摘要
目的研究日本血吸虫胰岛素受体-2(SJIR-2)纳米微球核酸疫苗对小鼠攻击感染的免疫保护效果。方法构建pEGFP-SJIR-2重组质粒,双酶切鉴定并测序,大量提取pEGFP-SJIR-2质粒,用壳聚糖(CHS)修饰的聚乳酸-羟基乙酸共聚物(PLGA)微球包裹,用包裹后的SJIR-2纳米微球免疫小鼠。将40只雌性BALB/c小鼠随机分为4组(n=10),分别注射PBS、空pEGFP质粒、CHS-PLGA微球和CHS-PLGA-pEGFP-SJIR-2微球各100μg免疫小鼠,末次免疫2周后,用日本血吸虫尾蚴攻击感染小鼠,每次免疫及感染尾蚴前收集各组小鼠血清,ELISA法检测各组小鼠血清内免疫球蛋白(IgG)水平的变化。小鼠感染尾蚴42 d后全部剖杀,收集成虫和虫卵并计算减虫率和减卵率。结果成功构建了pEGFP-SJIR-2重组质粒,与PBS组比较,CHS-PLGA-pEGFPSJIR-2组的成虫数和虫卵数差异有统计学意义(P<0.01)。CHS-PLGA-pEGFP-SJIR-2组的减虫率和减卵率分别为37.36%和46.82%,和PBS组相比,CHS-PLGA-pEGFP-SJIR-2组小鼠血清内IgG水平比明显增高(P<0.01),而pEGFP组和CHS-PLCA组成虫数和虫卵数与PBS组比较差异无统计学意义。结论 SJIR-2纳米微球核酸疫苗对感染血吸虫的BALB/c小鼠有一定的免疫保护效果,对其潜在的候选抗原疫苗的价值尚需深入研究。
Objective To research the immuno-protection of SJIR-2 DNA vaccine with nanometer microspheres a- gainst Schistosoma japonicum infection in mice. Methods To construct eukaryotic expression plasmid pEGFP- SJIR-2, identified by double digestion and sequenced delivery. The recombinant plasmid pEGFP-SJIR-2 was ex- tracted and was encapsulated into PLGA nanometer microspheres which were modified by CHS. 40 female BALB/c mice were randomly divided into 4 groups （n = 10） , each group of mice were injected with PBS, empty pEGFP plasmid, CHS-PLGA nanometer microspheres and CHS-PLGA-pEGFP-SJIR-2 nanometer microspheres 100 ixg, re- spectively. Two weeks after the last immunization, each mouse was infected by cercaria of Schistosoma japonicum, sera of mice in each group were collected before each immunization and challenge infection. ELISA was used to de- tect the change of IgG in each group of micesera. 42 days later, all mice were sacrificed. The adult worms and eggs were collected and counted, and the worm and egg reduction rates were calculated as well. Results The recombi- nant plasmid pEGFP-SJIR-2 was successfully constucted, and there was significant difference in the numbers of worm and egg between CHS-PLGA-pEGFP-SJIR-2 group and PBS group （ P 〈 0.01 ）. The worm andegg reduction rates in CHS-PLGA-pEGFP-SJIR-2 group were 37.36% and 46. 82% respectively. The IgG levels in mice sera of CHS-PLGA-pEGFP-SJIR-2 group were remarkably higher （P 〈0. 01 ） compared with PBS group. On the contrary, there was no significant difference between both pEGFP plasmid group and CHS-PLGA group in the numbers of worm and egg compared with PBS group. Conclusion SJIR-2 nanometer microspheres nucleic acid vaccine has some immuno-protection against Schistosoma japonicum infection in BALB/c mice, while it is worth further studying for it＇ s potential value to be a candidate antigen molecule of Schistosoma japonicum vaccine.
出处
《安徽医科大学学报》
CAS
北大核心
2016年第5期611-614,共4页
Acta Universitatis Medicinalis Anhui
基金
安徽高校省级自然科学研究项目(编号:KJ2013A183)
