Antigen Presentation by Individually Transferred HLA Class I Genes in HLA-A, HLA-B, HLA-C Null Human Cell Line Generated Using the Multiplex CRISPR-Cas9 System

• Published in: Journal of immunotherapy (1997) Vol. 40; no. 6; p. 201
• Main Authors: Hong, Cheol-Hwa, Sohn, Hyun-Jung, Lee, Hyun-Joo, Cho, Hyun-Il, Kim, Tai-Gyu
• Format: Journal Article
• Language: English
• Published: United States 01.07.2017
• Subjects: Antigen-Presenting Cells - physiology; Antigen-Presenting Cells - transplantation; Antigens, Neoplasm - immunology; Antigens, Neoplasm - metabolism; Cell Line; CRISPR-Cas Systems; Exons - genetics; Gene Editing; Graft Rejection - immunology; Graft Rejection - prevention & control; Graft vs Host Disease - immunology; Graft vs Host Disease - prevention & control; Hematopoietic Stem Cell Transplantation; HLA-A Antigens - genetics; HLA-A Antigens - metabolism; HLA-B Antigens - genetics; HLA-B Antigens - metabolism; HLA-C Antigens - genetics; HLA-C Antigens - metabolism; Humans; Immunotherapy, Adoptive - methods; Lymphocyte Activation; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - transplantation
• ISSN: 1537-4513
• DOI: 10.1097/cji.0000000000000176

Human leukocyte antigens (HLAs) are essential immune molecules that affect transplantation and adoptive immunotherapy. When hematopoietic stem cells or organs are transplanted with HLA-mismatched recipients, graft-versus-host disease or graft rejection can be induced by allogeneic immune responses. The function of each HLA allele has been studied using HLA-deficient cells generated from mutant cell lines or by RNA interference, zinc finger nuclease, and the CRISPR/Cas9 system. To improve HLA gene editing, we attempted to generate an HLA class I null cell line using the multiplex CRISPR/Cas9 system by targeting exons 2 and 3 of HLA-A, HLA-B, and HLA-C genes simultaneously. Multiplex HLA editing could induce the complete elimination of HLA class I genes by bi-allelic gene disruption on target sites which was defined by flow cytometry and target-specific polymerase chain reaction. Furthermore, artificial antigen-presenting cells were generated by transfer of a single HLA class I allele and co-stimulatory molecules into this novel HLA class I null cell line. Artificial antigen-presenting cells showed HLA-restricted antigen presentation following antigen processing and were successfully used for the efficient generation of tumor antigen-specific cytotoxic T cells in vitro. The efficient editing of HLA genes may provide a basis for universal cellular therapies and transplantation.