Functional disruption of human leukocyte antigen II in human embryonic stem cell.
BACKGROUND
Theoretically human embryonic stem cells (hESCs) have the capacity to self-renew and differentiate into all human cell types. Therefore, the greatest promise of hESCs-based therapy is to replace the damaged tissues of patients suffering from traumatic or degenerative diseases by the exact same type of cells derived from hESCs. Allograft immune rejection is one of the obstacles for hESCs-based clinical applications. Human leukocyte antigen (HLA) II leads to CD4(+) T cells-mediated allograft rejection. Hence, we focus on optimizing hESCs for clinic application through gene modification.


RESULTS
Transcription activator-like effector nucleases (TALENs) were used to target MHC class II transactivator (CIITA) in hESCs efficiently. CIITA (-/-) hESCs did not show any difference in the differentiation potential and self-renewal capacity. Dendritic cells (DCs) derived from CIITA (-/-) hESCs expressed CD83 and CD86 but without the constitutive HLA II. Fibroblasts derived from CIITA (-/-) hESCs were powerless in IFN-γ inducible expression of HLA II.


CONCLUSIONS
We generated HLA II defected hESCs via deleting CIITA, a master regulator of constitutive and IFN-γ inducible expression of HLA II genes. CIITA (-/-) hESCs can differentiate into tissue cells with non-HLA II expression. It's promising that CIITA (-/-) hESCs-derived cells could be used in cell therapy (e.g., T cells and DCs) and escape the attack of receptors' CD4(+) T cells, which are the main effector cells of cellular immunity in allograft.
College of Animal Science, Zhejiang University, Hangzhou, 310058, People's Republic of China. Hyde@zju.edu.cn.|Zhejiang University School of Medicine, Hangzhou, 310058, People's Republic of China. Hyde@zju.edu.cn.
Haide
H
Chen
College of Animal Science, Zhejiang University, Hangzhou, 310058, People's Republic of China. Younglee@zju.edu.cn.|Zhejiang University School of Medicine, Hangzhou, 310058, People's Republic of China. Younglee@zju.edu.cn.
Yang
Y
Li
College of Animal Science, Zhejiang University, Hangzhou, 310058, People's Republic of China. 11217009@zju.edu.cn.|Zhejiang University School of Medicine, Hangzhou, 310058, People's Republic of China. 11217009@zju.edu.cn.
Xijuan
X
Lin
College of Animal Science, Zhejiang University, Hangzhou, 310058, People's Republic of China. 21217007@zju.edu.cn.|Zhejiang University School of Medicine, Hangzhou, 310058, People's Republic of China. 21217007@zju.edu.cn.
Di
D
Cui
Wuxi Medical School, Jiangnan University, Wuxi, 214122, People's Republic of China. Cuichun@jiangnan.edu.cn.
Chun
C
Cui
Xiangtan Center Hospital, Hunan, 411100, People's Republic of China. Lihui_1973@126.com.
Hui
H
Li
College of Animal Science, Zhejiang University, Hangzhou, 310058, People's Republic of China. Leixiao@zju.edu.cn.|Zhejiang University School of Medicine, Hangzhou, 310058, People's Republic of China. Leixiao@zju.edu.cn.
Lei
L
Xiao
D000818:Animals; D000938:Antigen-Presenting Cells / Q000378:metabolism; D015703:Antigens, CD / Q000378:metabolism; D051940:B7-2 Antigen / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D000066673:Cell Self Renewal; D003713:Dendritic Cells / Q000378:metabolism; D003851:Deoxyribonucleases / Q000145:classification / Q000378:metabolism*; D058732:Embryoid Bodies / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D017353:Gene Deletion; D000949:Histocompatibility Antigens Class II / Q000235:genetics; D000066449:Human Embryonic Stem Cells / Q000378:metabolism*; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism; D007150:Immunohistochemistry; D007371:Interferon-gamma / Q000378:metabolism; D059785:Karyotype; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D009687:Nuclear Proteins / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013724:Teratoma; D015534:Trans-Activators / Q000235:genetics*; D014407:Tumor Cells, Cultured; D000097802:CD83 Antigen
D015703:Antigens, CD; D051940:B7-2 Antigen; D003851:Deoxyribonucleases; D000949:Histocompatibility Antigens Class II; D007136:Immunoglobulins; D007371:Interferon-gamma; D008562:Membrane Glycoproteins; D009687:Nuclear Proteins; D015534:Trans-Activators; D000097802:CD83 Antigen; C083728:MHC class II transactivator protein