Calazans Alexandre, Boggiano Cesar, Lindsay Ross
Design and Development Laboratory, International AIDS Vaccine Initiative, Brooklyn, NY, United States of America.
Center for Technological Development in Health, Oswaldo Cruz, Foundation, Rio de Janeiro, Brazil.
PLoS One. 2017 Aug 24;12(8):e0183803. doi: 10.1371/journal.pone.0183803. eCollection 2017.
We developed a DNA vaccine that induces the formation of a VLP in vivo. This VLP was designed to elicit neutralizing antibodies, to induce better T-cell responses and to activate the innate immune system. Overall, 5 groups of 10 mice were electroporated with the following constructs: pVLP-LTR-GagPro [full], pVLP-GagPro [VLP wihout RNA], pVLP-LTR-Gag [VLP immature], pVLP-Gag and pVLP-EnvBG505 [regular DNA vaccine] and a mock group. We performed ICS on the mouse spleens and performed ELISA for ENV antibodies and a Luminex assay for inflammatory cytokines. The VLP showed good binding to the neutralizing antibodies. The percentage of CD4 cells producing cytokines was 0.1% [IFNg], 0.15%[IL-2] and 0.2% [TNFa] for the construct pVLP-LTR-GagPro. The percentage of CD8 cells producing cytokines was 0.3%[IFNg], 0.2%[IL-2] and 0.25%[TNFa]. All pVLP constructs induced more antibodies for EnvBG505 than the regular DNA vaccine Env. The pVLP-LTR-GagPro induced more IL-1B than the other constructs 24 hours post-vaccination.
我们研发了一种能在体内诱导形成病毒样颗粒（VLP）的DNA疫苗。这种VLP旨在引发中和抗体，诱导更好的T细胞反应并激活先天免疫系统。总体而言，将5组每组10只小鼠用以下构建体进行电穿孔处理：pVLP-LTR-GagPro[完整]、pVLP-GagPro[不含RNA的VLP]、pVLP-LTR-Gag[未成熟VLP]、pVLP-Gag和pVLP-EnvBG505[常规DNA疫苗]以及一个模拟组。我们对小鼠脾脏进行了细胞内细胞因子染色（ICS），并对ENV抗体进行了酶联免疫吸附测定（ELISA），对炎性细胞因子进行了Luminex检测。该VLP与中和抗体表现出良好的结合。对于构建体pVLP-LTR-GagPro，产生细胞因子的CD4细胞百分比为0.1%[干扰素γ（IFNg）]、0.15%[白细胞介素-2（IL-2）]和0.2%[肿瘤坏死因子α（TNFa）]。产生细胞因子的CD8细胞百分比为0.3%[IFNg]、0.2%[IL-2]和0.25%[TNFa]。所有pVLP构建体诱导产生的针对EnvBG505的抗体都比常规DNA疫苗Env更多。接种疫苗24小时后，pVLP-LTR-GagPro诱导产生的白细胞介素-1β（IL-1B）比其他构建体更多。
