zdorПроблемы здоровья и экологииHealth and Ecology Issues2220-09672708-6011Gomel State Medical University10.51523/2708-6011.2025-22-2-08zdor-2819Research ArticleЭКСПЕРИМЕНТАЛЬНАЯ МЕДИЦИНА И БИОЛОГИЯEXPERIMENTAL MEDICINE AND BIOLOGYИсследование потенциала толерогенных дендритных клеток к реверсии под влиянием фактора некроза опухолиStudy of the potential of tolerogenic dendritic cells to immunophenotypic reversion affected by tumor necrosis factorhttps://orcid.org/0009-0003-2125-4990МиничЯ. С.MinichYa. S.

Минич Яна Сергеевна - научный сотрудник лаборатории иммунологии и вирусологии.

Минск

Yana S. Minich - Researcher at the Laboratory of Immunology and Virology, Institute of Biophysics and Cell Engineering of the National Academy of Sciences of Belarus.

Minsk

yanaminich1094@gmail.comhttps://orcid.org/0000-0002-4869-9864ГончаровА. Е.HancharouA. Y.

Гончаров Андрей Евгеньевич - к.м.н., доцент, директор.

Минск

Andrei Y. Hancharou - Candidate of Medical Sciences, Associate Professor, Director of the Institute of Biophysics and Cell Engineering of the National Academy of Sciences of Belarus.

Minsk

andrei.hancharou@gmail.comhttps://orcid.org/0000-0001-9535-7157АнтоневичН. Г.AntonevichN. G.

Антоневич Наталья Георгиевна - к.б.н., доцент, ведущий научный сотрудник лаборатории иммунологии и вирусологии.

Минск

Natalia G. Antonevich - Candidate of Biological Sciences, Associate Professor, Leading Researcher at the Laboratory of Immunology and Virology, Institute of Biophysics and Cell Engineering of the National Academy of Sciences of Belarus.

Minsk

antonevich.n@gmail.comИнститут биофизики и клеточной инженерии Национальной академии наук БеларусиInstitute of Biophysics and Cell Engineering of National Academy of Sciences of Belarus2025290620252226975Copyright © Минич Я.С., Гончаров А.Е., Антоневич Н.Г., 20252025Минич Я.С., Гончаров А.Е., Антоневич Н.Г.Minich Y.S., Hancharou A.Y., Antonevich N.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.gsmu.by/jour/article/view/2819Цель исследования

Цель исследования. Оценить реверсию толерогенных дендритных клеток (толДК), полученных из человеческих моноцитов крови, в иммуногенные дендритные клетки (ДК) под действием TNF-α in vitro.

Материалы и методы

Материалы и методы. Методом проточной цитофлуориметрии оценивали иммунофенотип ДК, полученных из моноцитов крови здоровых доноров (n = 6), при различных условиях культивирования: культура незрелых ДК, культура зрелых ДК, культура толДК, культура толДК после воздействия провоспалительного фактора TNF-α. Статистическую обработку полученных данных проводили с использованием непараметрических методов.

Результаты

Результаты. В популяции зрелых ДК количество клеток, экспрессирующих молекулу CD83 — 24,05 (19,30–28,20) % достоверно выше, по сравнению с незрелыми ДК, толДК и толДК после стимулирования TNF-α (незрДК — 4,10 (2,75–5,83), толДК — 4,45 (3,68–4,93), толДК/TNF-α — 9,70(8,83–10,58)). Интенсивность экспрессии HLA-DR на поверхности зрелых ДК достоверно выше по сравнению с незрелыми ДК, толДК и толДК после стимулирования TNF-α (незрДК — 23,03(18,24–25,40) усл. ед.; зрДК — 27,54 (25,04–32,41) усл. ед.; толДК — 10,56 (9,21–12,77) усл. ед.; толДК/TNF-α — 13,74 (10,66–18,53) усл. ед.).

Заключение

Заключение. Добавление индуктора созревания TNF-α не приводит к реверсии иммунофенотипа толДК до уровня зрелых ДК. Полученные результаты свидетельствуют в пользу возможности безопасного применения толДК в качестве биомедицинского клеточного продукта для лечения заболеваний, связанных с избыточным иммунным ответом.

Objective

Objective. To assess reversion of tolerogenic dendritic cells (tolDC), obtained from human blood monocytes into immunogenic mature dendritic cells (mDC) under the influence of TNF-α.

Materials and methods

Materials and methods. The immunophenotype of dendritic cells (DCs) obtained from blood monocytes of healthy donors (n=6) was assessed by flow cytometry: immature DC culture (immDC), mature (mDC) culture, tolDC culture, tolDC culture after exposure to the proinflammatory factor TNF-α (tol DCs/TNF-α). Statistical analysis was performed using nonparametric methods.

