Human Treg responses allow sustained recombinant adeno-associated virus-mediated transgene expression
Human Treg responses allow sustained recombinant adeno-associated virus-mediated transgene expression
复制标题
DOI：
10.1172/jci70314
复制
发表时间：
2013-12-01
影响因子：
15.9
通讯作者：
Flotte, Terence R.
中科院分区：
文献类型：
--
作者：
Mueller, Christian;Chulay, Jeffrey D.;Flotte, Terence R.
重组腺相关病毒（rAAV）载体已经显示出用于治疗几种疾病的前景;然而，已经提出免疫介导的转导细胞的消除限制并解释功效的丧失。为了确定rAAV载体表达是否可以长期持续，我们通过多次i.m.注射剂在所有受试者中均观察到M特异性AAT表达，呈剂量依赖性，并且在无免疫抑制的情况下持续超过1年。与3个月活检相比，1年时的肌肉活检具有持续的AAT表达和炎性细胞减少。来自肌肉活检的TCR V β区的深度测序证明了在载体施用后3个月出现的有限数量的T细胞克隆并持续1年。原位免疫表型分析揭示了大量的Treg群体。在含有AAT表达肌纤维的肌肉活检样品中。肌肉中约10%的所有T细胞是天然T细胞，其响应于AAV衣壳而被激活。这些结果表明，i.m. rAAV 1型-AAT（rAAV 1-AAT）的递送诱导T调节应答，其允许持续的转基因表达，并表明免疫调节治疗对于rAAV介导的基因治疗可能不是必需的。
Recombinant adeno-associated virus (rAAV) vectors have shown promise for the treatment of several diseases; however, immune-mediated elimination of transduced cells has been suggested to limit and account for a loss of efficacy. To determine whether rAAV vector expression can persist long term, we administered rAAV vectors expressing normal, M-type alpha-1 antitrypsin (M-AAT) to AAT-deficient subjects at various doses by multiple i.m. injections. M-specific AAT expression was observed in all subjects in a dose-dependent manner and was sustained for more than 1 year in the absence of immune suppression. Muscle biopsies at 1 year had sustained AAT expression and a reduction of inflammatory cells compared with 3 month biopsies. Deep sequencing of the TCR V beta region from muscle biopsies demonstrated a limited number of T cell clones that emerged at 3 months after vector administration and persisted for 1 year. In situ immunophenotyping revealed a substantial Treg population. in muscle biopsy samples containing AAT-expressing myofibers. Approximately 10% of all T cells in muscle were natural Tregs, which were activated in response to AAV capsid. These results suggest that i.m. delivery of rAAV type 1-AAT (rAAV1-AAT) induces a T regulatory response that allows ongoing transgene expression and indicates that immunomodulatory treatments may not be necessary for rAAV-mediated gene therapy.