Novavax AB, Kungsgatan 109, SE75653 Uppsala, Sweden.
Novavax, Inc., 22 Firstfield Road, Gaithersburg, MD 20878, United States.
Vaccine. 2016 Apr 7;34(16):1927-35. doi: 10.1016/j.vaccine.2016.02.033. Epub 2016 Feb 24.
Ebola virus (EBOV) causes severe hemorrhagic fever for which there is no approved treatment or preventive vaccine. Immunological correlates of protective immunity against EBOV disease are not well understood. However, non-human primate studies have associated protection of experimental vaccines with binding and neutralizing antibodies to the EBOV glycoprotein (GP) as well as EBOV GP-specific CD4(+) and CD8(+) T cells. In this report a full length, unmodified Zaire EBOV GP gene from the 2014 EBOV Makona strain (EBOV/Mak) was cloned into a baculovirus vector. Recombinant EBOV/Mak GP was produced in Sf9 insect cells as glycosylated trimers and, when purified, formed spherical 30-40 nm particles. In mice, EBOV/Mak GP co-administered with the saponin adjuvant Matrix-M was significantly more immunogenic, as measured by virus neutralization titers and anti-EBOV/Mak GP IgG as compared to immunization with AlPO4 adjuvanted or non-adjuvanted EBOV/Mak GP. Similarly, antigen specific T cells secreting IFN-γ were induced most prominently by EBOV/Mak GP with Matrix-M. Matrix-M also enhanced the frequency of antigen-specific germinal center B cells and follicular helper T (TFH) cells in the spleen in a dose-dependent manner. Immunization with EBOV/Mak GP with Matrix-M was 100% protective in a lethal viral challenge murine model; whereas no protection was observed with the AlPO4 adjuvant and only 10% (1/10) mice were protected in the EBOV/Mak GP antigen alone group. Matrix-M adjuvanted vaccine induced a rapid onset of specific IgG and neutralizing antibodies, increased frequency of multifunctional CD4+ and CD8(+) T cells, specific TFH cells, germinal center B cells, and persistence of EBOV GP-specific plasma B cells in the bone marrow. Taken together, the addition of Matrix-M adjuvant to the EBOV/Mak GP nanoparticles enhanced both B and T-cell immune stimulation which may be critical for an Ebola subunit vaccine with broad and long lasting protective immunity.
埃博拉病毒(EBOV)可引起严重的出血热,目前尚无批准的治疗或预防疫苗。针对 EBOV 疾病的保护性免疫的免疫学相关性尚未得到很好的理解。然而,非人类灵长类动物研究将实验疫苗的保护与针对 EBOV 糖蛋白(GP)的结合和中和抗体以及 EBOV GP 特异性 CD4(+)和 CD8(+)T 细胞联系起来。在本报告中,从 2014 年 EBOV Makona 株(EBOV/Mak)克隆了全长、未经修饰的扎伊尔 EBOV GP 基因到杆状病毒载体中。重组 EBOV/Mak GP 在 Sf9 昆虫细胞中作为糖基化三聚体产生,纯化后形成球形 30-40nm 颗粒。在小鼠中,EBOV/Mak GP 与皂苷佐剂 Matrix-M 联合使用时,与 AlPO4 佐剂或非佐剂的 EBOV/Mak GP 免疫相比,其免疫原性显著更高,表现为病毒中和滴度和抗 EBOV/Mak GP IgG。同样,通过 EBOV/Mak GP 与 Matrix-M 诱导产生 IFN-γ的抗原特异性 T 细胞最为显著。Matrix-M 还以剂量依赖性方式增强了脾脏中抗原特异性生发中心 B 细胞和滤泡辅助 T(TFH)细胞的频率。在致命病毒挑战的小鼠模型中,EBOV/Mak GP 与 Matrix-M 免疫接种 100%具有保护作用;而 AlPO4 佐剂无保护作用,在 EBOV/Mak GP 抗原单独组中只有 10%(1/10)的小鼠具有保护作用。Matrix-M 佐剂疫苗诱导特异性 IgG 和中和抗体的快速产生,增加了多功能 CD4+和 CD8(+)T 细胞、特异性 TFH 细胞、生发中心 B 细胞以及骨髓中 EBOV GP 特异性浆细胞的频率。综上所述,Matrix-M 佐剂的加入增强了 EBOV/Mak GP 纳米颗粒的 B 和 T 细胞免疫刺激,这对于具有广泛和持久保护免疫的 Ebola 亚单位疫苗可能是至关重要的。
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