Identification of Potential Immunogenic Molecules During the Allogeneic Transplantation of Human Adipose-derived Mesenchymal Stem Cells
Abstract Background: Given their low immunogenicity and multiple differentiation capacities, mesenchymal stem cells (MSCs) have the potential to be used for “off-the-shelf” cell therapy. However, MSC allorejection indicates that they are not fully immune privileged. In this study, we investigated the immunogenicity of human adipose-derived MSCs (Ad-MSCs) and identified potential immunogenic molecules.Methods: To evaluate the immunogenicity of human Ad-MSCs in vivo, cells were transplanted into humanized mice (hu-mice), and T cell infiltration and clearance of human Ad-MSCs were observed by immunofluorescence and bioluminescence imaging. One-way mixed lymphocyte reaction (MLR) and flow cytometry were performed to evaluate the immunogenicity of human Ad-MSCs in vitro. High-throughput TCR repertoire sequencing and mass spectrometry were applied to identify potential immunogenic molecules.Results: Allogeneic human Ad-MSCs recruited T cells during transplantation and caused faster clearance in hu-mice than NOD/ShiLtJGpt-Prkdcem26Cd52Il2rgem26Cd22/Gpt (NSG) mice. The proliferation and activation of T cells was significantly enhanced by human Ad-MSCs, and the expression level of HLA-II on human Ad-MSCs was dramatically increased after coculture with human peripheral blood mononuclear cells (PBMCs) in vitro. In addition, upregulated expression of alpha-enolase (ENO1) on the surface of human Ad-MSCs increased their immunogenicity, and ENO1 inhibitor treatment decreased the human Ad-MSC triggered proliferation of T cells in vitro.Conclusions: We further confirmed the immunogenicity of human Ad-MSCs during allogeneic transplantation and provided a potential target, ENO1, for the safe clinical application of allogeneic human Ad-MSC therapy.