Institute of Zoonosis, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, China; Jilin Provincial Animal Disease Control Center, 4510 Xi'an Road, Changchun 130062</postal>, China.
Vaccine. 2013 Dec 9;31(51):6065-71. doi: 10.1016/j.vaccine.2013.10.015. Epub 2013 Oct 29.
The investigation of Toxoplasma gondii virulence factors can elucidate the immunopathology of T. gondii infection and identify potential candidates for effective human vaccines. The adjuvant is an important component of an effective vaccine. In this study, attenuated Mycobacterium bovis was used as a live vaccine vector with both antigen and adjuvant characteristics. Following amplification of the T. gondii cyclophilin gene, the shuttle expression plasmid pMV261-TgCyP and integrative expression plasmid pMV361-TgCyP were constructed, and their expression was stimulated after transfection into BCG. Both recombinant plasmids were highly immunogenic. Greater proliferation of CD4(+) and CD8(+) T cells was observed in the rBCG-vaccinated groups compared to the control groups. The levels of Th1-type IFN-γ, IL-2 and IL-12 were significantly increased following immunisation with the rBCG vaccines via the i.v. or oral route, which indicated that catalytic activity against T. gondii infection was generated in the mice. rBCGpMV361-TgCyP i.v. inoculation resulted in a higher protection efficiency, as demonstrated by the increased survival time and survival rate (17%) of BALB/c mice. The present study demonstrates that a BCG vector expressing a target antigen, TgCyP, represent an alternative system for the production of effective vaccines to prevent toxoplasmosis.
弓形虫毒力因子的研究可以阐明弓形虫感染的免疫病理学,并确定有效的人类疫苗的潜在候选物。佐剂是有效疫苗的重要组成部分。在这项研究中,减毒牛分枝杆菌被用作具有抗原和佐剂特性的活疫苗载体。扩增弓形虫亲环素基因后,构建了穿梭表达质粒 pMV261-TgCyP 和整合表达质粒 pMV361-TgCyP,并在转染 BCG 后刺激其表达。两种重组质粒均具有高度的免疫原性。与对照组相比,rBCG 疫苗接种组 CD4(+)和 CD8(+)T 细胞的增殖更为明显。通过静脉内或口服途径用 rBCG 疫苗免疫后,Th1 型 IFN-γ、IL-2 和 IL-12 的水平显著升高,这表明在小鼠中产生了针对弓形虫感染的催化活性。rBCGpMV361-TgCyP 静脉内接种导致更高的保护效率,表现为 BALB/c 小鼠的存活时间和存活率(17%)增加。本研究表明,表达靶抗原 TgCyP 的 BCG 载体代表了预防弓形虫病的有效疫苗生产的替代系统。