弓形虫ROP12核酸疫苗对小鼠弓形虫感染度的影响  

Toxoplasma gondii infectivity of a Toxoplasma ROP12 nucleic acid vaccine in mice

作　　者：王春苗[1] 姜文静[1] 郭玲玲[1] 张晓磊[1] 张进顺[1] 卢致民[1] 贾晓晖[1] WANG Chun-miao;JIANG Wen-jing;GUO Lin-lin;ZHANG Xiao-lei;ZHANG Jin-shun;LU Zhi-min;JIA Xiao-hui(Institute of Pathogen Biology ancl Immunology,Hebei North University,Zharigjiakou,Hebei 075000)

机构地区：[1]河北北方学院病原生物与免疫学研究所,河北张家口075000

出　　处：《中国病原生物学杂志》2021年第7期774-777,共4页Journal of Pathogen Biology

摘　　要：目的探讨弓形虫ROP12核酸疫苗对小鼠弓形虫感染度的影响。方法提取弓形虫RH株速殖子总RNA,根据ROP12基因全长编码序列(登录号为DQ096559)的开放阅读框设计引物并进行逆转录PCR(RT-PCR),扩增目的片段ROP12基因克隆至真核表达质粒pVAX1上,构建重组表达质粒pVAX1-ROP12,并进行酶切和测序鉴定。将小鼠随机分为pVAX1-ROP12组、pVAX1组及空白对照组,分别用相应接种物(空白对照不作处理)免疫小鼠3次,每次间隔2周,末次免疫2周后各组小鼠腹腔攻击感染RH株弓形虫速殖子1×10^(3)个/只,定时解剖小鼠,取组织印片并染色,观察各器官的虫体感染度。结果 RT-PCR扩增产物约为711 bp,菌落PCR及双酶切结果正确,序列测序结果与GenBank上报告的ROP12序列相似度为99.9%。用构建的弓形虫ROP12基因真核表达重组质粒pVAX1-ROP12免疫小鼠后,取弓形虫进行攻击感染,相同时间点小鼠各器官的感染度较空质粒对照和空白对照显著降低(均P<0.05)。结论 ROP12核酸疫苗免疫对弓形虫在小鼠各组织器官的感染度有显著的抑制作用,这为弓形虫疫苗提供理论基础。Objective To investigate the Toxoplasma gondii infectivity of a Toxoplasma ROP12 nucleic acid vaccine in mice. Methods Total RNA of T. gondii was extracted from RH strain tachyzoites, and primers were designed in accordance with the open reading frame of the ROP12 gene encoding sequence(registration No.DQ096559). The ROP12 gene was amplified with reverse transcription PCR(RT-PCR) and cloned into the eukaryotic expression plasmid pVAX1 to construct the recombinant plasmid pVAX1-ROP12. The gene was then identified using enzyme digestion and sequencing. Mice were randomly divided into a pVAX1-ROP12 group, a pVAX1 group, and a blank control group. All groups were immunized with the corresponding inoculum(the blank control group was not treated) 3 times at 2-week intervals. The mice in each group were intraperitoneal attacked and infected with 1×10^(3) T. gondii RH strain tachyzoites 2 wks after the last injection, and dissected regularly to observe the infectivity of each organ by tissue printing and staining. Results RT-PCR indicated that a fragment of about 711 bp was amplified from T. gondii RH strain tachyzoites. Colony-PCR, double enzyme digestion, and sequencing indicated that the recombinant plasmid pVAX1-ROP12 was successfully constructed. Mice infected with T. gondii were immunized using the recombinant plasmid pVAX1-ROP12, pVAX1, and the blank control. Statistical analysis indicated that the infectivity of each organ in the pVAX1-ROP12 group was significantly lower than that in the pVAX1 group and the blank control group at the same time point(P<0.05 for both). Conclusion Results indicated that immunization with a ROP12 nucleic acid vaccine inhibits the infectivity of T. gondii in various tissues and organs of mice. This finding will provide a theoretical basis for the further study of Toxoplasma vaccines.

关 键 词：刚地弓形虫 棒状体蛋白ROP12 核酸疫苗 弓形虫感染度 

分 类 号：R382.5[医药卫生—医学寄生虫学]