zdorПроблемы здоровья и экологииHealth and Ecology Issues2220-09672708-6011Gomel State Medical University10.51523/2708-6011.2024-21-2-12zdor-2718Research ArticleЭКСПЕРИМЕНТАЛЬНАЯ МЕДИЦИНА И БИОЛОГИЯEXPERIMENTAL MEDICINE AND BIOLOGYВлияние пулирования мезенхимальных стволовых клеток на подавление пролиферативной активности Т-лимфоцитовInfluence of pooling mesenchymal stem cells on the suppression of proliferative activity of T-lymphocyteshttps://orcid.org/0009-0001-2218-2349РындаЕ. Г.RyndaA. H.

Рында Елена Геннадьевна, аспирант лаборатории иммунологии и вирусологии

г. Минск

 

Alena H. Rynda, Postgraduate Student at the Laboratory of Immunology and Virology

Minsk

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

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

г. Минск

Andrei Y. Hancharou, Candidate of Medical Sciences, Associate Professor, Director 

Minsk

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

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

г. Минск

Natalia G. Antonevich, Candidate of Biological Sciences, Leading Researcher

Minsk

antonevich.n@gmail.comИнститут биофизики и клеточной инженерии Национальной академии наук БеларусиInstitute of Biophysics and Cell Engineering of the National Academy of Sciences of Belarus20242705202421297102Copyright © Рында Е.Г., Гончаров А.Е., Антоневич Н.Г., 20242024Рында Е.Г., Гончаров А.Е., Антоневич Н.Г.Rynda A.H., 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/2718Цель исследования

Цель исследования. Определить влияние пулирования (объединения) одиночных культур мезенхимальных стволовых клеток обонятельной выстилки (МСК ОВ) носовой полости на подавление пролиферативной активности Т-лимфоцитов.

Материалы и методы

Материалы и методы. Методом проточной цитометрии была проведена оценка влияния пулирования одиночных культур МСК ОВ, полученных от здоровых доноров (n = 7), на митоген-индуцированную пролиферацию Т-лимфоцитов периферической крови добровольцев (n = 5). Оценку индекса пролиферации и числа поделившихся CD3+ клеток проводили по изменению интенсивности флуоресценции красителя Tag-it Vio. Статистическую обработку полученных данных проводили с использованием непараметрических методов.

Результаты

Результаты. С учетом использования пяти образцов периферической крови добровольцев выполнена оценка 35 вариантов сокультивирования одиночных МСК с Т-клетками и 33 вариантов сокультивирования пулМСК с Т-клетками. Был установлен ингибирующий эффект как одиночных (р = 0,0001), так и пулМСК ОВ (р = 0,0001) на ФГА-индуцированную пролиферацию CD3+ Т-клеток суммарной фракции лимфоцитов периферической крови. Показано, что пулМСК ОВ оказывают достоверно более выраженный супрессивный эффект на пролиферацию Т-клеток в сравнении с одиночными МСК (р = 0,000004).

Заключение

Заключение. ПулМСК оказывают достоверно более выраженный эффект в отношении подавления ФГА-индуцированной пролиферации Т-клеток периферической крови в сравнении с монокультурами МСК, что обосновывает применение пулМСК в медицине с целью достижения лучших результатов в лечении иммуноопосредованных заболеваний.

Objective

Objective. To determine the effect of pooling (merging) of single cultures of olfactory mucosa-derived mesenchymal stem cells (MSCs) of the nasal cavity on the suppression of the proliferative activity of T-lymphocytes.

Materials and methods

Materials and methods. Using flow cytometry, the effect of pooling single cultures of MSCs obtained from healthy donors (n=7) on the mitogen-induced proliferation of T-lymphocytes in the peripheral blood of volunteers (n=5) was studied. The proliferation index and the number of dividing CD3+ T-cells were assessed by changes in the fluorescence intensity of Tag-it Vio. Statistical processing of the obtained data was carried out using non-parametric statistics.

