Artificial antigen-presenting cell prepared from HLA-null cell line by using multiplex CRISPR-CAS9 system and use thereof
The present invention relates to an artificial antigen-presenting cell prepared from an HLA-null cell line by using a multiplex CRISPR-Cas9 system and the use thereof and, more particularly, to a novel artificial antigen-presenting cell which includes the ability to present antigens of HLA class I and a co-stimulatory molecule group transferred from an HLA-A, -B, -C null cell line generated using a multiplex CRISPR-Cas9 system and to stimulate T cells, an immunotherapeutic agent using the same, and the use thereof for treating tumors, pathogenic infections, and autoimmune diseases.
1. A method to make one or more antigen-presenting cells, comprising:
a) eliminating HLA-A, HLA-B and HLA-C in one or more 293T cells using a CRISPR-Cas9 system to make one or more HLA-null 293T cells,
wherein the CRISPR-Cas9 system comprises single guide RNAs(sgRNAs) targeting each of:
HLA-A exon 2(SEQ ID NO: 1) and exon 3(SEQ ID NO: 2),
HLA-B exon 2(SEQ ID NO: 3) and exon 3(SEQ ID NO: 4),
HLA-C exon 2(SEQ ID NO: 5) and exon 3(SEQ ID NO: 6); and
b) introducing a co-stimulatory molecule group into the one or more HLA-null 293T cells to make one or more antigen-presenting cells,
wherein the co-stimulatory molecule is selected from the group consisting of: CD80; CD83; CD54; CD32; 4-1BBL; and CD70.