Advertisement

Molecular Medicine

, Volume 21, Issue 1, pp 185–196 | Cite as

Human Parthenogenetic Embryonic Stem Cell-Derived Neural Stem Cells Express HLA-G and Show Unique Resistance to NK Cell-Mediated Killing

  • Jessica Schmitt
  • Sigrid Eckardt
  • Paul G. Schlegel
  • Anna-Leena Sirén
  • Valentin S. Bruttel
  • K. John McLaughlin
  • Jörg Wischhusen
  • Albrecht M. Müller
Research Article

Abstract

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.

Notes

Acknowledgments

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).

Supplementary material

10020_2015_2101185_MOESM1_ESM.pdf (1.9 mb)
Supplementary material, approximately 1.92 MB.

References

  1. 1.
    Revazova ES, et al. (2007) Patient-specific stem cell lines derived from human parthenogenetic blastocysts. Cloning Stem Cells. 9:432–49.CrossRefGoogle Scholar
  2. 2.
    Mai Q, et al. (2007) Derivation of human embryonic stem cell lines from parthenogenetic blastocysts. Cell Res. 17:1008–19.CrossRefGoogle Scholar
  3. 3.
    Lin G, et al. (2007) A highly homozygous and parthenogenetic human embryonic stem cell line derived from a one-pronuclear oocyte following in vitro fertilization procedure. Cell Res. 17:999–1007.CrossRefGoogle Scholar
  4. 4.
    Kim K, et al. (2007) Recombination signatures distinguish embryonic stem cells derived by parthenogenesis and somatic cell nuclear transfer. Cell Stem Cell. 1:346–52.CrossRefGoogle Scholar
  5. 5.
    Brevini TA, et al. (2009) Cell lines derived from human parthenogenetic embryos can display aberrant centriole distribution and altered expression levels of mitotic spindle check-point transcripts. Stem Cell Rev. 5:340–52.CrossRefGoogle Scholar
  6. 6.
    Revazova ES, et al. (2008) HLA homozygous stem cell lines derived from human parthenogenetic blastocysts. Cloning Stem Cells. 10:11–24.CrossRefGoogle Scholar
  7. 7.
    Turovets N, et al. (2011) Derivation of human parthenogenetic stem cell lines. Methods Mol. Biol. 767:37–54.CrossRefGoogle Scholar
  8. 8.
    Daughtry B, Mitalipov S. (2014) Concise review: parthenote stem cells for regenerative medicine: genetic, epigenetic, and developmental features. Stem Cells Transl. Med. 3:290–8.CrossRefGoogle Scholar
  9. 9.
    Wilkinson LS, Davies W, Isles AR. (2007) Genomic imprinting effects on brain development and function. Nat. Rev. Neurosci. 8:832–43.CrossRefGoogle Scholar
  10. 10.
    Keverne EB, Fundele R, Narasimha M, Barton SC, Surani MA. (1996) Genomic imprinting and the differential roles of parental genomes in brain development. Brain Res. Dev. Brain Res. 92:91–100.CrossRefGoogle Scholar
  11. 11.
    Dinger TC, et al. (2008) Androgenetic embryonic stem cells form neural progenitor cells in vivo and in vitro. Stem Cells. 26:1474–83.CrossRefGoogle Scholar
  12. 12.
    Ahmad R, et al. (2012) Functional neuronal cells generated by human parthenogenetic stem cells. PLoS One. 7:e42800.CrossRefGoogle Scholar
  13. 13.
    Choi SW, et al. (2010) Two paternal genomes are compatible with dopaminergic in vitro and in vivo differentiation. Int. J. Dev. Biol. 54:1755–62.CrossRefGoogle Scholar
  14. 14.
    Wolber W, et al. (2013) Phenotype and stability of neural differentiation of androgenetic murine ES cell-derived neural progenitor cells. Cell Medicine. 5:29–42.CrossRefGoogle Scholar
  15. 15.
