Abstract
Human amniotic cells (hAC) possess multiple unique immunomodulatory properties. They are believed to be a very appealing and safe material for clinical applications. Primary hAC have been proposed as an efficient source of immunomodulatory factors that could be used as alternative or supporting to classical drug immunosuppression. The aim of this study was to evaluate hAC immunomodulatory properties post-activation by inflammatory cytokines as Interleukin 1β and Interferon γ. hAC were isolated and characterized by the expression of pluripotency marker SSEA4, epithelial marker CK7, HLA-G antigen, mRNA for PTGS2, NOS2 and HLA-G gene, and secretion of soluble mediators as HLA-G and PGE2 in the culture medium in presence or absence of INF-γ and IL-1β. Heterogeneity of the cultured hAC was proved, with 50 ± 8% of cells positive for epithelial marker (CK7), and 73 ± 3% expressing SSEA4 pluripotency marker. Priming effect by in vitro exposure to INF-γ and IL-1β resulted in a significant increase in expression of PTGS2, NOS2 and HLA-G gene, with a peak between 32 and 64 h. The highest PGE2 concentration was measured in the culture medium at 48 h. At 96 h, a significant difference in the percentage of SSEA4+ hAC between activated and non-activated cells, as well as the highest expression of HLA-G - especially in SSEA4+ cells, and highly elevated concentration of soluble HLA-G (sHLA-G) in the medium of activated cells, were found. The prolonged exposure of primary human amnion-derived cells to inflammatory cytokines INF-γ and IL-1β may result in enhanced expression and secretion of immunomodulatory molecules important in allogenic therapies.