RABIES VIRUS NUCLEOPROTEIN EXPRESSED IN AND PURIFIED FROM INSECT CELLS IS EFFICACIOUS AS A VACCINE
RABIES VIRUS NUCLEOPROTEIN EXPRESSED IN AND PURIFIED FROM INSECT CELLS IS EFFICACIOUS AS A VACCINE
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DOI：
10.1073/pnas.88.5.2001
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发表时间：
1991-03-01
影响因子：
11.1
通讯作者：
KOPROWSKI, H
中科院分区：
文献类型：
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作者：
FU, ZF;DIETZSCHOLD, B;KOPROWSKI, H
将狂犬病毒Evelyn-Rokitnicki-Abelseth株核蛋白N的RNA基因cDNA拷贝克隆到杆状病毒中。重组杆状病毒在夜蛾细胞中大量表达N蛋白。利用亲和层析法从感染的褐夜蛾细胞中提取N蛋白，得到接近均匀的N蛋白。纯化的N蛋白与32种识别天然狂犬病毒核糖核蛋白的单克隆抗体中的31种反应。与核糖核蛋白一样，纯化的N蛋白是一种能够诱导病毒特异性辅助性T细胞的主要抗原。在灭活Evelyn-Rokitnicki-Abelseth病毒疫苗加强接种之前，用纯化的N蛋白引发小鼠，导致病毒中和抗体的产生增加20倍。用纯化的N蛋白免疫小鼠后，小鼠产生强烈的抗核糖核蛋白抗体反应，保护小鼠免受狂犬病病毒的致命攻击。这些数据表明，在昆虫细胞中表达的N蛋白在抗原性和免疫原性上与真正的狂犬病毒核糖核蛋白相当，因此代表了人类和动物大规模免疫狂犬病的有效和经济疫苗的潜在来源。
A cDNA copy of the RNA gene that encodes the nucleoprotein N of rabies virus Evelyn-Rokitnicki-Abelseth strain was cloned into baculovirus. The recombinant baculovirus expressed the N protein abundantly in Spodoptera frugiperda cells. The N protein was extracted from infected Spodoptera frugiperda cells and purified to near homogeneity by affinity chromatography. The purified N protein reacted with 31 of 32 monoclonal antibodies that recognize native rabies virus ribonucleoprotein. Like the ribonucleoprotein, the purified N protein was a major antigen capable of inducing virus-specific helper T cells. Priming of mice with the purified N protein prior to a booster inoculation with inactivated Evelyn-Rokitnicki-Abelseth virus vaccine resulted in a 20-fold increase in the production of virus-neutralizing antibodies. After immunization with the purified N protein, mice developed a strong anti-ribonucleoprotein antibody response and were protected against a lethal challenge of rabies virus. These data indicate that the N protein expressed in insect cells is antigenically and immunogenically comparable to the authentic rabies virus ribonucleoprotein and therefore represents a potential source of an effective and economical vaccine for large-scale immunization of humans and animals against rabies.