African horse sickness (AHS) is a devastating viral disease affecting equines and has resulted in many disastrous epizootics. To date, no successful therapeutic treatment exists for AHS, and commercially used live-attenuated vaccines have various undesirable side effects. Previous studies have shown that mice inoculated with insoluble African horse sickness virus (AHSV) VP7 crystals are protected from live challenge with a lethal dose of AHSV. This study investigates the humoral and cell-mediated immune responses in guinea-pigs to a safer monovalent vaccine alternative based on AHSV-5 VP7 quasi-crystals produced in plants. Guinea-pigs received prime- and boost-inoculations of between 10 and 50 μg of purified plant-produced AHSV VP7. Western immunoblot analysis of the humoral response showed stimulation of high titres of anti-VP7 antibodies 28 days after the boost-inoculation in sera from three of the five experimental animals. In addition, RNA-seq transcriptome profiling of guinea-pig spleen-derived RNA highlighted thirty significantly (q ≤ 0.05) differentially expressed genes involved in innate and adaptive immunity. Differential expression of genes involved in Th1, Th2 and Th17 cell differentiation suggest a cell-mediated immune response to AHSV-5 VP7. Upregulation of several important cytokines and cytokine receptors were noted, including TNFSF14, CX3CR1, IFNLR1 and IL17RA. Upregulation of IL17RA suggests a Th17 response which has been reported as a key component in AHSV immunity. While further investigation is needed to validate these findings, these results suggest that AHSV-5 VP7 quasi-crystals produced in N. benthamiana are immunogenic and induce both humoral and cell-mediated responses.

非洲马病 (AHS) 是一种影响马科动物的破坏性病毒性疾病，并导致了许​​多灾难性的动物流行病。迄今为止，AHS 尚无成功的治疗方法，并且商业上使用的减毒活疫苗具有各种不良副作用。先前的研究表明，接种不溶性非洲马病病毒 (AHSV) VP7 晶体的小鼠可以免受致命剂量 AHSV 的活体攻击。本研究调查了豚鼠对基于植物产生的 AHSV-5 VP7 准晶体的更安全的单价疫苗替代品的体液和细胞介导的免疫反应。豚鼠接受了 10 至 50 μg 纯化的植物产生的 AHSV VP7 的初免和加强接种。体液反应的蛋白质免疫印迹分析显示，在来自五只实验动物中的三只的血清中加强接种后 28 天，刺激了高滴度的抗 VP7 抗体。此外，豚鼠脾源性 RNA 的 RNA-seq 转录组分析突出了 30 个显着（q ≤ 0.05）与先天性和适应性免疫有关的差异表达基因。参与 Th1、Th2 和 Th17 细胞分化的基因的差异表达表明细胞介导的对 AHSV-5 VP7 的免疫反应。注意到几种重要的细胞因子和细胞因子受体的上调，包括 TNFSF14、CX3CR1、IFNLR1 和 IL17RA。IL17RA 的上调表明 Th17 反应已被报道为 AHSV 免疫的关键组成部分。虽然需要进一步调查来验证这些发现，本氏烟草具有免疫原性并诱导体液和细胞介导的反应。