Results

Results. An evaluation of 35 variants of co-culture of single MSCs with T-cells and 33 variants of co-culture of pooled MSCs with T-cells was performed. The inhibitory effect of both single (p = 0.0001) and pooled MSCs (p = 0.0001) on

PHA-induced proliferation of CD3+ T-cells in the total fraction of peripheral blood lymphocytes was identified. It was shown that pooled MSCs have a significantly more pronounced suppressive effect on T-cell proliferation compared to single MSCs (p=0.000004).

Conclusion

Conclusion. PoolMSCs have a significantly more pronounced effect in suppressing PHA-induced proliferation of peripheral blood T-cells compared to MSC monocultures, which justifies the use of poolMSCs in medicine to achieve better results in the treatment of immune-mediated diseases.

пулированные мезенхимальные стволовые клеткипроточная цитометрияиммунофенотипированиестволовые клеткиpooled mesenchymal stem cellsflow cytometryimmunophenotypingstem cellsReferencesRodríguez-Fuentes DE, Fernández-Garza LE, Samia-Meza JA, Barrera-Barrera SA, Caplan AI, Barrera-Saldaña HA. Mesenchymal stem cells current clinical applications: a systematic review. Archives of Medical Research. 2021;52(1):93-101. DOI: https://doi.org/10.1016/j.arcmed.2020.08.006Rodríguez-Fuentes DE, Fernández-Garza LE, Samia-Meza JA, Barrera-Barrera SA, Caplan AI, Barrera-Saldaña HA. Mesenchymal stem cells current clinical applications: a systematic review. Archives of Medical Research. 2021;52(1):93-101. DOI: https://doi.org/10.1016/j.arcmed.2020.08.006Pittenger MF, Discher DE, Péault BM, Phinney DG, Hare JM, Caplan AI. Mesenchymal stem cell perspective: cell biology to clinical progress. NPJ Regenerative Medicine. 2019;4(1):22. DOI: https://doi.org/10.1038/s41536-019-0083-6Pittenger MF, Discher DE, Péault BM, Phinney DG, Hare JM, Caplan AI. Mesenchymal stem cell perspective: cell biology to clinical progress. NPJ Regenerative Medicine. 2019;4(1):22. DOI: https://doi.org/10.1038/s41536-019-0083-6Jiang W, Xu J. Immune modulation by mesenchymal stem cells. Cell Proliferation. 2020;53(1): e12712. DOI: https://doi.org/10.1111/cpr.12712Jiang W, Xu J. Immune modulation by mesenchymal stem cells. Cell Proliferation. 2020;53(1): e12712. DOI: https://doi.org/10.1111/cpr.12712Huang Y, Yin Y, Gu Y, Gu Q, Yang H, Zhou Z, et al. Characterization and immunogenicity of bone marrow-derived mesenchymal stem cells under osteoporotic conditions. Science China Life Sciences. 2020;63:429-442. DOI: https://doi.org/10.1007/s11427-019-1555-9Huang Y, Yin Y, Gu Y, Gu Q, Yang H, Zhou Z, et al. Characterization and immunogenicity of bone marrow-derived mesenchymal stem cells under osteoporotic conditions. Science China Life Sciences. 2020;63:429-442. DOI: https://doi.org/10.1007/s11427-019-1555-9Wang Y, Tian M, Wang F, Heng BC, Zhou J, Cai Z, et al. Understanding the immunological mechanisms of mesenchymal stem cells in allogeneic transplantation: from the aspect of major histocompatibility complex class I. Stem Cells and Development. 2019;28(17):1141-1150. DOI: https://doi.org/10.1089/scd.2018.0256Wang Y, Tian M, Wang F, Heng BC, Zhou J, Cai Z, et al. Understanding the immunological mechanisms of mesenchymal stem cells in allogeneic transplantation: from the aspect of major histocompatibility complex class I. Stem Cells and Development. 