    Pearl JI, Kean LS, Davis MM, Wu JC. (2012) Pluripotent stem cells: immune to the immune system? Sci. Transi. Med. 4:164ps125.Google Scholar
  16. 16.
    Drukker M, et al. (2002) Characterization of the expression of MHC proteins in human embryonic stem cells. Proc. Natl. Acad. Sci. USA. 99:9864–9.CrossRefGoogle Scholar
  17. 17.
    Grinnemo KH, et al. (2006) Human embryonic stem cells are immunogenic in allogeneic and xenogeneic settings. Reprod. Biomed. Online. 13:712–24.CrossRefGoogle Scholar
  18. 18.
    Li L, et al. (2004) Human embryonic stem cells possess immune-privileged properties. Stem Cells. 22:448–56.CrossRefGoogle Scholar
  19. 19.
    Preynat-Seauve O, et al. (2009) Neural progenitors derived from human embryonic stem cells are targeted by allogeneic T and natural killer cells. J. Cell. Mol. Med. 13:3556–69.CrossRefGoogle Scholar
  20. 20.
    Liu J, et al. (2013) Human neural stem/progenitor cells derived from embryonic stem cells and fetal nervous system present differences in immunogenicity and immunomodulatory potentials in vitro. Stem Cell Res. 10:325–37.CrossRefGoogle Scholar
  21. 21.
    Drukker M, et al. (2006) Human embryonic stem cells and their differentiated derivatives are less susceptible to immune rejection than adult cells. Stem Cells. 24:221–9.CrossRefGoogle Scholar
  22. 22.
    Tian X, Woll PS, Morris JK, Linehan JL, Kaufman DS. (2006) Hematopoietic engraftment of human embryonic stem cell-derived cells is regulated by recipient innate immunity. Stem Cells. 24:1370–80.CrossRefGoogle Scholar
  23. 23.
    Rouas-Freiss N, Goncalves RM, Menier C, Dausset J, Carosella ED. (1997) Direct evidence to support the role of HLA-G in protecting the fetus from maternal uterine natural killer cytolysis. Proc. Natl. Acad. Sci. USA. 94:11520–5.CrossRefGoogle Scholar
  24. 24.
    Carosella ED, Favier B, Rouas-Freiss N, Moreau P, Lemaoult J. (2008) Beyond the increasing complexity of the immunomodulatory HLA-G molecule. Blood. 111:4862–70.CrossRefGoogle Scholar
  25. 25.
    Jurisicova A, Casper RF, MacLusky NJ, Mills GB, Librach CL. (1996) HLA-G expression during preimplantation human embryo development. Proc. Natl. Acad. Sci. USA. 93:161–5.CrossRefGoogle Scholar
  26. 26.
    Maier S, Geraghty DE, Weiss EH. (1999) Expression and regulation of HLA-G in human glioma cell lines. Transplant. Proc. 31:1849–53.CrossRefGoogle Scholar
  27. 27.
    Lafon M, et al. (2005) Modulation of HLA-G expression in human neural cells after neurotropic viral infections. J. Virol. 79:15226–37.CrossRefGoogle Scholar
  28. 28.
    Nasef A, et al. (2007) Immunosuppressive effects of mesenchymal stem cells: involvement of HLA-G. Transplantation. 84:231–7.CrossRefGoogle Scholar
  29. 29.
    Wiendl H. (2007) HLA-G in the nervous system. Hum. Immunol. 68:286–93.CrossRefGoogle Scholar
  30. 30.
    LeMaoult J, et al. (2003) Biology and functions of human leukocyte antigen-G in health and sickness. Tissue Antigens. 62:273–84.CrossRefGoogle Scholar
  31. 31.
    Verloes A, et al. (2011) HLA-G expression in human embryonic stem cells and preimplantation embryos. J. Immunol. 186:2663–71.CrossRefGoogle Scholar
  32. 32.
    Le Page ME, Goodridge JP, John E, Christiansen FT, Witt CS. (2014) Killer Ig-like receptor 2DL4 does not mediate NK cell IFN-gamma responses to soluble HLA-G preparations. J. Immunol. 192:732–40.CrossRefGoogle Scholar
