Amniotic membrane extract differentially regulates human peripheral blood T cell subsets, monocyte subpopulations and myeloid dendritic cells
The discovery of the immunoregulatory potential of human amniotic membrane (hAM) propelled several studies focusing on its application for the treatment of immunological disorders. However, there is little information regarding the effects of hAM on distinct activation and differentiation stages of immune cells. Here, we aim to investigate the effect of human amniotic membrane extract (hAME) on the pattern of cytokine production by T cells, monocytes and myeloid dendritic cells (mDCs). For this purpose, peripheral blood mononuclear cells (PBMCs) from eight healthy individuals were stimulated in vitro in the presence or absence of hAME. Mitogen-induced proliferation of PBMCs and cytokine production among the distinct T cell functional compartments, monocyte subpopulations and mDCs were evaluated. hAME displayed an anti-proliferative effect and decreased the frequency of T cells producing tumor necrosis factor (TNF)α, interferon (IFN)γ and interleukin (IL)-2, for all T cell functional compartments. The frequency of IL-17 and IL-9-producing T cells was also reduced. The inhibition of mRNA expression of granzyme B, perforin and NKG2D by CD8+ T cells and γδ T cells and the augment of FoxP3 and IL-10 in CD4+ T cells and IL-10 in regulatory T cells were also observed. Furthermore, hAME inhibited IFNγ-induced protein (IP)-10 expression by classical and non-classical monocytes, without hampering the production of TNFα and IL-6 by monocytes and mDCs. These results suggest that hAME exerts an anti-inflammatory effect on T cells, still at a different extent for distinct T cell functional compartments.
KeywordsHuman amniotic membrane Immunosuppression T cell Monocyte Dendritic cell
Ana Catarina Mamede (SFRH/BD/73649/2010) and Maria João Carvalho (SFRH/SINTD/60068/2009) wish to thank the Fundação para a Ciência e a Tecnologia (FCT, Portugal) for the PhD grants. The authors would also like to thank the Obstetrics Service of the Centro Hospitalar e Universitário de Coimbra for the collection of human tissues used in this work.
Ana Catarina Mamede and Maria João Carvalho received PhD grants (SFRH/BD/73649/2010 and SFRH/SINTD/60068/2009, respectively) by Fundação para a Ciência e a Tecnologia (FCT), Portugal. This study was funded by Infarmed (Health Research Fund 2015, FIS-2015-01), Portugal.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The study was approved by the Ethical Committee of the Centro Hospitalar e Universitário de Coimbra (CHUC-70-12, Coimbra, Portugal). All procedures performed in the studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments.
Informed consent was obtained from all individual participants included in the study.
- Guggino G, Lo Pizzo M, Di Liberto D, Rizzo A, Cipriani P, Ruscitti P, Candore G, Gambino CM, Sireci G, Dieli F, Giacomelli R, Triolo G, Ciccia F (2017) Interleukin-9 over-expression and t helper 9 polarization in systemic sclerosis patients. Clin Exp ImmunolGoogle Scholar
- He H, Li W, Tseng DY, Zhang S, Chen SY, Day AJ, Tseng SC (2009) Biochemical characterization and function of complexes formed by hyaluronan and the heavy chains of inter-alpha-inhibitor (hc*ha) purified from extracts of human amniotic membrane. J Biol Chem 284:20136–20146CrossRefPubMedPubMedCentralGoogle Scholar
- Li J, Chen S, Xiao X, Zhao Y, Ding W, Li XC (2017) IL-9 and Th9 cells in health and diseases—from tolerance to immunopathology. Cytokine Growth Factor RevGoogle Scholar
- Magatti M, Vertua E, De Munari S, Caro M, Caruso M, Silini A, Delgado M, Parolini O (2016) Human amnion favours tissue repair by inducing the m1-to-m2 switch and enhancing m2 macrophage features. J Tissue Eng Regen MedGoogle Scholar
- Ziegler-Heitbrock L, Ancuta P, Crowe S, Dalod M, Grau V, Hart DN, Leenen PJ, Liu YJ, MacPherson G, Randolph GJ, Scherberich J, Schmitz J, Shortman K, Sozzani S, Strobl H, Zembala M, Austyn JM, Lutz MB (2010) Nomenclature of monocytes and dendritic cells in blood. Blood 116:e74–e80CrossRefPubMedGoogle Scholar