Li Xiu-Zhen, Wang Xiao-Hu, Xia Li-Jun, Weng Ya-Biao, Hernandez Jorge A, Tu Li-Qing, Li Lu-Tao, Li Shou-Jun, Yuan Zi-Guo
College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, PR China.
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases, Guangzhou, Guangdong Province, 510642, PR China.
BMC Infect Dis. 2015 Mar 4;15:114. doi: 10.1186/s12879-015-0815-1.
The use of recombinant viral vectors expressing T. gondii antigens is a safe and efficient approach to induce immune responses against the parasite, as well as a valuable tool for vaccine development. We have previously prolonged the survival time of mice challenged with the RH strain of T. gondii by immunizing the mice with a eukaryotic vector expressing the protein ROP18 of T. gondii. We are now looking for ways to improve this vaccination strategy and enhance protection.
In this study, we constructed and characterized a novel recombinant canine adenovirus type 2 expressing ROP18 (CAV-2-ROP18) of T. gondii by cytopathic effect (CPE) and indirect immunofluorescence assay (IFA) following transfection into MDCK cells. Intramuscular immunization of Kunming mice with CAV-2-ROP18 was carried out to evaluate humoral and cellular immune responses.
The vaccination of experimental mice with CAV-2-ROP18 elicited antibody production against ROP18, including high levels of a mixed IgG1/IgG2a and significant production of IFN-γ or IL-2, and displayed a significant bias towards a helper T cell type 1 (Th1) profile. Furthermore, the presence of T. gondii-specific IFN-γ-production and TNF-α-production T cells was elicited in both CD4+ and CD8+ T cell compartments. Significantly higher survival rates (40%) occurred in the experimental group, and a reduction in brain cyst burden was detected in vaccinated mice.
These results demonstrate the potential use of a CAV vector harboring the ROP18 gene in the development of a vaccine against acute and chronic toxoplasmosis.
使用表达弓形虫抗原的重组病毒载体是诱导针对该寄生虫的免疫反应的一种安全有效的方法,也是疫苗开发的一种有价值的工具。我们之前通过用表达弓形虫蛋白ROP18的真核载体免疫小鼠,延长了受弓形虫RH株攻击的小鼠的存活时间。我们现在正在寻找改进这种疫苗接种策略并增强保护作用的方法。
在本研究中,我们通过细胞病变效应(CPE)和间接免疫荧光法(IFA)构建并鉴定了一种新型的表达弓形虫ROP18的重组犬2型腺病毒(CAV-2-ROP18),将其转染到MDCK细胞中。用CAV-2-ROP18对昆明小鼠进行肌肉免疫,以评估体液免疫和细胞免疫反应。
用CAV-2-ROP18对实验小鼠进行疫苗接种引发了针对ROP18的抗体产生,包括高水平的混合IgG1/IgG2a以及显著的IFN-γ或IL-2产生,并且显示出对1型辅助性T细胞(Th1)谱的显著偏向。此外,在CD4+和CD8+ T细胞区室中均引发了弓形虫特异性IFN-γ产生和TNF-α产生的T细胞。实验组的存活率显著更高(40%),并且在接种疫苗的小鼠中检测到脑囊肿负担减轻。
这些结果证明了携带ROP18基因的CAV载体在开发针对急性和慢性弓形虫病疫苗中的潜在用途。