Bovine rotavirus nonstructural protein 4 produced by Lactococcus lactis is antigenic and immunogenic
Bovine rotavirus nonstructural protein 4 produced by Lactococcus lactis is antigenic and immunogenic
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DOI：
10.1128/aem.67.4.1423-1428.2001
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发表时间：
2001-04-01
影响因子：
4.4
通讯作者：
Corthier, G
中科院分区：
文献类型：
--
作者：
Enouf, V;Langella, P;Corthier, G
轮状病毒非结构蛋白4（NSP4）可引起小鼠腹泻。为了深入了解NSP4的生物学效应，需要大量生产这种蛋白质。我们首先尝试在大肠杆菌中生产这种蛋白质，但nsp4基因被证明是不稳定的。然后通过使用乳链菌肽控制的表达系统研究通常被认为是安全的生物体乳酸乳球菌在细胞内或细胞外产生NSP4的能力。在两个地点的乳酸乳球菌中均观察到重组NSP4（rNSP4）的产生。尽管分泌效率非常低，但用乳球菌信号肽和rNSP4之间的融合获得了最高水平的生产。将rNSP4分泌菌株的培养物注射到兔子中，并引发特异性免疫应答。在这些兔中产生的抗rNSP4抗体识别轮状病毒感染的MA104细胞中的NSP4。我们表明，乳酸乳球菌能够产生抗原性和免疫原性的rNSP4，因此是一个很好的生物体生产病毒抗原。
Rotavirus nonstructural protein 4 (NSP4) can induce diarrhea in mice. To get insight into the biological effects of NSP4, production of large quantities of this protein is necessary. We first tried to produce the protein in Escherichia coli, but the nsp4 gene proved to be unstable. The capacity of the generally regarded as safe organism Lactococcus lactis to produce NSP4 either intra- or extracellularly was then investigated by using the nisin-controlled expression system. Production of recombinant NSP4 (rNSP4) was observed in L, lactis for both locations. In spite of a very low secretion efficiency, the highest level of production was obtained with the fusion between a lactococcal signal peptide and rNSP4. Cultures of the rNSP4-secreting strain were injected into rabbits, and a specific immune response was elicited. The anti-rNSP4 antibodies produced in these rabbits recognized NSP4 in MA104 cells infected by rotavirus. We showed that L, lactis is able to produce antigenic and immunogenic rNSP4 and thus is a good organism for producing viral antigens.