol., 2016, Vol. 12. no. 1, pp. 37-48.

The authors declare that there are no conflicts of interest present.