Genetic Ablation of HLA Class I, Class II, and the T-cell Receptor Enables Allogeneic T Cells to Be Used for Adoptive T-cell Therapy
Genetic Ablation of HLA Class I, Class II, and the T-cell Receptor Enables Allogeneic T Cells to Be Used for Adoptive T-cell Therapy
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DOI：
10.1158/2326-6066.cir-18-0508
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发表时间：
2020-07-01
影响因子：
10.1
通讯作者：
Hirano, Naoto
中科院分区：
文献类型：
--
作者：
Kagoya, Yuki;Guo, Tingxi;Hirano, Naoto
过继免疫治疗可以在某些癌症中产生持续的治疗效果。抗肿瘤T细胞移植通常是由自体T细胞在体外单独制备的，这需要高强度的工作量和增加的成本。产生的T细胞的质量也可能是不同的，这影响了治疗的抗肿瘤效果和毒性。如果同种异体T细胞反应得到有效控制，来自第三方捐赠者的抗肿瘤T细胞移植物的标准化生产将使这种方式得到广泛应用。在这里，我们通过Cas9/sgRNA核糖核蛋白电穿孔同时敲除B2M、CIITA和TRAC基因，产生了HLAI类、HLAII类和T细胞受体(TCR)三重敲除(TKO)T细胞。尽管自然杀伤细胞针对的是人类白细胞抗原缺陷的T细胞，但在免疫缺陷小鼠中，在同种异体外周血单核细胞(PBMC)存在的情况下，它们的存活率要好于人类白细胞抗原充足的T细胞。用CD19嵌合抗原受体(CAR)转导TKO CAR-T细胞并经肿瘤细胞刺激后，TKO CAR-T细胞与同种异体PBMC共同培养时，其存活率明显高于TRAC和B2M双基因敲除T细胞。CD19 TKO CAR-T细胞不能诱发移植物抗宿主病，但保留了抗肿瘤反应。这些结果证明了HLAI类、HLAII类和TCR缺失在使同种异体来源的T细胞能够用于现成过继免疫治疗方面的好处。
Adoptive immunotherapy can induce sustained therapeutic effects in some cancers. Antitumor T-cell grafts are often individually prepared in vitro from autologous T cells, which requires an intensive workload and increased costs. The quality of the generated T cells can also be variable, which affects the therapy's antitumor efficacy and toxicity. Standardized production of antitumor T-cell grafts from third-party donors will enable widespread use of this modality if allogeneic T-cell responses are effectively controlled. Here, we generated HLA class I, HLA class II, and T-cell receptor (TCR) triple-knockout (tKO) T cells by simultaneous knockout of the B2M, CIITA, and TRAC genes through Cas9/sgRNA ribonucleoprotein electroporation. Although HLA-deficient T cells were targeted by natural killer cells, they persisted better than HLA-sufficient T cells in the presence of allogeneic peripheral blood mononuclear cells (PBMC) in immunodeficient mice. When transduced with a CD19 chimeric antigen receptor (CAR) and stimulated by tumor cells, tKO CAR-T cells persisted better when cultured with allogeneic PBMCs compared with TRAC and B2M double-knockout T cells. The CD19 tKO CAR-T cells did not induce graft-versus-host disease but retained antitumor responses. These results demonstrated the benefit of HLA class I, HLA class II, and TCR deletion in enabling allogeneic-sourced T cells to be used for off-the-shelf adoptive immunotherapy.