Magatti Marta, Caruso Maddalena, De Munari Silvia, Vertua Elsa, De Debashree, Manuelpillai Ursula, Parolini Ornella
Centro di Ricerca E. Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy.
Cell Transplant. 2015;24(9):1733-52. doi: 10.3727/096368914X684033. Epub 2014 Aug 19.
We previously demonstrated that mesenchymal cells from human amniotic membrane (hAMTCs) inhibit the generation and maturation of monocyte-derived dendritic cells (DCs) in vitro. Considering the crucial role of DCs in the immune response and that epithelial cells of the human amniotic membrane (hAECs) share some of the immunoregulatory properties of hAMTCs, we investigated whether hAECs also modulate monocyte-derived DCs. We compared hAECs with hAMTCs in a cell-to-cell contact setting and their secreted factors in modulating DC differentiation and function. First, we demonstrated that primary and expanded hAMTCs strongly inhibited the differentiation of DCs and induced a shift toward M2-like macrophages. This was observed when hAMTCs were cultured in contact (hAMTC-DC(cont)) or in Transwells (hAMTC-DC(tw)) with monocytes and even when medium conditioned by hAMTCs was used instead of hAMTCs. hAECs also prevented DC development, but to a lesser extent than hAMTCs. hAECs were more effective when cultured in contact with monocytes (hAEC-DC(cont)) rather than in Transwells (hAEC-DC(tw)). The modulatory capacity of hAECs changed during passaging unlike the hAMSCs. The ability to stimulate CD4(+) and CD8(+) T-cell proliferation was almost completely abolished by hAMTC-DC(cont), whereas hAMTC-DC(tw) and hAEC-DC(cont) displayed only a reduced ability to stimulate CD8(+) T cells. Furthermore, monocytes cocultured with hAMTCs and hAECs showed some similarities, but also differences in cytokine/chemokine secretion. Similarities were observed in the inhibition of IL-12p70 and TNF-α and the increase in IL-10 in supernatants taken from monocyte-DCs cocultured with hAMTCs and hAECs in contact and Transwell settings. The inflammatory factors IL-8, CXCL9, and MIP-1α were significantly lower in hAMTC-DC(cont), hAMTC-DC(tw), and hAEC-DC(cont) conditions. In contrast, only hAMTCs (in both contact and Transwell conditions) were able to significantly increase IL-1β and CCL2. Altogether, we demonstrated that hAMTCs and hAECs affect DC differentiation, but that hAMTCs exerted a stronger inhibitory effect, abolished T-cell proliferation, and also induced more changes in cytokine/chemokine production.
我们之前证明，人羊膜间充质细胞（hAMTCs）在体外可抑制单核细胞来源的树突状细胞（DCs）的生成和成熟。鉴于DCs在免疫反应中的关键作用，以及人羊膜上皮细胞（hAECs）具有hAMTCs的一些免疫调节特性，我们研究了hAECs是否也能调节单核细胞来源的DCs。我们在细胞间接触环境中比较了hAECs和hAMTCs，以及它们在调节DC分化和功能方面的分泌因子。首先，我们证明原代和扩增的hAMTCs强烈抑制DCs的分化，并诱导其向M2样巨噬细胞转变。当hAMTCs与单核细胞在接触培养（hAMTC-DC(cont)）或Transwell培养（hAMTC-DC(tw)）中培养时，甚至当使用hAMTCs条件培养基代替hAMTCs时，都观察到了这种现象。hAECs也能阻止DC的发育，但程度比hAMTCs小。hAECs与单核细胞接触培养（hAEC-DC(cont)）时比在Transwell培养（hAEC-DC(tw)）时更有效。与hAMSCs不同，hAECs的调节能力在传代过程中发生了变化。hAMTC-DC(cont)几乎完全消除了刺激CD4(+)和CD8(+) T细胞增殖的能力，而hAMTC-DC(tw)和hAEC-DC(cont)仅表现出刺激CD8(+) T细胞的能力降低。此外，与hAMTCs和hAECs共培养的单核细胞在细胞因子/趋化因子分泌方面表现出一些相似性，但也存在差异。在接触培养和Transwell培养环境下，与hAMTCs和hAECs共培养的单核细胞-DCs培养上清中，IL-12p70和TNF-α的分泌受到抑制，IL-10的分泌增加，观察到了相似性。在hAMTC-DC(cont)、hAMTC-DC(tw)和hAEC-DC(cont)条件下，炎症因子IL-8、CXCL9和MIP-1α显著降低。相反，只有hAMTCs（在接触培养和Transwell培养条件下）能够显著增加IL-1β和CCL2。总之，我们证明hAMTCs和hAECs会影响DC分化，但hAMTCs具有更强的抑制作用，消除了T细胞增殖，并且在细胞因子/趋化因子产生方面也诱导了更多变化。
