IFN-Gamma and TNF-Alpha as a Priming Strategy to Enhance the Immunomodulatory Capacity of Secretomes from Menstrual Blood-Derived Stromal Cells.

María Ángeles De Pedro,Christian Preußer,Verónica Álvarez,María Gómez-Serrano,María Pulido,Esther López,Federica Marinaro,Javier G Casado,Elke Pogge Von Strandmann,Francisco Miguel Sánchez-Margallo

doi:10.3390/ijms222212177

Abstract

Mesenchymal stromal cells isolated from menstrual blood (MenSCs) exhibit a potent pro-angiogenic and immunomodulatory capacity. Their therapeutic effect is mediated by paracrine mediators released by their secretomes. In this work, we aimed to evaluate the effect of a specific priming condition on the phenotype and secretome content of MenSCs. Our results revealed that the optimal condition for priming MenSCs was the combination of interferon gamma (IFNγ) and tumor necrosis factor alpha (TNFα) that produced a synergistic and additive effect on IDO1 release and immune-related molecule expression. The analyses of MenSC-derived secretomes after IFNγ and TNFα priming also revealed an increase in EV release and in the differentially expressed miRNAs involved in the immune response and inflammation. Proliferation assays on lymphocyte subsets demonstrated a decrease in CD4+ T cells and CD8+ T cells co-cultured with secretomes, especially in the lymphocytes co-cultured with secretomes from primed cells. Additionally, the expression of immune checkpoints (PD-1 and CTLA-4) was increased in the CD4+ T cells co-cultured with MenSC-derived secretomes. These findings demonstrate that the combination of IFNγ and TNFα represents an excellent priming strategy to enhance the immunomodulatory capacity of MenSCs. Moreover, the secretome derived from primed MenSCs may be postulated as a therapeutic option for the regulation of adverse inflammatory reactions.

Highlights

Human menstrual blood is an important source of stromal cells (MSCs)
We have evaluated different priming strategies to enhance the immunomodulatory capacity of secretomes from Menstrual blood-derived stromal cells (MenSCs)
Our MenSCs were plastic-adherent in standard culture conditions, and their differentiation toward the adipogenic, chondrogenic, and osteogenic lineages was demonstrated by specific staining

Summary

Introduction

Human menstrual blood is an important source of stromal cells (MSCs). Menstrual blood-derived stromal cells (MenSCs) fulfill the minimum criteria established by the International Society for Cellular Therapy: they exhibit plastic adherence in tissue and can be passaged in standard culture media; they express a CD105+, CD73+, CD90+, CD45−, CD34−, CD14−, and HLA-DR− phenotype; and they possess an in vitro differentiation capacity toward osteoblasts, adipocytes, and chondroblasts [1]. MenSCs are characterized by the well-known immunomodulatory and regenerative properties of MSCs, as well as immunosuppressive activity [2,3,4] and anti-apoptotic and pro-angiogenic capacities [5]. They stand out for their easy isolation, high proliferation rate, low immunogenicity, and lack of ethical issues compared with other sources of MSCs [6,7]. MSC-derived secretomes are composed of a set of MSC-derived bioactive factors in soluble form, along with those encapsulated in extracellular vesicles (EVs). They comprise proteins, nucleic acids, and lipids that are crucial for intercellular communication and responsible for MSCs’ therapeutic potential [1]

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