Zhang Huan, Cheng Chi, Zheng Min, Chen Jian-Lin, Meng Ming Jie, Zhao Zhong-Zhong, Chen Qian, Xie Zhao, Li Jiang Ling, Yang Yi, Shen Yi, Wang Hong-Ning, Wang Ze-Zhou, Gao Rong
Key Laboratory for Bio-Resource and Eco-Environment of Ministry Education, Bioengineering Research Center for Animal Disease Prevention and Control, Life Science College, Sichuan University, Chengdu 610064, Sichuan, PR China.
Vaccine. 2007 Oct 10;25(41):7094-101. doi: 10.1016/j.vaccine.2007.07.050. Epub 2007 Aug 15.
Experiments were conducted to investigate the effect of a fusion gene of porcine IL-4 and IL-6 (PIL4/IL6) packaged with chitosan nanoparticles (CNPs) in terms of the development of a novel effective adjuvant. The IL4/PIL6 fusion gene was constructed and inserted into a eukaryotic expression vector. The plasmid was bound to CNP and then utilized to orally inoculate 21-day-old female Kunming mice that simultaneously received intramuscular injection of inactivated Escherichia coli vaccine. At 35 days post-vaccination, the mice were challenged by oral feeding with virulent O139: K88 strain EPEC E. coli bacteria. Compared with those of control mice, the content of immunoglobulins and specific antibodies to E. coli increased significantly in the sera of mice immunized with VPIL4/IL6-CNP (P<0.05). Furthermore, the levels of IL-2, IL-4 and IL-6 increased remarkably in the sera of immunized mice (P<0.05). After challenge, these immunological markers were elevated to different degrees in the mice immunized with the fusion gene construct (IL4/VPIL6-CNP) or individual plasmids (VPIL4+VPIL6-CNP). The immunized mice all survived the challenge and did not show any symptoms or lesion, whereas the VR1020-CNP control mice manifested obvious clinical symptoms and hemorrhagic lesions in the digestive tracts. These results demonstrated that VPIL4/IL6 entrapped with CNP is a novel promising adjuvant to promote specific immunity and resistance of animals against infectious pathogen.
开展实验以研究用壳聚糖纳米颗粒（CNPs）包裹的猪白细胞介素4和白细胞介素6融合基因（PIL4/IL6）作为新型有效佐剂的效果。构建了IL4/PIL6融合基因并将其插入真核表达载体。将该质粒与CNP结合，然后用于口服接种21日龄雌性昆明小鼠，同时这些小鼠接受肌肉注射灭活大肠杆菌疫苗。在接种疫苗后35天，通过口服强毒O139:K88株肠致病性大肠杆菌对小鼠进行攻毒。与对照小鼠相比，用VPIL4/IL6-CNP免疫的小鼠血清中免疫球蛋白和大肠杆菌特异性抗体的含量显著增加（P<0.05）。此外，免疫小鼠血清中白细胞介素2、白细胞介素4和白细胞介素6的水平显著升高（P<0.05）。攻毒后，在用融合基因构建体（IL4/VPIL6-CNP）或单个质粒（VPIL4+VPIL6-CNP）免疫的小鼠中，这些免疫标志物均有不同程度的升高。免疫小鼠在攻毒后全部存活，未出现任何症状或病变，而VR1020-CNP对照小鼠表现出明显的临床症状和消化道出血性病变。这些结果表明，用CNP包裹的VPIL4/IL6是一种新型且有前景的佐剂，可促进动物针对传染性病原体的特异性免疫和抵抗力。