  33. 33.
    Thomson JA, et al. (1998) Embryonic stem cell lines derived from human blastocysts. Science. 282:1145–7.CrossRefGoogle Scholar
  34. 34.
    Ström S, Holm F, Bergstrom R, Stromberg AM, Hovatta O. (2010) Derivation of 30 human embryonic stem cell lines-improving the quality. In Vitro Cell. Dev. Biol. Anim. 46:337–44.CrossRefGoogle Scholar
  35. 35.
    Fürst D, et al. (2013) High-resolution HLA matching in hematopoietic stem cell transplantation: a retrospective collaborative analysis. Blood. 122:3220–9.CrossRefGoogle Scholar
  36. 36.
    Zhu XM, et al. (2010) Overexpression of miR-152 leads to reduced expression of human leukocyte antigen-G and increased natural killer cell mediated cytolysis in JEG-3 cells. Am. J. Obstet. Gynecol. 202: 592.e1–7.Google Scholar
  37. 37.
    Ji W, et al. (2013) MicroRNA-152 targets DNA methyltransferase 1 in NiS-transformed cells via a feedback mechanism. Carcinogenesis. 34:446–53.CrossRefGoogle Scholar
  38. 38.
    Rohde C, Zhang Y, Reinhardt R, Jeltsch A. (2010) BISMA-fast and accurate bisulfite sequencing data analysis of individual clones from unique and repetitive sequences. BMC Bioinformatics. 11:230.CrossRefGoogle Scholar
  39. 39.
    Burt D, Johnston D, Rinke de Wit T, Van den Elsen P, Stern PL. (1991) Cellular immune recognition of HLA-G-expressing choriocarcinoma cell line Jeg-3. Int. J. Cancer Suppl. 6:117–22.CrossRefGoogle Scholar
  40. 40.
    Ljunggren HGK, K. (1990) In search of the “missing self”: MHC molecules and NK cell recognition. Immunol. Today. 11:237–44.CrossRefGoogle Scholar
  41. 41.
    Zhu Y, et al. (2012) DA neurons derived from hES cells that express HLA-G1 are capable of immunosuppression. Brain Res. 1437:134–42.CrossRefGoogle Scholar
  42. 42.
    Manaster I, et al. (2012) MiRNA-mediated control of HLA-G expression and function. PLoS One. 7:e33395.CrossRefGoogle Scholar
  43. 43.
    Bock C, et al. (2011) Reference maps of human ES and iPS cell variation enable high-throughput characterization of pluripotent cell lines. Cell. 144:439–52.CrossRefGoogle Scholar
  44. 44.
    Hiby SE, King A, Sharkey A, Loke YW. (1999) Molecular studies of trophoblast HLA-G: polymorphism, isoforms, imprinting and expression in preimplantation embryo. Tissue Antigens. 53:1–13.CrossRefGoogle Scholar
  45. 45.
    Hviid TVF, Moller C, Sorensen S, Morling N. (1998) Co-dominant expression of the HLA-G gene and various forms of alternatively spliced HLA-G mRNA in human first trimester trophoblast. Hum. Immunology. 59:87–98.CrossRefGoogle Scholar
  46. 46.
    Zheng X, Chopp M, Lu Y, Buller B, Jiang F. (2013) MiR-15b and miR-152 reduce glioma cell invasion and angiogenesis via NRP-2 and MMP-3. Cancer Lett. 329:146–54.CrossRefGoogle Scholar
  47. 47.
    Veit TD, Chies JA. (2009) Tolerance versus immune response — microRNAs as important elements in the regulation of the HLA-G gene expression. Transpl. Immunol. 20:229–31.CrossRefGoogle Scholar
  48. 48.
    Menier C, et al. (2004) Erythroblasts secrete the nonclassical HLA-G molecule from primitive to definitive hematopoiesis. Blood 104:3153–60.CrossRefGoogle Scholar
  49. 49.
    Stridh P, et al. (2014) Parent-of-origin effects implicate epigenetic regulation of experimental autoimmune encephalomyelitis and identify imprinted Dlk1 as a novel risk gene. PLoS. Genet. 10:e1004265.CrossRefGoogle Scholar