2019;28(17):1141-1150. DOI: https://doi.org/10.1089/scd.2018.0256Ganguly, P., El-Jawhari, J. J., Giannoudis, P. V., Burska, A. N., Ponchel, F., Jones, E. A. (2017). Age-related changes in bone marrow mesenchymal stromal cells: a potential impact on osteoporosis and osteoarthritis development. Cell transplantation. 2017; 26(9): 1520-1529.Ganguly, P., El-Jawhari, J. J., Giannoudis, P. V., Burska, A. N., Ponchel, F., Jones, E. A. (2017). Age-related changes in bone marrow mesenchymal stromal cells: a potential impact on osteoporosis and osteoarthritis development. Cell transplantation. 2017; 26(9): 1520-1529.Wang, Q., Xu, N., Wang, Y., Zhang, X., Liu, L., Zhou, H. et al. Allogeneic stem cell transplantation combined with transfusion of mesenchymal stem cells in primary myelofibrosis: a multicenter retrospective study. Frontiers in Oncology. 2022; 11: 792142. DOI: https://doi.org/10.3389/fonc.2021.792142Wang, Q., Xu, N., Wang, Y., Zhang, X., Liu, L., Zhou, H. et al. Allogeneic stem cell transplantation combined with transfusion of mesenchymal stem cells in primary myelofibrosis: a multicenter retrospective study. Frontiers in Oncology. 2022; 11: 792142. DOI: https://doi.org/10.3389/fonc.2021.792142Fijany A, Sayadi LR, Khoshab N, Banyard DA, Shaterian A., Alexander M, et al. Mesenchymal stem cell dysfunction in diabetes. Molecular Biology Reports. 2019;46:1459-1475. DOI: https://doi.org/10.1007/s11033-018-4516-xFijany A, Sayadi LR, Khoshab N, Banyard DA, Shaterian A., Alexander M, et al. Mesenchymal stem cell dysfunction in diabetes. Molecular Biology Reports. 2019;46:1459-1475. DOI: https://doi.org/10.1007/s11033-018-4516-xPedrosa M, Gomes J, Laranjeira P, Duarte C, Pedreiro S, Antunes B, et al. Immunomodulatory effect of human bone marrow‐derived mesenchymal stromal/stem cells on peripheral blood T cells from rheumatoid arthritis patients. Journal of Tissue Engineering and Regenerative Medicine. 2020;14(1):16-28.Pedrosa M, Gomes J, Laranjeira P, Duarte C, Pedreiro S, Antunes B, et al. Immunomodulatory effect of human bone marrow‐derived mesenchymal stromal/stem cells on peripheral blood T cells from rheumatoid arthritis patients. Journal of Tissue Engineering and Regenerative Medicine. 2020;14(1):16-28.Cheng RJ, Xiong AJ, Li YH, Pan SY, Zhang QP, Zhao Y, et al. Mesenchymal stem cells: allogeneic MSC may be immunosuppressive but autologous MSC are dysfunctional in lupus patients. Frontiers in Cell and Developmental Biology. 2019;7:285. DOI: https://doi.org/10.3389/fcell.2019.00285Cheng RJ, Xiong AJ, Li YH, Pan SY, Zhang QP, Zhao Y, et al. Mesenchymal stem cells: allogeneic MSC may be immunosuppressive but autologous MSC are dysfunctional in lupus patients. Frontiers in Cell and Developmental Biology. 2019;7:285. DOI: https://doi.org/10.3389/fcell.2019.00285Jiang W, Xu J. Immune modulation by mesenchymal stem cells. Cell Proliferation. 2020;53(1):e12712. DOI: https://doi.org/10.1111/cpr.12712Jiang W, Xu J. Immune modulation by mesenchymal stem cells. Cell Proliferation. 2020;53(1):e12712. DOI: https://doi.org/10.1111/cpr.12712El-Sayed M, El-Feky MA, El-Amir MI, Hasan AS, Tag-Adeen M, Urata Y, et al. Immunomodulatory effect of mesenchymal stem cells: Cell origin and cell quality variations. Molecular Biology eports. 2019;46:1157-1165. DOI: https://doi.org/10.3389/fimmu.2021.683003El-Sayed M, El-Feky MA, El-Amir MI, Hasan AS, Tag-Adeen M, Urata Y, et al. Immunomodulatory effect of mesenchymal stem cells: Cell origin and cell quality variations. Molecular Biology eports. 