Yu Guang-qing, Liu Wen-qi, Lei Jia-hui, Mo Hong-mei, Chen Yu-li, Li Yong-long
Department of Parasitology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2006 Feb 28;24(1):51-5.
To study the protective efficacy of co-immunization with Schistosoma japonicum glutathione S-transferase (Sj26) DNA and recombinant Sj26 protein (rSj26 GST) vaccine against Schistosoma japonicum in BALB/c mice.
The Sj26 gene was cloned into eukaryotic expression vector pEGFP-N3 with enhanced green fluorescence protein. The recombinant plasmid pEGFP-Sj26 was transfected into baby hamster kidney (BHK) cells, fluorescent microscope and Western blotting were employed to identify the expressed products. Each mouse in co-immunization group was primed with plasmid pEGFP-Sj26, boosted 2 weeks later and immunized with rSj26 GST 4 weeks later. While each mouse in pEGFP-Sj26 group and rSj26 GST group was primed and boosted with pEGFP-Sj26 or rSj26 GST independently. Two weeks after last immunization, each mouse was challenged with 40 +/- 1 cercariae of S. japonicum Chinese strain. At the 45th day post-infection, mice were sacrificed and the worms were perfused from portal vein and the number of worms and eggs in liver tissue were counted.
In BHK cells transfected with the recombinant plasmid pEGFP-Sj26, the expression of Sj26-EGFP fusion protein was confirmed by fluorescent microcopy and Western blotting. The worm reduction rate in co-immunized group was 50.8%, significantly higher than that in pEGFP-Sj26 group (28.0%, P < 0.01) and rSj26 GST group (25.5%, P < 0.01). Liver egg reduction rate in co-immunized group, pEGFP-Sj26 group and rSj26 GST group were 32.7%, 20.6% and 33.0% respectively. The number of eggs per female in liver of co-immunized mice and rSj26 GST group were significantly higher than that in control group (P < 0.01).
Compared to the immunization with pEGFP-Sj26 or rSj26 GST alone, the co-immunization with pEGFP-Sj26 and rSj26 GST can enhance protective efficacy in BALB/c mice.
研究日本血吸虫谷胱甘肽S-转移酶(Sj26)DNA与重组Sj26蛋白(rSj26 GST)联合免疫对BALB/c小鼠日本血吸虫感染的保护效果。
将Sj26基因克隆至带有增强型绿色荧光蛋白的真核表达载体pEGFP-N3中。将重组质粒pEGFP-Sj26转染至幼仓鼠肾(BHK)细胞,采用荧光显微镜和蛋白质印迹法鉴定表达产物。联合免疫组每只小鼠先用质粒pEGFP-Sj26免疫,2周后加强免疫,4周后用rSj26 GST免疫。而pEGFP-Sj26组和rSj26 GST组每只小鼠分别单独用pEGFP-Sj26或rSj26 GST进行初次免疫和加强免疫。末次免疫2周后,每只小鼠用40±1条日本血吸虫中国株尾蚴攻击感染。感染后第45天,处死小鼠,经门静脉灌注收集虫体,计数肝脏组织中的虫体和虫卵数量。
在转染重组质粒pEGFP-Sj26的BHK细胞中,荧光显微镜和蛋白质印迹法证实了Sj26-EGFP融合蛋白的表达。联合免疫组的减虫率为50.8%,显著高于pEGFP-Sj26组(28.0%,P<0.01)和rSj26 GST组(25.5%,P<0.01)。联合免疫组、pEGFP-Sj26组和rSj26 GST组的肝脏减卵率分别为32.7%、20.6%和33.0%。联合免疫组小鼠和rSj26 GST组小鼠肝脏中每只雌虫的虫卵数均显著高于对照组(P<0.01)。
与单独用pEGFP-Sj26或rSj26 GST免疫相比,pEGFP-Sj26与rSj26 GST联合免疫可增强对BALB/c小鼠的保护效果。