  50. 50.
    Elmer BM, McAllister AK. (2012) Major histocompatibility complex class I proteins in brain development and plasticity. Trends Neurosci. 35:660–70.CrossRefGoogle Scholar
  51. 51.
    Wiendl H, et al. (2005) Expression of the immune-tolerogenic major histocompatibility molecule HLA-G in multiple sclerosis: implications for CNS immunity. Brain. 128:2689–704.CrossRefGoogle Scholar
  52. 52.
    Hickey WF. (2001) Basic principles of immunological surveillance of the normal central nervous system. Glia. 36:118–24.CrossRefGoogle Scholar
  53. 53.
    Yen BL, et al. (2009) Brief report—human embryonic stem cell-derived mesenchymal progenitors possess strong immunosuppressive effects toward natural killer cells as well as T lymphocytes. Stem Cells. 27:451–6.CrossRefGoogle Scholar
  54. 54.
    Pluchino S, Cossetti C. (2013) How stem cells speak with host immune cells in inflammatory brain diseases. Glia. 61:1379–401.CrossRefGoogle Scholar
  55. 55.
    Bommireddy R, et al. (2009) Calcineurin deficiency decreases inflammatory lesions in transforming growth factor beta1-deficient mice. Clin. Exp. Immunol. 158:317–24.CrossRefGoogle Scholar
  56. 56.
    Ubiali F, et al. (2007) Allorecognition of human neural stem cells by peripheral blood lymphocytes despite low expression of MHC molecules: role of TGF-beta in modulating proliferation. Int. Immunol. 19:1063–74.CrossRefGoogle Scholar
  57. 57.
    Bonnamain V, et al. (2012) Expression of heme oxygenase-1 in neural stem/progenitor cells as a potential mechanism to evade host immune response. Stem Cells. 30:2342–53.CrossRefGoogle Scholar
  58. 58.
    Wang L, et al. (2009) Neural stem/progenitor cells modulate immune responses by suppressing T lymphocytes with nitric oxide and prostaglandin E2. Exp. Neurol. 216:177–83.CrossRefGoogle Scholar
  59. 59.
    Carletti B, Piemonte F, Rossi F. (2011) Neuroprotection: the emerging concept of restorative neural stem cell biology for the treatment of neurodegenerative diseases. Curr. Neuropharmacol. 9:313–7.CrossRefGoogle Scholar
  60. 60.
    Lindvall O, Barker RA, Brustle O, Isacson O, Svendsen CN. (2012) Clinical translation of stem cells in neurodegenerative disorders. Cell Stem Cell. 10:151–5.CrossRefGoogle Scholar

Copyright information

© The Author(s) 2015

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (https://doi.org/creativecommons.org/licenses/by-nc-nd/4.0/)

Authors and Affiliations

  • Jessica Schmitt
    • 1
  • Sigrid Eckardt
    • 2
  • Paul G. Schlegel
    • 3
  • Anna-Leena Sirén
    • 4
  • Valentin S. Bruttel
    • 5
  • K. John McLaughlin
    • 2
  • Jörg Wischhusen
    • 5
  • Albrecht M. Müller
    • 1
  1. 1.Institute for Medical Radiology and Cell Research (MSZ) in the Center for Experimental Molecular Medicine (ZEMM)University of WürzburgWürzburgGermany
  2. 2.Center for Molecular and Human GeneticsThe Research Institute at Nationwide Children’s HospitalColumbusUSA
  3. 3.Pediatric Hematology/OncologyUniversity Children’s Hospital WürzburgWürzburgGermany
  4. 4.Department of NeurosurgeryUniversity of WürzburgWürzburgGermany
  5. 5.University of Würzburg Medical School, Department of Obstetrics and Gynecology, Section for Experimental Tumor ImmunologyUniversity of WürzburgWürzburgGermany

Personalised recommendations