2019;46:1157-1165. DOI: https://doi.org/10.3389/fimmu.2021.683003Castro LL, Lopes-Pacheco M, Weiss DJ, Cruz FF, Rocco PRM. Current understanding of the immunosuppressive properties of mesenchymal stromal cells. Journal of Molecular Medicine. 2019;97:605-618.Castro LL, Lopes-Pacheco M, Weiss DJ, Cruz FF, Rocco PRM. Current understanding of the immunosuppressive properties of mesenchymal stromal cells. Journal of Molecular Medicine. 2019;97:605-618.Negi N, Griffin MD. Effects of mesenchymal stromal cells on regulatory T cells: Current understanding and clinical relevance. Stem Cells. 2020;38(5):596-605. DOI: https://doi.org/10.1002/stem.3151Negi N, Griffin MD. Effects of mesenchymal stromal cells on regulatory T cells: Current understanding and clinical relevance. Stem Cells. 2020;38(5):596-605. DOI: https://doi.org/10.1002/stem.3151Kannan S, Viswanathan P, Gupta PK, Kolkundkar UK. Characteristics of pooled wharton’s jelly mesenchymal stromal cells (WJ-MSCs) and their potential role in rheumatoid arthritis treatment. Stem Cell Reviews and Reports. 2022;18(5):1851-1864. DOI: https://doi.org/10.1007/s12015-022-10344-wKannan S, Viswanathan P, Gupta PK, Kolkundkar UK. Characteristics of pooled wharton’s jelly mesenchymal stromal cells (WJ-MSCs) and their potential role in rheumatoid arthritis treatment. Stem Cell Reviews and Reports. 2022;18(5):1851-1864. DOI: https://doi.org/10.1007/s12015-022-10344-wWidholz B, Tsitlakidis S, Reible B, Moghaddam A, Westhauser F. Pooling of patient-derived mesenchymal stromal cells reduces inter-individual confounder-associated variation without negative impact on cell viability, proliferation and osteogenic differentiation. Cells. 2019;8(6):633. DOI: https://doi.org/10.3390/cells8060633Widholz B, Tsitlakidis S, Reible B, Moghaddam A, Westhauser F. Pooling of patient-derived mesenchymal stromal cells reduces inter-individual confounder-associated variation without negative impact on cell viability, proliferation and osteogenic differentiation. Cells. 2019;8(6):633. DOI: https://doi.org/10.3390/cells8060633Kuçi Z, Bönig H, Kreyenberg H, Bunos M, Jauch A, Janssen JW, et al. Mesenchymal stromal cells from pooled mononuclear cells of multiple bone marrow donors as rescue therapy in pediatric severe steroid-refractory graft-versus-host disease: a multicenter survey. Haematologica. 2016;101(8):985. DOI: https://doi.org/10.3324/haematol.2015.140368Kuçi Z, Bönig H, Kreyenberg H, Bunos M, Jauch A, Janssen JW, et al. Mesenchymal stromal cells from pooled mononuclear cells of multiple bone marrow donors as rescue therapy in pediatric severe steroid-refractory graft-versus-host disease: a multicenter survey. Haematologica. 2016;101(8):985. DOI: https://doi.org/10.3324/haematol.2015.140368Hejretová, L., Čedíková, M., Dolejšová, M., Vlas, T., Jindra, P., Lysák, D. et al. Comparison of the immunomodulatory effect of single MSC batches versus pooled MSC products. Cell and Tissue Banking. 2020; 21:119-129.Hejretová, L., Čedíková, M., Dolejšová, M., Vlas, T., Jindra, P., Lysák, D. et al. Comparison of the immunomodulatory effect of single MSC batches versus pooled MSC products. Cell and Tissue Banking. 2020; 21:119-129.

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