Nucleic acid-based formulation of cytomegalovirus-vectored HIV vaccines

这笔赠款用于可扩展的、基于核酸的配方的翻译开发 巨细胞病毒载体疫苗可以在没有冷链的情况下分发。HIV候选病毒 使用巨细胞病毒(CMV)作为递送载体和免疫调节佐剂的疫苗显示 不同寻常的承诺。例如，在我们的初步工作中，一种恒河猴巨细胞病毒载体SIV疫苗 缺乏病毒IL-10基因(RhCMVdIL10-SIVgag)保护了4/6的婴儿免受SIV感染。Tendel疗法 公司正在授权以这些第二代CMV载体疫苗为中心的技术组合。 以巨细胞病毒为基础的疫苗的制造和分销面临着艰巨的挑战：(1)在 培养速度明显慢于其他疫苗载体；(Ii)CMV在以下情况下经历快速遗传变化 在培养中扩增；(Iii)病毒被包裹，因此很难从细胞和病毒来源的脂质中分离出来 尺寸相似的双分子膜；(Iv)非均相颗粒的极端浓缩方法尚不清楚； 以及(V)配送需要冷链。 为了消除这些问题，Tendel还授权使用纯化的CMV运送疫苗的技术 基因组在大肠杆菌中繁殖。这项技术允许在引入基因后有效地拯救基因组 哺乳动物细胞。同样有效的活体救援应该会产生相当于 由常规接种病毒粒子引起的感染。 我们假设病毒IL-10缺陷的巨细胞病毒载体疫苗基因组(DNA)激发免疫反应 这与封装的活疫苗刺激的保护性反应没有区别。 目的1.评估在将疫苗基因组交付给 猕猴。CMV载体疫苗作为病毒粒子首先在当地复制，导致炎症细胞涌入， 然后系统地，导致病毒基因在远处组织中的表达。我们的假设预测 成功挽救疫苗基因组应该会导致同样的事件。 目的2.测试接种核酸疫苗的先天免疫反应和获得性免疫反应是否具有可比性 与先前观察到的保护性抗SIV反应有关。先前的研究表明，保护 抗SIV与特异性免疫反应有关，尤其是Mamu-E限制性CD8+T细胞 回应。 这些创新的第一阶段实验将足以建立技术优势和-鉴于 已证明对CMV载体疫苗的商业兴趣--Tendel方法的商业潜力。阶段 II实验将在这项工作的基础上进行，以(I)证明接种核酸疫苗的猕猴 保护免受SIV挑战(II)继续发展复制缺陷的HCMV-HIV Gag和Env 疫苗。

This grant is for translational development of a scalable, nucleic acid-based formulation of cytomegalovirus-vectored vaccines that can be distributed without a cold chain. HIV candidate vaccines that use cytomegalovirus (CMV) as delivery vector and immunomodulatory adjuvant have shown extraordinary promise. In our preliminary work, for example, a rhesus cytomegalovirus-vectored SIV vaccine lacking the viral IL-10 gene (RhCMVdIL10-SIVgag) protected 4/6 infants from SIV infection. Tendel Therapies Inc. is licensing a portfolio of technology centered on these second-generation, CMV-vectored vaccines. Manufacturing and distribution of CMV-based vaccines present daunting challenges: (i) replication of CMV in culture is markedly slower than that of other vaccine vectors; (ii) CMV undergoes rapid genetic change when amplified in culture; (iii) the virus is enveloped and thus difficult to separate from cell- and virus-derived lipid bilayers of a similar size; (iv) methods for extreme concentration of the heterogenous particles are unknown; and (v) a cold chain is required for distribution. To eliminate these problems, Tendel is also licensing technology for vaccine delivery using purified CMV genomes propagated in E. coli. The technology permits efficient “rescue” of the genomes after introduction to mammalian cells. Similarly efficient rescue in vivo should lead to immune responses that are equivalent to those provoked by conventional vaccination with virions. We hypothesize that viral IL-10-deficient, CMV-vectored vaccine genomes (DNA) provoke immune responses that are indistinguishable from the protective responses stimulated by encapsidated live vaccine. Aim 1. Assess antigen expression and vaccine vector replication after delivery of vaccine genomes to macaques. CMV-vectored vaccines given as virions first replicate locally, leading to inflammatory cell influx, and then systemically, leading to viral gene expression in distant tissues. Our hypothesis predicts that successful rescue of vaccine genomes should lead to the same events. Aim 2. Test if innate and adaptive immune responses to vaccination with nucleic acid are comparable to protective anti-SIV responses observed previously. Previous studies have shown that protection against SIV is associated with specific immune responses, particularly Mamu-E-restricted CD8+ T cell responses. These innovative Phase I experiments will be sufficient to establish both the technical merit and—in light of the proven commercial interest in CMV-vectored vaccines—the commercial potential of Tendel's approach. Phase II experiments will build on this work to (i) demonstrate that macaques vaccinated with nucleic acid are protected against SIV challenge (ii) continue development of replication-defective HCMV-HIV Gag and Env vaccines.