MSC-like cells increase ability of monocyte-derived dendritic cells to polarize IL-17-/IL-10-producing T cells via CTLA-4

•Mesenchymal stromal cell-like cells alter moDC differentiation via RARα activation•Mesenchymal express genes known to play role in ATRA synthesis•MoDCs, differentiated the presence of MSCl-derived factors, CTLA-4•CTLA-4+ moDCs are able induce polarization IL-10- and IL-17-producing helper T Mesenchymal (MSCl) generated from human embryonic stem considered be an eligible cell line model immunomodulatory behavior mesenchymal (MSCs) vitro. Dendritic (DCs) essential players maintenance restoration sensitive balance between tolerance immunity. Here, effects MSCl on vitro monocytes into DCs were investigated. promote CTLA-4 expressing production all-trans retinoic acid (ATRA) functioning as a ligand RARα, key nuclear receptor DC development. These semi-matured exhibit ability activate allogeneic, naive polarize them IL-10 + IL-17 double-positive CTLA-4-dependent manner. Mapping molecular mechanisms MSC-mediated indirect modulation may help expand MSCs' clinical application cell-free therapies. Due their multipotent strong potential, promising candidates for cell-based therapy several inflammatory, immune-mediated, degenerative diseases (Saeedi et al., 2019Saeedi P. Halabian R. Imani Fooladi A.A. A revealing review therapy, perspectives Modification strategies.Stem Cell Invest. 2019; 6: 34Crossref PubMed Google Scholar). There number ongoing trials related MSCs or linked graft enhancement utilizing immunosuppressive functions (Wang 2018Wang M. Yuan Q. Xie L. immunomodulation: properties application.Stem Cells Int. 2018; 2018: 3057624Crossref Scopus (227) However, exact cellular underlying immunomodulation have yet revealed. Despite large studies focusing changes tissues following administration, pieces puzzle still missing results often inconsistent. Possible reasons include that different sources under culture conditions diverse phenotype epigenetic background, secrete distinct patterns soluble factors possess capacities proliferate (Weiss Dahlke, 2019Weiss A.R.R. Dahlke M.H. Immunomodulation by (MSCs): action living, apoptotic, dead MSCs.Front. Immunol. 10: 1191Crossref (198) One strategies overcome these limitations is use MSC-like lines derived pluripotent cells, generating unlimited numbers early passage with consistent quality immune suppressive (Kimbrel 2014Kimbrel E.A. Kouris N.A. Yavanian G.J. Chu J. Qin Y. Chan A. Singh R.P. Mccurdy D. Gordon Levinson R.D. Lanza population displays potent therapeutic properties.Stem Dev. 2014; 23: 1611-1624Crossref (115) We earlier demonstrated collaborative study based phenotypic functional such potential activity, HUES9 bone marrow-derived (Varga 2011Varga N. Vereb Z. Rajnavolgyi E. Nemet K. Uher F. Sarkadi B. Apati like support growth.Biochem. Biophys. Res. Commun. 2011; 414: 474-480Crossref (19) In recent study, we provided body evidence modulate responses monocyte-derived dendritic (moDCs) acid-inducible gene I receptor-mediated activation (Bacskai 2015Bacskai I. Mazlo Kis-Toth Szabo Panyi G. set stimulatory inhibitory signals cells.Stem 2015; 24: 1805-1816Crossref (4) Human stimulated exhibited reduced expression indicators activation, lower TNF-α, CXCL10, IL-12 IFNγ, well decreased migration compared activated absence prior observations indicate mature can efficiently suppressed cell-derived mechanisms. this investigated whether how altered cells. To analyze direct MSCl-CM relative cytokine, chemokine other protein levels supernatants using array (Figure S1). It was observed released remarkably secretion profile moDCs. Elevated BAFF, complement factor D, EGF, IL-2, IL-22, lipocalin-2, TGFα, TIM3, myeloperoxidase, C-reactive protein, Dkk-1, while productions ICAM-1 EMMPRIN detected supernatant 1A). increased Vitamin D BP, Endoglin, ENA78, GDF-15, GRO-α, IL-24, MCP-3, VEGF, IL-8, IL-10, but amounts