Zhao Zirun, Sun Ying, Li Mingchun, Yu Qilin
Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, College of Life Sciences, Nankai University, Tianjin 300071, China.
Research Center for Infectious Diseases, Nankai University, Tianjin 300350, China.
Vaccines (Basel). 2023 Sep 25;11(10):1521. doi: 10.3390/vaccines11101521.
The development of efficient fungal vaccines is urgent for preventing life-threatening systemic fungal infections. In this study, we prepared a synthetic, cell-based fungal vaccine for preventing systemic fungal infections using synthetic biology techniques. The synthetic cell EmEAP1 was constructed by transforming the chassis using a de novo synthetic fragment encoding the protein mChEap1 that was composed of the OmpA peptide, the fluorescence protein mCherry, the adhesin Eap1, and the C-terminally transmembrane region. The EmEAP1 cells highly exposed the mChEap1 on the cell surface under IPTG induction. The fungal vaccine was then prepared by mixing the EmEAP1 cells with aluminum hydroxide gel and CpG. Fluorescence quantification revealed that the fungal vaccine was stable even after 112 days of storage. After immunization in mice, the vaccine resided in the lymph nodes, inducing the recruitment of CD11c dendritic cells. Moreover, the vaccine strongly activated the CD4 T splenocytes and elicited high levels of anti-Eap1 IgG. By the prime-boost immunization, the vaccine prolonged the survival time of the mice infected by the cells and attenuated fungal colonization together with inflammation in the kidneys. This study sheds light on the development of synthetic biology-based fungal vaccines for the prevention of life-threatening fungal infections.
开发高效的真菌疫苗对于预防危及生命的系统性真菌感染至关重要。在本研究中，我们使用合成生物学技术制备了一种基于细胞的合成真菌疫苗，用于预防系统性真菌感染。合成细胞EmEAP1是通过用编码由OmpA肽、荧光蛋白mCherry、粘附素Eap1和C末端跨膜区域组成的蛋白质mChEap1的从头合成片段转化底盘而构建的。在IPTG诱导下，EmEAP1细胞在细胞表面高度暴露mChEap1。然后通过将EmEAP1细胞与氢氧化铝凝胶和CpG混合制备真菌疫苗。荧光定量显示，即使在储存112天后，真菌疫苗仍然稳定。在小鼠中免疫后，疫苗驻留在淋巴结中，诱导CD11c树突状细胞的募集。此外，该疫苗强烈激活CD4 T脾细胞并引发高水平的抗Eap1 IgG。通过初免-加强免疫，该疫苗延长了受细胞感染小鼠的存活时间，并减轻了肾脏中的真菌定植和炎症。这项研究为开发基于合成生物学的真菌疫苗以预防危及生命的真菌感染提供了思路。
