hUCMS hold immunomodulatory properties due to both, production of humoral factors (TGFb, IDO, NO, IL6, PGE2, HGF, VEGF) and expression of HLA-E-F and G molecules that are involved in the tolerogenic process occurring at the fetal-maternal interface (HLA-G being recently deemed to induce Treg cells activation). However, graft of free hUCMS is adversely affected by technical and safety issues. These could be overcome by embodying the cells in a biohybrid system made of alginate-based CpS that provide both, immunoprotection and biochemical exchange. We then aimed to determine whether hUCMS in CpS would contrast pathogenic immune cells that are involved in selective b-cell killing in T1D. PBMCs isolated from 15 patients with recent onset T1D and stained with CFSE, were co-incubated with hUCMS in CpS. Cell phenotypic analysis to either identify Tc subsets, intracellular IFNg, IL17 and FoxP3 by FACS, or measure IL17, IL2, IL10, TNFa,IL6,TGFb in culture supernatants, and additionally, RT-PCR and qPCR on mRNA extracts from cultured cells, immunocytochemistry and western blotting, were performed. We observed that hUCMS in CpS : 1) inhibited Tc proliferation in both T1D and control (CD3+ gated cells); 2) rebalanced Treg/effector ratio in T1D PBMCs (significant decline of IFNg expressing CD4+Th1 cells and increase of CD4+FoxP3+ regulatory cells, IL10 concentration increase in supernatant); 3) showed upregulation of IL6, IL10, FoxP3, AhR, TGFb1 and IL21upon qPCR analysis of mRNA expressed by PBMC in hUCMS in CpS co-culture. sHLA-G5 was significantly upregulated. In summary, we preliminarily showed that hUCMS in CpS reduced pathogenic Tc subsets and potentiated the regulatory counterpart in T1D PBMCs, with a central role played by HLA-G5 in maintainining immunomodulation. . In conclusion, hUCMS in CpS may represent a functional biohybrid artificial delivery system, where molecular products induce efficient immunomodulation in T1D.
Human Umbilical Cord Wharton Jelly-derived Adult Mesenchymal Stem Cells (hUCMS) in Microcapsules (CpS): A Novel Biohybrid System for Immunomodulation in T1D
CALAFIORE, Riccardo;MONTANUCCI, Pia;PESCARA, TERESA;PENNONI, ILARIA;ALUNNO, ALESSIA;GERLI, Roberto
2015
Abstract
hUCMS hold immunomodulatory properties due to both, production of humoral factors (TGFb, IDO, NO, IL6, PGE2, HGF, VEGF) and expression of HLA-E-F and G molecules that are involved in the tolerogenic process occurring at the fetal-maternal interface (HLA-G being recently deemed to induce Treg cells activation). However, graft of free hUCMS is adversely affected by technical and safety issues. These could be overcome by embodying the cells in a biohybrid system made of alginate-based CpS that provide both, immunoprotection and biochemical exchange. We then aimed to determine whether hUCMS in CpS would contrast pathogenic immune cells that are involved in selective b-cell killing in T1D. PBMCs isolated from 15 patients with recent onset T1D and stained with CFSE, were co-incubated with hUCMS in CpS. Cell phenotypic analysis to either identify Tc subsets, intracellular IFNg, IL17 and FoxP3 by FACS, or measure IL17, IL2, IL10, TNFa,IL6,TGFb in culture supernatants, and additionally, RT-PCR and qPCR on mRNA extracts from cultured cells, immunocytochemistry and western blotting, were performed. We observed that hUCMS in CpS : 1) inhibited Tc proliferation in both T1D and control (CD3+ gated cells); 2) rebalanced Treg/effector ratio in T1D PBMCs (significant decline of IFNg expressing CD4+Th1 cells and increase of CD4+FoxP3+ regulatory cells, IL10 concentration increase in supernatant); 3) showed upregulation of IL6, IL10, FoxP3, AhR, TGFb1 and IL21upon qPCR analysis of mRNA expressed by PBMC in hUCMS in CpS co-culture. sHLA-G5 was significantly upregulated. In summary, we preliminarily showed that hUCMS in CpS reduced pathogenic Tc subsets and potentiated the regulatory counterpart in T1D PBMCs, with a central role played by HLA-G5 in maintainining immunomodulation. . In conclusion, hUCMS in CpS may represent a functional biohybrid artificial delivery system, where molecular products induce efficient immunomodulation in T1D.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