FGF-19, Osteopontin, CD31, IL-18 Bpa found moDC-MSCl co-cultures Both exposure contact raised IL-19, VCAM-1, leptin, IL-6, CD14, FGF basic, IGFBP2, TFF3 TfR, IL-27, Cystatin C, chitinase 3-like 1, MMP-9, PDGF-AB/BB diminished either validate results, concentrations some inflammatory (IL-6) anti-inflammatory (IL-10 TGFβ) mediators determined ELISA 1B). significantly enhanced IL-6 detected, secreted more TGFβ than control demonstrate baseline both cultures modulated through membrane-bound molecules. get insight affects characteristics vitro, monitored surface molecules flow cytometry. During GM-CSF IL-4, CD14 downregulated (Sallusto Lanzavecchia, 1994Sallusto Lanzavecchia Efficient presentation antigen cultured maintained granulocyte/macrophage colony-stimulating plus interleukin 4 tumor necrosis alpha.J. Exp. Med. 1994; 179: 1109-1118Crossref (4433) Scholar), DC-SIGN (Geijtenbeek 2000Geijtenbeek T.B. Torensma Van Vliet S.J. Duijnhoven G.C. Adema Kooyk Figdor C.G. Identification DC-SIGN, novel cell-specific ICAM-3 supports primary responses.Cell. 2000; 100: 575-585Abstract Full Text PDF (1407) Scholar) group 1 CD1 family members upregulated. our experiments, percentage CD14+ higher 2A). While ratio DC-SIGN+ similar treated cultures, molecule MSCl-exposed experimental conditions, approx. 40% untreated expressed including CD1a, CD1b, CD1c 2B). Exposure glycolipid receptors treatments did not modify viability 2C). This observation suggests lipid antigens proteins negatively For further characterization MSCl-CM- cell-treated CD163, macrophage scavenger (Skytthe 2020Skytthe M.K. Graversen J.H. Moestrup S.K. Targeting CD163(+) macrophages malignant diseases.Int. Mol. Sci. 2020; 21: 5497Crossref (25) Interestingly, treatment induced only slight increase frequency CD163+ whereas triggered significant rise marker-expressing S2A). also peptide antigen-presenting HLA-DQ co-stimulatory CD86, CD80 CD86 3A). Augmented PD-L1, co-inhibitory molecule, could 3B). Importantly, another exclusively time-dependent manner statistically difference day 3C, left panel). four-fold at mRNA level four-day-old right When immature co-cultured 48 hr remarkable CD80, PD-L1 S3). when same period time no minor suggesting regulatory stage development together microscopic morphology S2B) has regulate toward CD1a−DC-SIGN+CD163low moDCs, CD1a−DC-SIGNlowCD163high M2 macrophage-like recently been reported specialized blood (Hashimoto-Hill 2018Hashimoto-Hill S. Friesen Park Im Kaplan Kim C.H. RARalpha Langerhans langerin-expressing conventional cells.Nat. 9: 3896Crossref (10) Therefore, investigate guiding MSCl-CM, analyzed process agonist BMS614 selective antagonist receptor. The CD1a lowered (Figures 2C 4A ) 4A). As expected, prevented effect expression. addition insignificant Inhibition blocked intensify presentation; however, differences non-significant largely dependent Indeed, upregulation function inhibited. Furthermore, CTLA-4+ blockade Next, it production. First, synthesis retinol dehydrogenase 10 (RDH10), aldehyde ALDH1A1 ALDH1A3, ALDH1A2 4B). next series activity isoenzymes inhibited highly inhibitor DEAB (Morgan 2015Morgan C.A. Parajuli Buchman C.D. Dria Hurley T.D. N,N-diethylaminobenzaldehyde (DEAB) substrate mechanism-based ALDH isoenzymes.Chem. Biol. Interact. 234: 18-28Crossref (70) deficient monitored. collected enhance 4C). contrast HLA-DQ, exposed Taken together, suggest bring about least partially dissect capacity ELISPOT assays appropriate monitor moDC-mediated allogeneic T-lymphocyte used. producing 5A) 5B). Moreover, cytometric analysis revealed IL-10+IL-17+ 5C). polarization, CTLA-4-mediated signaling pathway specific antibodies. Unexpectedly, MSCl-CM-treated trigger IL-17- IL-10-producing 5A–5C). isotype antibodies affect primed S4). parallel Th1- Th2-polarizing MSCl-CM-exposed S5). (BMS614), which induction 4A), they displayed capability S6). 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