Results

Results. There was 24.05(19.30-28.20) % of CD83+ cells in the population of mDCs, that was significantly higher compared with immDCs, tolDCs and tolDCs after stimulation with TNF-α (imm DCs – 4.10 (2.75-5.83), tol DCs – 4.45 (3.68-4.93), tol DCs/TNF-α – 9.70 (8.83-10.58). The intensity of HLA-DR expression on the surface of mDCs was significantly higher compared with immDCs, tolDCs and tolDCs after stimulation with TNF-α (imm DCs – 23.03 (18.24-25.40) RFI; tolDCs – 27.54 (25.04-32.41) RFI; tolDC – 10.56 (9.21-12.77) RFI; tolDC/TNF-α – 13.74 (10.66-18.53) RFI (RFI – relative fluorescence intensity).

Conclusion

Conclusion. Adding of the TNF-α as maturation inducer did not result in immunophenotypic reversion of tolDCs to the mDCs level. The obtained data indicate in favor of possibility to use safely tolDCs as a biomedical cell product for the treatment of diseases associated with an excessive immune response.

толерогенные дендритные клеткипроточная цитометрияиммунофенотипированиеtolerogenic dendritic cellsflow cytometryimmunophenotypingИсследование выполнено в рамках мероприятия 23-й Государственной программы «Наукоемкие технологии и техника» 2021–2025 гг. (№ 20213421).This research was carried out within activities of the 23rd State program “High Technologies and Technique” 2021-2025 (No. 20213421).ReferencesAdorini L, Giarratana N, Penna G. Pharmacological induction of tolerogenic dendritic cells and regulatory T cells. Semin Immunol. 2004;16:127-134. DOI: https://doi.org/10.1016/j.smim.2003.12.008Adorini L, Giarratana N, Penna G. Pharmacological induction of tolerogenic dendritic cells and regulatory T cells. Semin Immunol. 2004;16:127-134. DOI: https://doi.org/10.1016/j.smim.2003.12.008Piemonti L, Monti P, Sironi M, Fraticelli P, Leone BE, et al. Vitamin D3 affects differentiation, maturation, and function of human monocyte-derived dendritic cells. J Immunol. 2000;164:4443-4451. DOI: https://doi.org/10.4049/jimmunol.164.9.4443Piemonti L, Monti P, Sironi M, Fraticelli P, Leone BE, et al. Vitamin D3 affects differentiation, maturation, and function of human monocyte-derived dendritic cells. J Immunol. 2000;164:4443-4451. DOI: https://doi.org/10.4049/jimmunol.164.9.4443Svajger U, Rozman P. Induction of Tolerogenic Dendritic Cells by Endogenous Biomolecules: An Update. Frontiers in Immunology. 2018;9. DOI: https://doi.org/10.3389/fimmu.2018.02482Svajger U, Rozman P. Induction of Tolerogenic Dendritic Cells by Endogenous Biomolecules: An Update. Frontiers in Immunology. 2018;9. DOI: https://doi.org/10.3389/fimmu.2018.02482Svajger U, Rozman P. Tolerogenic dendritic cells: molecular and cellular mechanisms in transplantation. J Leukoc Biol. 2014;95:53-69. DOI: https://doi.org/10.1189/jlb.0613336Svajger U, Rozman P. Tolerogenic dendritic cells: molecular and cellular mechanisms in transplantation. J Leukoc Biol. 2014;95:53-69. DOI: https://doi.org/10.1189/jlb.0613336Bell GM, Anderson AE, Diboll J, Reece R, Eltherington O, Harry RA, et al. Autologous tolerogenic dendritic cells for rheumatoid and inflammatory arthritis. Annals of the Rheumatic Diseases. 2017;76(1):227-234. DOI: https://doi.org/10.1136/annrheumdis-2015-208456Bell GM, Anderson AE, Diboll J, Reece R, Eltherington O, Harry RA, et al. Autologous tolerogenic dendritic cells for rheumatoid and inflammatory arthritis. Annals of the Rheumatic Diseases. 2017;76(1):227-234. DOI: https://doi.org/10.1136/annrheumdis-2015-208456Гончаров А.Е., Антоневич Н.Г., Чекан В.Л. Влияние мезенхимальных стволовых клеток обонятельной выстилки на антигенный профиль дендритных клеток. Новости медико-биол. наук. 2015;12(4):159-162.Hancharou A, Antonevich N, Chekan V. The influence of olfactory mucosa-derived mesenchymal stem cells on the antigenic profile of the dendritic cells. News of Biomedical Sciences. 2015;12(4):159-162. (In Russ.).Гончаров А.Е., Минич Я.С., Антоневич Н.