Generation of Retinal Pigment Epithelial Cells Derived from Human Embryonic Stem Cells Lacking Human Leukocyte Antigen Class I and II

Stem Cell Reports. 2020 Apr 14;14(4):648-662. doi: 10.1016/j.stemcr.2020.02.006. Epub 2020 Mar 19.


Human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells could serve as a replacement therapy in advanced stages of age-related macular degeneration. However, allogenic hESC-RPE transplants trigger immune rejection, supporting a strategy to evade their immune recognition. We established single-knockout beta-2 microglobulin (SKO-B2M), class II major histocompatibility complex transactivator (SKO-CIITA) and double-knockout (DKO) hESC lines that were further differentiated into corresponding hESC-RPE lines lacking either surface human leukocyte antigen class I (HLA-I) or HLA-II, or both. Activation of CD4+ and CD8+ T-cells was markedly lower by hESC-RPE DKO cells, while natural killer cell cytotoxic response was not increased. After transplantation of SKO-B2M, SKO-CIITA, or DKO hESC-RPEs in a preclinical rabbit model, donor cell rejection was reduced and delayed. In conclusion, we have developed cell lines that lack both HLA-I and -II antigens, which evoke reduced T-cell responses in vitro together with reduced rejection in a large-eyed animal model.

Keywords: HLA-I knockout; HLA-II knockout; cellular therapy; human embryonic stem cells; immune evasion; retinal pigment epithelium; subretinal injection; transplantation rejection; xenogeneic transplant; xenograft model.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Associated Protein 9 / metabolism
  • CRISPR-Cas Systems / genetics
  • Cytotoxicity, Immunologic
  • Epithelial Cells / metabolism*
  • Heterografts
  • Histocompatibility Antigens Class I / metabolism*
  • Histocompatibility Antigens Class II / metabolism*
  • Human Embryonic Stem Cells / cytology*
  • Human Embryonic Stem Cells / metabolism
  • Humans
  • Immunomodulation
  • Nuclear Proteins / metabolism
  • Polymorphism, Single Nucleotide / genetics
  • Retinal Pigment Epithelium / cytology*
  • T-Lymphocytes / metabolism
  • Trans-Activators / metabolism
  • beta 2-Microglobulin / metabolism


  • Histocompatibility Antigens Class I
  • Histocompatibility Antigens Class II
  • MHC class II transactivator protein
  • Nuclear Proteins
  • Trans-Activators
  • beta 2-Microglobulin
  • CRISPR-Associated Protein 9