Table3_Functional genomic analysis of the 68-1 RhCMV-Mycobacteria tuberculosis vaccine reveals an IL-15 response signature that is conserved with vector attenuation.xlsx

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb) is a deadly infectious disease having a major impact on global health. Using the CMV vector for development of novel vaccines is a promising new strategy that elicits strong and durable, high frequency memory T cell responses against heterologous immunogens. We conducted functional transcriptomic analysis of whole blood samples collected from cohorts of rhesus (Rh) macaques that were administered RhCMV/TB vector using a prime-boost strategy. Two modified CMV vectors were used in this study, including 68-1 RhCMV/TB-6Ag (encoding 6 Mtb protein immunogens, including Ag85A, ESAT-6, Rv3407, Rv2626, Rpf A, and Rpf D) and its attenuated variant, 68-1 RhCMV/Δpp71-TB-6Ag (a cell-to-cell spread-deficient vaccine vector lacking the Rh110 gene encoding the pp71 tegument protein). Bulk mRNA sequencing, differential gene expression, and functional enrichment analyses showed that these RhCMV/TB vaccines induce the innate and adaptive immune responses with specific transcriptomic signatures, including the IL-15-induced protective gene signature previously defined to be linked with protection against simian immunodeficiency virus (SIV) by the 68-1 RhCMV/SIV vaccine. While both vectors exhibited a transcriptomic response of the IL-15 protective signature in whole blood, we show that lack of pp71 does not maintain induction of the protective signature for the full duration of the study compared to the parental non-attenuated vector. Our observations indicate that RhCMV vector vaccines induce a transcriptomic response in whole blood that include a conserved IL-15 signature of which vector-encoded pp71 is an important component of response durability that upon future Mtb challenge may define specific vaccine protection outcomes against Mtb infection.

结核病（TB），由分枝杆菌属结核分支杆菌（Mycobacterium tuberculosis，Mtb）引起，是一种对全球公共卫生产生重大影响的致命性传染病。利用CMV病毒载体开发新型疫苗是一种前景广阔的新策略，该策略能够引发针对异源免疫原的强大且持久的、高频记忆T细胞反应。我们采用原-加强策略，对接受RhCMV/TB病毒载体接种的恒河猴（Rh）群体进行了全血样本的功能性转录组学分析。本研究中使用了两种改造后的CMV病毒载体，包括编码6种Mtb蛋白免疫原（包括Ag85A、ESAT-6、Rv3407、Rv2626、Rpf A和Rpf D）的68-1 RhCMV/TB-6Ag及其减毒变种，68-1 RhCMV/Δpp71-TB-6Ag（一种细胞间传播缺陷型疫苗载体，缺乏编码pp71衣壳蛋白的Rh110基因）。大量mRNA测序、差异基因表达和功能富集分析表明，这些RhCMV/TB疫苗能够引发先天性和适应性免疫反应，并具有特定的转录组学特征，包括先前由68-1 RhCMV/SIV疫苗定义的与保护对猴免疫缺陷病毒（SIV）的IL-15诱导的保护基因特征。尽管两种载体在整血中均表现出IL-15保护特征的转录组反应，但我们发现，与亲本非减毒载体相比，缺乏pp71不能在整个研究期间维持保护特征的诱导。我们的观察结果表明，RhCMV病毒载体疫苗能够引发整血中的转录组反应，其中包括一个保守的IL-15特征，其中载体编码的pp71是反应持久性的重要组成部分，在未来的Mtb挑战中，它可能定义针对Mtb感染的特定疫苗保护结果。