Generation of human myeloid suppressor cells in the in vitro experimental model
- 作者: Timganova V.1, Bochkova M.1, Uzhviyuk S.2, Shardina K.1, Zamorina S.1,2, Rayev M.1,2
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隶属关系:
- Institute of Ecology and Genetics of Microorganisms, Ural Branch, Russian Academy of Sciences, Branch of the Perm Federal Research Center, Ural Branch, Russian Academy of Sciences
- Perm State National Research University
- 期: 卷 23, 编号 2 (2020)
- 页面: 157-162
- 栏目: SHORT COMMUNICATIONS
- URL: https://journals.rcsi.science/1028-7221/article/view/119449
- DOI: https://doi.org/10.46235/1028-7221-352-GOH
- ID: 119449
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Myeloid suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells that generally differentiate into macrophages, granulocytes, and dendritic cells. However, in pathology, these cells acquire a suppressor phenotype, blocking immune response. In particular, MDSC levels increase in many pathological conditions, including inflammation, sepsis, traumatic shock, autoimmune diseases, cancer, and pregnancy. Over the past 12 years, an interest in examining this cell population has been steadily increased [PUBMED: 2008 (65 articles); 2020 (> 650 entries)] that will expand our understanding of immune system functioning. In humans, MDSCs are characterized by HLA-DR-CD33+CD11b+ phenotype, in turn being subdivided into CD15+ or CD66+ granulocytic (G-MDSC), CD14+ monocytic (M-MDSC), and early (e-MDSC) MDSC bearing HLA-DR-CD11b+CD33+CD14-CD66b- phenotype. This work was aimed to develop an adequate experimental model allowing to evaluate cytokine-driven differentiation of human MDSCs from peripheral blood mononuclear cells in long-term in vitro culture system. For this, peripheral blood mononuclear cells were isolated from healthy donors induced to express MDSC phenotype with GM-CSF and IL-6 (40 or 20 ng/ml) cultured for 7, 14, 21 days. In several experiments, LPS (100 ng/ml) was added to the cultured cells 24 hours before immunophenotyping. The percentage of living Zombie Aquanegative cells in cultures (gated on cells according to FSC/SSC) ranged from 90.5-93.9%. No significant differences were observed between cultured cells. In our experimental conditions, the mean percentage of total MDSC subpopulation reached 2-2.3% of total living cells, exceeding that one by 9-10-fold found in freshly isolated mononuclear cells from healthy subjects. Based on the results of our experimental study, we found that induction of e-MDSC derived from human peripheral blood mononuclear cells requires two weeks of co-culture with 40 ng/ml IL-6 and 40 ng/ml GM-CSF. “Mature” MDSCs (M-MDSC + G-MDSC) yield was peaked in the following conditions: co-culture for 3 weeks with 20 ng/ml IL-6 and 20 ng/ml GM-CSF added with 100 ng/ml LPS 24 hours before completing protocol. Overall, further examining factors modulating MDSC differentiation will reveal conditions necessary for generating this suppressor cell subset potentially used in clinical practice.
作者简介
V. Timganova
Institute of Ecology and Genetics of Microorganisms, Ural Branch, Russian Academy of Sciences, Branch of the Perm Federal Research Center, Ural Branch, Russian Academy of Sciences
编辑信件的主要联系方式.
Email: timganovavp@gmail.com
Timganova Valeriya P. - PhD (Biology), Research Associate, Laboratory of Ecological Immunology
614081, Perm, Golev str., 13
Phone: 7 (902) 836-14-55
M. Bochkova
Institute of Ecology and Genetics of Microorganisms, Ural Branch, Russian Academy of Sciences, Branch of the Perm Federal Research Center, Ural Branch, Russian Academy of Sciences
Email: fake@neicon.ru
PhD (Biology), Research Associate, Laboratory of Ecological Immunology
Perm
俄罗斯联邦S. Uzhviyuk
Perm State National Research University
Email: fake@neicon.ru
Student, Microbiology and Immunology Department, Faculty of Biology
Perm
俄罗斯联邦K. Shardina
Institute of Ecology and Genetics of Microorganisms, Ural Branch, Russian Academy of Sciences, Branch of the Perm Federal Research Center, Ural Branch, Russian Academy of Sciences
Email: fake@neicon.ru
Graduate Student, Laboratory of Ecological Immunology
Perm
俄罗斯联邦S. Zamorina
Institute of Ecology and Genetics of Microorganisms, Ural Branch, Russian Academy of Sciences, Branch of the Perm Federal Research Center, Ural Branch, Russian Academy of Sciences; Perm State National Research University
Email: fake@neicon.ru
PhD, MD (Biology), Leading Research Associate, Laboratory of Ecological Immunology; Professor, Microbiology and Immunology Department, Faculty of Biology
Perm
俄罗斯联邦M. Rayev
Institute of Ecology and Genetics of Microorganisms, Ural Branch, Russian Academy of Sciences, Branch of the Perm Federal Research Center, Ural Branch, Russian Academy of Sciences; Perm State National Research University
Email: fake@neicon.ru
PhD, MD (Biology), Leading Research Associate, Laboratory of Ecological Immunology; Professor, Microbiology and Immunology Department, Faculty of Biology
Perm
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