Shimada Masaru, Wang Haibin, Ichino Motohide, Ura Takehiro, Mizuki Nobuhisa, Okuda Kenji
Department of Molecular Biodefense Research, Yokohama City University, Yokohama, 2360004, Japan.
BioRay Pharmaceutical Co., Ltd., Taizhou, Zhejiang, 318000, China.
Gene Ther. 2022 Nov;29(10-11):636-642. doi: 10.1038/s41434-021-00308-z. Epub 2022 Jan 6.
Previously, we developed a chimeric adenovirus type 5 with type 35 fiber (Ad5/35), which has high tropism to dendritic cells and low hepatoxicity. For further clinical use, we constructed two recombinant vectors expressing human immunodeficiency virus 1 (HIV-1) clade C gag (Ad5/35-Cgag and MVA-Cgag). The biodistribution of the two viral vectors in a mouse model and immunity in monkeys were assessed. The mice received a single intramuscular injection with the vectors alone. The gag gene in the tissues were periodically detected using a real-time quantitative polymerase chain reaction. The distribution of Ad5/35 was also detected using an in vivo imaging system, followed by luciferase-expressing Ad5/35 administration. We found that Ad5/35-Cgag DNA and luciferase activity were detectable until 8 weeks post-administration, whereas MVA-Cgag was undetectable 72 h post-administration. Furthermore, viral administration did not increase serum aspartate aminotransferase and alanine aminotransferase levels in either mouse or monkey models. Moreover, intramuscular administration of Ad5/35-Cgag induced the gag-specific antibody level and IFNγ-secreting PBMCs, the boost with MVA-Cgag further increased the responses and lasted more than 20 weeks from the initial administration. These data demonstrate that Ad5/35 and MVA vectors are safe for in vivo use, and prime-boost with Ad5/35-MVA vaccines is suitable for clinical use against HIV-1 clade C.
此前,我们构建了一种携带35型纤维的嵌合5型腺病毒(Ad5/35),其对树突状细胞具有高嗜性且肝毒性低。为了进一步用于临床,我们构建了两种表达人免疫缺陷病毒1型(HIV-1)C亚型gag的重组载体(Ad5/35-Cgag和MVA-Cgag)。评估了这两种病毒载体在小鼠模型中的生物分布以及在猴子中的免疫情况。小鼠单独接受一次肌肉注射载体。使用实时定量聚合酶链反应定期检测组织中的gag基因。在给予表达荧光素酶的Ad5/35后,还使用体内成像系统检测Ad5/35的分布。我们发现,直到给药后8周仍可检测到Ad5/35-Cgag DNA和荧光素酶活性,而MVA-Cgag在给药后72小时就无法检测到。此外,在小鼠或猴子模型中,病毒给药均未增加血清天冬氨酸转氨酶和丙氨酸转氨酶水平。而且,肌肉注射Ad5/35-Cgag可诱导gag特异性抗体水平和分泌IFNγ的外周血单核细胞,用MVA-Cgag加强免疫后进一步增强了反应,并且从初次给药起持续超过20周。这些数据表明,Ad5/35和MVA载体在体内使用是安全的,并且Ad5/35-MVA疫苗的初免-加强免疫方案适用于针对HIV-1 C亚型的临床应用。