VIRxSYS, Corporation, Gaithersburg, MD, USA.
J Gene Med. 2013 Feb;15(2):78-82. doi: 10.1002/jgm.2691.
Lentiviral vectors are being used with increasing frequency in human clinical trials. We were the first to use lentiviral vectors in clinical trials in 2003. Our lentiviral vector encoded a long RNA antisense sequence to the HIV-1 envelope and was used in an ex vivo autologous setting to provide viral load control in HIV-1 positive subjects failing anti-HIV therapy. A total of 65 subjects have been treated in Phase 1 and Phase 2 trials in six institutions.
Good manufacturing practices (GMP) lots of the lentiviral vector used in our clinical trials were assayed for the presence of replication competent lentivirus (RCL). RCL assays were conducted at two stages. The first testing was performed on samples collected immediately following bulk harvest of the GMP product lot and consisted of 1 × 10(8) cells used in production. RCL assays were also performed on aliquots of the final fill of the vector by the inoculation of at least 5% of the GMP final fill volume into C8166 cells, passaged for at least ten passages and tested for RCL by p24 enzyme-linked immunosorbent assay and vesicular stomatitis virus-G envelope DNA.
Following 263 infusions of autologous, transduced cells, no adverse events have been detected in these subjects, with some followed for more than 8 years following infusions. More than 4.3 × 10(12) VRX496 proviral copies were administered to these 65 subjects.
Data from this small population suggest that there is no apparent risk for serious adverse events with the use of lentiviral vectors.
慢病毒载体在人体临床试验中的使用频率越来越高。我们是在 2003 年首次在临床试验中使用慢病毒载体的。我们的慢病毒载体编码了一个长的 HIV-1 包膜 RNA 反义序列，用于体外自体环境中，为抗 HIV 治疗失败的 HIV-1 阳性患者提供病毒载量控制。共有 65 名受试者在六个机构的 1 期和 2 期临床试验中接受了治疗。
我们临床试验中使用的慢病毒载体的良好生产规范（GMP）批次用于检测复制能力强的慢病毒（RCL）的存在。RCL 检测分两个阶段进行。第一阶段的检测是在 GMP 产品批次的批量收获后立即进行的，包括用于生产的 1×10(8)个细胞。RCL 检测也在向量的最终填充等分样中进行，通过将至少 5%的 GMP 最终填充体积接种到 C8166 细胞中，传代至少十次，并通过 p24 酶联免疫吸附试验和水疱性口炎病毒-G 包膜 DNA 检测 RCL。
在 263 次自体转导细胞输注后，这些受试者没有检测到不良反应，其中一些受试者在输注后随访超过 8 年。这些 65 名受试者接受了超过 4.3×10(12)个 VRX496 前病毒拷贝。
来自这一小部分人群的数据表明，使用慢病毒载体没有明显的严重不良事件风险。
