Kim Soo Joung, Sung Haan Woo, Han Jeong Hee, Jackwood Daral, Kwon Hyuk Moo
Department of Veterinary Medicine, Kangwon National University, Chunchon 200-701, Republic of Korea.
Vet Microbiol. 2004 Jun 10;101(1):39-51. doi: 10.1016/j.vetmic.2004.03.009.
Plasmid DNA vaccines pcDNA-VP2 expressing only VP2 protein and pcDNA-VP243 expressing VP2, VP4 and VP3 proteins of very virulent infectious bursal disease virus (vvIBDV) Korean SH/92 strain were constructed. The expression of viral proteins from constructed DNA vaccines was confirmed by an in vitro transcription/translation system and transfection in COS-7 cells. To investigate the protective efficacy of these DNA vaccines, 2-week-old chickens were injected intramuscularly and intraperitoneally with pcDNA-VP2 and pcDNA-VP243 twice at 2-week intervals. On week 2 after the second immunization, chickens were orally challenged with the vvIBDV SH/92 strain and observed for 10 days. Antibodies specific to IBDV were not detected by enzyme-linked immunosorbent assay in DNA vaccination groups before challenge but were induced after challenge. The immunized groups exhibited a higher survival rate and lower bursal atrophy as compared with the non-immunized groups after challenge. The survival rates of pcDNA-VP243 and pcDNA-VP2 groups were 70 and 50%, respectively, but the survival rate of challenge control group was only 10%. In the ConA-induced lymphocyte proliferation assay of peripheral blood and splenic lymphocytes, the immunized groups showed significantly higher proliferation responses (P< 0.05) than non-immunized groups. The maintenance of T cell proliferation activity in DNA vaccination groups may be closely related to the protection against vvIBDV. These results suggest that our plasmid DNA vaccines induced high protective immunity against vvIBDV, in which cell mediated immune response rather than humoral immune response seemed to contribute to the protection of chickens against vvIBDV infection.
构建了仅表达VP2蛋白的质粒DNA疫苗pcDNA-VP2和表达超强毒传染性法氏囊病病毒(vvIBDV)韩国SH/92株VP2、VP4和VP3蛋白的pcDNA-VP243。通过体外转录/翻译系统和在COS-7细胞中的转染,证实了构建的DNA疫苗中病毒蛋白的表达。为了研究这些DNA疫苗的保护效果,对2周龄的鸡每隔2周进行两次肌肉注射和腹腔注射pcDNA-VP2和pcDNA-VP243。在第二次免疫后第2周,用vvIBDV SH/92株对鸡进行口服攻毒,并观察10天。在攻毒前,DNA疫苗接种组通过酶联免疫吸附测定未检测到针对IBDV的抗体,但攻毒后诱导产生了抗体。攻毒后,与未免疫组相比,免疫组表现出更高的存活率和更低的法氏囊萎缩。pcDNA-VP243组和pcDNA-VP2组的存活率分别为70%和50%,但攻毒对照组的存活率仅为10%。在外周血和脾淋巴细胞的ConA诱导淋巴细胞增殖试验中,免疫组显示出比未免疫组显著更高的增殖反应(P<0.05)。DNA疫苗接种组T细胞增殖活性的维持可能与抗vvIBDV保护密切相关。这些结果表明,我们的质粒DNA疫苗诱导了针对vvIBDV的高保护性免疫,其中细胞介导的免疫反应而非体液免疫反应似乎有助于保护鸡免受vvIBDV感染。