Г. Иммуноморфологическая характеристика толерогенных дендритных клеток. Новости медико-биол. наук. 2024;24(3):182-194.Hancharou A, Minich Y, Antonevich N. Immunomorphological characteristics of tolerogenic dendritic cells. News of Biomedical Sciences. 2024;24(3):182-194. (In Russ.).Domogalla MP, Rostan PV, Raker VK, et al. Tolerance through education: how tolerogenic dendritic cells shape immunity. Frontiers in Immunology. 2017;8:1764. DOI: https://doi.org/10.3389/fimmu.2017.01764Domogalla MP, Rostan PV, Raker VK, et al. Tolerance through education: how tolerogenic dendritic cells shape immunity. Frontiers in Immunology. 2017;8:1764. DOI: https://doi.org/10.3389/fimmu.2017.01764Hilkens CMU, Isaacs JD. Tolerogenic dendritic cell therapy for rheumatoid arthritis: where are we now? Clinical & Experimental Immunology. 2013;172(2):148-157. DOI: https://doi.org/10.1111/cei.12038Hilkens CMU, Isaacs JD. Tolerogenic dendritic cell therapy for rheumatoid arthritis: where are we now? Clinical & Experimental Immunology. 2013;172(2):148-157. DOI: https://doi.org/10.1111/cei.12038Raker VK, Domogalla MP, Steinbrink K. Tolerogenic Dendritic Cells for Regulatory T Cell Induction in Man. Frontiers in Immunology. 2015;6:569. DOI: https://doi.org/10.3389/fimmu.2015.00569Raker VK, Domogalla MP, Steinbrink K. Tolerogenic Dendritic Cells for Regulatory T Cell Induction in Man. Frontiers in Immunology. 2015;6:569. DOI: https://doi.org/10.3389/fimmu.2015.00569Wu L, Liu YJ. Development of dendritic-cell lineages. Immunity. 2007;26(6):741-750. DOI: https://doi.org/10.1016/j.immuni.2007.06.006Wu L, Liu YJ. Development of dendritic-cell lineages. Immunity. 2007;26(6):741-750. DOI: https://doi.org/10.1016/j.immuni.2007.06.006Blanco P, Palucka AK, Pascual V, Banchereau J. Dendritic cells and cytokines in human inflammatory and autoimmune diseases. Cytokine Growth Factor Rev. 2008;19(1):41-52. DOI: https://doi.org/10.1016/j.cytogfr.2007.10.004Blanco P, Palucka AK, Pascual V, Banchereau J. Dendritic cells and cytokines in human inflammatory and autoimmune diseases. Cytokine Growth Factor Rev. 2008;19(1):41-52. DOI: https://doi.org/10.1016/j.cytogfr.2007.10.004Trevejo JM, Marino MW, Philpott N, Josien R, et al. TNF-alpha -dependent maturation of local dendritic cells is critical for activating the adaptive immune response to virus infection. Proc Natl Acad Sci USA. 2001;98(21):12162-12167. DOI: https://doi.org/10.1073/pnas.211423598Trevejo JM, Marino MW, Philpott N, Josien R, et al. TNF-alpha -dependent maturation of local dendritic cells is critical for activating the adaptive immune response to virus infection. Proc Natl Acad Sci USA. 2001;98(21):12162-12167. DOI: https://doi.org/10.1073/pnas.211423598Ritter U, Meissner A, Ott J, Körner H. Analysis of the maturation process of dendritic cells deficient for TNF and lymphotoxin-alpha reveals an essential role for TNF. J Leukoc Biol. 2003;74:216-222. DOI: https://doi.org/10.1189/jlb.1202587Ritter U, Meissner A, Ott J, Körner H. Analysis of the maturation process of dendritic cells deficient for TNF and lymphotoxin-alpha reveals an essential role for TNF. J Leukoc Biol. 2003;74:216-222. DOI: https://doi.org/10.1189/jlb.1202587Тимохина О.В., Гончаров А.Е., Антоневич Н.Г. Методы получения дендритных клеток для лечения онкологических заболеваний. Новости медико-биологических наук. 2020;20(2):127-136.Timohina O, Hancharou A, Antonevich N. Methods for obtaining dendritic cells for the treatment of cancer. News Of Biomedical Sciences. 2020;20(2):127-136. (In Russ.).Riaz B, Islam SMS, Ryu HM, Sohn S. Regulates the Immune Responses in Inflammatory Disorders. Int J Mol Sci. 2023;24(3):2831. DOI: https://doi.org/10.3390/ijms24032831Riaz B, Islam SMS, Ryu HM, Sohn S. Regulates the Immune Responses in Inflammatory Disorders. Int J Mol Sci. 2023;24(3):2831. DOI: https://doi.org/10.3390/ijms24032831

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