Zhao Zhong-Zhong, Zhang Hua-Bing, Chen Qian, Su Dan, Xie Zhao, Wang Ying-Yu, Yang Yi, Wang Ze-Zhou, Li Jiang-Ling, Wu Kai-Yuan, Wang Hong-Ning, Meng Min-Jie, 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, Sichuan, PR China.
Comp Immunol Microbiol Infect Dis. 2009 May;32(3):191-205. doi: 10.1016/j.cimid.2007.10.001. Epub 2008 Feb 14.
A novel oligodeoxynuleotides containing 11 CpG motifs was synthesized and inserted into the VR1020 plasmid containing pig interleukin-6 (IL-6) gene (VPIL6) to construct recombinant plasmid, VPIL6C. The chitosan nanoparticles (CNP) were prepared by ionic cross linkage to entrap the VPIL6C (VPIL6C-CNP), VPIL6 (VPIL6-CNP) and CpG (CpG-CNP). 42-Day old female mice were divided into four groups and intramuscularly injected respectively with 6 pmol VPIL6C-CNP, VPIL6-CNP, CpG-CNP and VR1020-CNP along with the bivalent vaccines against the Pasteurellosis and hog cholera. The blood was weekly collected from mice after vaccination to detect the changes of immunoglobulins, specific antibodies, IL-2, IL-4, IL-6 and immune cells. 28 days after vaccination, the mice were orally challenged with virulent Pasteurella multocida. The results showed that in comparison with those of the control VR1020 group, the content of immunoglobulins, specific antibodies and interleukins significantly increased in the sera from the treated two groups (P<0.05). Meanwhile, the number of lymphocytes and monocytes also remarkably elevated in the treated groups (P<0.05). The immune responses of VPIL6C mice were notably stronger than those of VPIL6 and CpG group. The challenge results proved that the overall immunity was further promoted in the treated mice which resisted the challenge infection; while the control mice manifested evident symptoms and lesions, and died of infection. These suggested that VPIL6C-CNP could better promote the immunity and resistance of mice against Pasteurellosis than VPIL6-CNP and CpG-CNP, and facilitate the development of effective adjuvant to enhance the immunity of animal against infection.
合成了一种含有11个CpG基序的新型寡脱氧核苷酸，并将其插入含有猪白细胞介素-6（IL-6）基因的VR1020质粒（VPIL6）中构建重组质粒VPIL6C。通过离子交联法制备壳聚糖纳米粒（CNP）以包封VPIL6C（VPIL6C-CNP）、VPIL6（VPIL6-CNP）和CpG（CpG-CNP）。将42日龄雌性小鼠分为四组，分别肌肉注射6 pmol的VPIL6C-CNP、VPIL6-CNP、CpG-CNP和VR1020-CNP以及猪巴氏杆菌病和猪霍乱二价疫苗。接种疫苗后每周从小鼠采集血液，检测免疫球蛋白、特异性抗体、IL-2、IL-4、IL-6和免疫细胞的变化。接种疫苗28天后，用强毒多杀性巴氏杆菌对小鼠进行口服攻毒。结果显示，与对照VR1020组相比，处理的两组血清中免疫球蛋白、特异性抗体和白细胞介素的含量显著增加（P<0.05）。同时，处理组中淋巴细胞和单核细胞的数量也显著升高（P<0.05）。VPIL6C小鼠的免疫反应明显强于VPIL6和CpG组。攻毒结果证明，处理的小鼠抵抗攻毒感染的整体免疫力进一步提高；而对照小鼠表现出明显症状和病变，并死于感染。这些结果表明，与VPIL6-CNP和CpG-CNP相比，VPIL6C-CNP能更好地促进小鼠对巴氏杆菌病的免疫力和抵抗力，并有助于开发有效的佐剂以增强动物抗感染的免疫力。
