Wang Xiaoquan, Jiang Ping, Deen Shovel, Wu Jiaqiang, Liu Xiaowen, Xu Jiarong
College of Veterinary Medicine, Nanjing Agriculture University, Nanjing 210095, China.
Avian Dis. 2003 Oct-Dec;47(4):1305-12. doi: 10.1637/6045.
The objective of the present study was to investigate the feasibility of a DNA vaccine and CpG oligodeoxynucleotide (ODN) to protect chickens against infectious bursal disease virus (IBDV) infection. Two plasmids DNA carrying VP2 genes of the very virulent (vv) strain of IBDV were constructed with reverse transcription-polymerase chain reaction and designated as pcDNA3.1-VP2 and pCI-VP2. The VP2 protein expressed in COS-7 cells transfected with the plasmid was confirmed by indirect immunofluorescence assay. Seven-day-old chickens were intramuscularly injected with the plasmids alone or plus commercial attenuated infectious bursal disease (IBD) vaccine or synthetic CpG ODN twice at weekly intervals. Chickens at 5 wk old were orally inoculated with vvIBDV strain 99J1 and observed for 7 days after challenge. Immunization with plasmid plus commercial attenuated IBD vaccine or CpG ODN conferred protection for 70%-80% of chickens, as evidenced by the absence of dinical signs, mortality, and atrophy in the cloacal bursa. About 25%-45% of chickens vaccinated with commercial attenuated IBD vaccine or pcDNA3.1-VP2 or pCI-VP2 plasmid alone had dinical signs and died after challenge. Furthermore, there were significantly different histopathologic lesion scores in the clocal bursae between the pcDNA3.1-VP2 or pCI-VP2 plus CpG or live vaccine and pcDNA3.1-VP2, pCI-VP2, or live vaccine vaccinated group. Enzyme-linked immunosorbent assay antibody titers in chickens vaccinated the constructs DNA plus live vaccine or CpG ODN were significantly higher than in those inoculated with the constructs or the live vaccine alone. These results suggest that coadministration of the constructed plasmid pcDNA3.1-VP2 or pCI-VP2 with CpG ODN or commercial attenuated IBD vaccine could protect chickens efficiently from direct vvIBDV challenge.
本研究的目的是探讨DNA疫苗和CpG寡脱氧核苷酸(ODN)保护鸡免受传染性法氏囊病病毒(IBDV)感染的可行性。用逆转录-聚合酶链反应构建了携带超强毒(vv)IBDV株VP2基因的两种质粒DNA,分别命名为pcDNA3.1-VP2和pCI-VP2。通过间接免疫荧光试验证实了用该质粒转染的COS-7细胞中表达的VP2蛋白。7日龄鸡单独肌肉注射质粒,或每周间隔两次肌肉注射质粒加商业减毒传染性法氏囊病(IBD)疫苗或合成的CpG ODN。5周龄鸡口服接种vvIBDV株99J1,攻毒后观察7天。质粒加商业减毒IBD疫苗或CpG ODN免疫可使70%-80%的鸡得到保护,表现为无临床症状、无死亡以及泄殖腔法氏囊无萎缩。单独接种商业减毒IBD疫苗或pcDNA3.1-VP2或pCI-VP2质粒的鸡中,约25%-45%在攻毒后出现临床症状并死亡。此外,pcDNA3.1-VP2或pCI-VP2加CpG或活疫苗组与pcDNA3.1-VP2、pCI-VP2或活疫苗接种组的泄殖腔法氏囊组织病理学病变评分有显著差异。接种构建体DNA加活疫苗或CpG ODN的鸡的酶联免疫吸附试验抗体效价显著高于单独接种构建体或活疫苗组。这些结果表明,构建的质粒pcDNA3.1-VP2或pCI-VP2与CpG ODN或商业减毒IBD疫苗联合使用可有效保护鸡免受vvIBDV的直接攻击。