Hall Roy A, Nisbet Debra J, Pham Kim B, Pyke Alyssa T, Smith Greg A, Khromykh Alexander A
Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, and Clinical Medical Virology Centre, University of Queensland, Brisbane 4072, Australia.
Proc Natl Acad Sci U S A. 2003 Sep 2;100(18):10460-4. doi: 10.1073/pnas.1834270100. Epub 2003 Aug 13.
A plasmid DNA directing transcription of the infectious full-length RNA genome of Kunjin (KUN) virus in vivo from a mammalian expression promoter was used to vaccinate mice intramuscularly. The KUN viral cDNA encoded in the plasmid contained the mutation in the NS1 protein (Pro-250 to Leu) previously shown to attenuate KUN virus in weanling mice. KUN virus was isolated from the blood of immunized mice 3-4 days after DNA inoculation, demonstrating that infectious RNA was being transcribed in vivo; however, no symptoms of virus-induced disease were observed. By 19 days postimmunization, neutralizing antibody was detected in the serum of immunized animals. On challenge with lethal doses of the virulent New York strain of West Nile (WN) or wild-type KUN virus intracerebrally or intraperitoneally, mice immunized with as little as 0.1-1 microg of KUN plasmid DNA were solidly protected against disease. This finding correlated with neutralization data in vitro showing that serum from KUN DNA-immunized mice neutralized KUN and WN viruses with similar efficiencies. The results demonstrate that delivery of an attenuated but replicating KUN virus via a plasmid DNA vector may provide an effective vaccination strategy against virulent strains of WN virus.
一种质粒DNA,其能在哺乳动物表达启动子的作用下,在体内指导库京(KUN)病毒全长感染性RNA基因组的转录,被用于对小鼠进行肌肉注射免疫。质粒中编码的KUN病毒cDNA在NS1蛋白(第250位脯氨酸突变为亮氨酸)上存在突变,先前已证明该突变可使KUN病毒在断奶小鼠中减毒。DNA接种后3 - 4天,从免疫小鼠的血液中分离出KUN病毒,这表明体内正在转录感染性RNA;然而,未观察到病毒诱导疾病的症状。免疫后19天,在免疫动物的血清中检测到中和抗体。在用致死剂量的西尼罗河(WN)病毒纽约强毒株或野生型KUN病毒进行脑内或腹腔内攻毒时,用低至0.1 - 1微克KUN质粒DNA免疫的小鼠能得到有效的疾病保护。这一发现与体外中和数据相关,表明KUN DNA免疫小鼠的血清能以相似效率中和KUN和WN病毒。结果表明,通过质粒DNA载体递送减毒但仍能复制的KUN病毒可能提供一种针对WN病毒强毒株的有效疫苗接种策略。