Identification of preexisting adaptive immunity to Cas9 proteins in humans
Identification of preexisting adaptive immunity to Cas9 proteins in humans
复制标题
DOI：
10.1038/s41591-018-0326-x
复制
发表时间：
2019-02-01
期刊：
影响因子：
82.9
通讯作者：
Porteus, Matthew H.
中科院分区：
文献类型：
--
作者：
Charlesworth, Carsten T.;Deshpande, Priyanka S.;Porteus, Matthew H.
CRISPR-Cas9系统是一种强大的基因组编辑工具，可以精确修饰特定的DNA序列。人们正在努力使用CRISPR-Cas9系统来治疗纠正人类遗传疾病（1-6）。最广泛使用的Cas9的直系同源物来源于金黄色葡萄球菌和酿脓链球菌（5，7）。鉴于这两种细菌物种以高频率感染人类群体（8，9），我们假设人类可能对源自这些细菌物种的Cas9直系同源物SaCas 9（S. aureus）和SpCas 9（S.化脓性）。通过使用酶联免疫吸附测定探测人血清中抗Cas9抗体的存在，我们分别在78%和58%的供体中检测到针对SaCas 9和SpCas 9的抗体。我们还在78%的供体中发现了抗SaCas 9 T细胞，在67%的供体中发现了抗SpCas 9 T细胞，这证明了针对两种直系同源物的抗原特异性T细胞的高流行率。我们通过证明在分离、扩增和抗原再刺激后的Cas9特异性细胞因子应答来证实这些T细胞是Cas9特异性的。总之，这些数据表明，在人类中存在对Cas9的预先存在的体液和细胞介导的适应性免疫应答，这一发现应该在CRISPR-Cas9系统走向临床试验时加以考虑。
The CRISPR-Cas9 system is a powerful tool for genome editing, which allows the precise modification of specific DNA sequences. Many efforts are underway to use the CRISPR-Cas9 system to therapeutically correct human genetic diseases(1-6). The most widely used orthologs of Cas9 are derived from Staphylococcus aureus and Streptococcus pyogenes(5,7). Given that these two bacterial species infect the human population at high frequencies(8,9), we hypothesized that humans may harbor preexisting adaptive immune responses to the Cas9 orthologs derived from these bacterial species, SaCas9 (S. aureus) and SpCas9 (S. pyogenes). By probing human serum for the presence of anti-Cas9 antibodies using an enzyme-linked immunosorbent assay, we detected antibodies against both SaCas9 and SpCas9 in 78% and 58% of donors, respectively. We also found anti-SaCas9 T cells in 78% and anti-SpCas9 T cells in 67% of donors, which demonstrates a high prevalence of antigen-specific T cells against both orthologs. We confirmed that these T cells were Cas9-specific by demonstrating a Cas9-specific cytokine response following isolation, expansion, and antigen restimulation. Together, these data demonstrate that there are preexisting humoral and cell-mediated adaptive immune responses to Cas9 in humans, a finding that should be taken into account as the CRISPR-Cas9 system moves toward clinical trials.