Human Parthenogenetic Embryonic Stem Cell-Derived Neural Stem Cells Express HLA-G and Show Unique Resistance to NK Cell-Mediated Killing
Parent-of-origin imprints have been implicated in the regulation of neural differentiation and brain development. Previously we have shown that, despite the lack of a paternal genome, human parthenogenetic (PG) embryonic stem cells (hESCs) can form proliferating neural stem cells (NSCs) that are capable of differentiation into physiologically functional neurons while maintaining allele-specific expression of imprinted genes. Since biparental (“normal”) hESC-derived NSCs (N NSCs) are targeted by immune cells, we characterized the immunogenicity of PG NSCs. Flow cytometry and immunocytochemistry revealed that both N NSCs and PG NSCs exhibited surface expression of human leukocyte antigen (HLA) class I but not HLA-DR molecules. Functional analyses using an in vitro mixed lymphocyte reaction assay resulted in less proliferation of peripheral blood mononuclear cells (PBMC) with PG compared with N NSCs. In addition, natural killer (NK) cells cytolyzed PG less than N NSCs. At a molecular level, expression analyses of immune regulatory factors revealed higher HLA-G levels in PG compared with N NSCs. In line with this finding, MIR152, which represses HLA-G expression, is less transcribed in PG compared with N cells. Blockage of HLA-G receptors ILT2 and KIR2DL4 on natural killer cell leukemia (NKL) cells increased cytolysis of PG NSCs. Together this indicates that PG NSCs have unique immunological properties due to elevated HLA-G expression.
We thank Ruslan Semechkin for kindly providing the PG hESCs LLC6P and LLC9P (International Stem Cell Corporation), Outi Hovatta and Liselotte Antonsson (Division of Obstetrics and Gynecology, Karolinska Institutet, Stockholm, Sweden) for the HS401 hESCs, WiCell Research Institute Wisconsin (Madison, USA) for the H9 hESCs, Ulrike Kämmerer (Department of Obstetrics and Gynecology, University of Würzburg, Germany) for the JEG-3 cells and Winfried S Wels (Georg-Speyer-Haus, Frankfurt am Main, Germany) for the NKL cell line. We are grateful to Andrea Reusch and Doris Heim (MSZ, Würzburg, Germany) for technical assistance, and to Andrea Niklaus and Katharina Mattenheimer (ZEMM, Würzburg, Germany) for help with sample collection. We thank Joannis Mytilineos (Institute of Clinical Transfusion Medicine and Immunogenetics Ulm, Germany) for high-resolution HLA typing. Funding for this work was provided by the Interdisciplinary Center for Clinical Research (IZKF), University of Würzburg (TP D103), DFG-funded SPP 1738 and by a fellowship, Chancengleichheit für Frauen in Forschung und Lehre, from the University of Würzburg (Würzburg, Germany).
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