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基于辛德毕斯病毒复制酶的 DNA 疫苗构建体,编码口蹄疫病毒特异性多价表位基因:在豚鼠中的免疫应答研究。

Sindbis virus replicase-based DNA vaccine construct encoding FMDV-specific multivalent epitope gene: studies on its immune responses in guinea pigs.

机构信息

FMD Research Centre, Indian Veterinary Research Institute, Bengaluru, Karnataka, India.

出版信息

Scand J Immunol. 2012 Oct;76(4):345-53. doi: 10.1111/j.1365-3083.2012.02733.x.

Abstract

Foot-and-mouth disease (FMD) is still a perennial global menace affecting livestock health and production. It is imperative to figure out new ways to curb this disease. In this study, a sindbis virus replicase-based DNA vaccine, pSinCMV-Vac-MEG990, encoding a multivalent epitope gene (representing tandemly linked VP1 C-terminal halves of three foot-and-mouth disease virus (FMDV) serotypes) was constructed. In vitro transfection studies in BHK-21 cells revealed that the construct was able to express FMDV-specific antigen but does not overproduce the antigen. Immunization of guinea pigs with the construct at dose rate of 10, 5, 2 and 1 μg per animal through intramuscular route showed significant neutralizing antibody induction at all doses against all serotype tested as compared to non-immunized controls. On viral challenge of guinea pigs 4 week post-immunization with 1000 GPID(50) of FMDV serotype A, it was observed that the immunization not only delayed the appearance and reduced the severity of FMD lesions significantly (P < 0.05) but also provided complete protection in several guinea pigs. In fact, two of six and one of six guinea pigs were completely protected in 10 and 5 μg immunized groups, respectively. These results suggest that the development of the replicase-based DNA vaccine may provide a promising approach as an alternative vaccine strategy for controlling FMD.

摘要

口蹄疫(FMD)仍然是一种全球性的常年威胁,影响着牲畜的健康和生产。必须找到新的方法来遏制这种疾病。在这项研究中,构建了一种基于辛德毕斯病毒复制酶的 DNA 疫苗 pSinCMV-Vac-MEG990,该疫苗编码一种多价表位基因(代表三种口蹄疫病毒(FMDV)血清型的 VP1 C 末端的串联连接的一半)。在 BHK-21 细胞中的体外转染研究表明,该构建体能够表达 FMDV 特异性抗原,但不会过度产生抗原。通过肌肉内途径以 10、5、2 和 1 μg/动物的剂量率用该构建体免疫豚鼠,与未免疫对照相比,所有测试的血清型均在所有剂量下均显示出显著的中和抗体诱导。在免疫接种后 4 周用 1000 GPID(50)的 FMDV 血清型 A 对豚鼠进行病毒攻毒,观察到免疫不仅显著延迟了 FMD 病变的出现并减轻了其严重程度(P < 0.05),而且在几只豚鼠中提供了完全保护。实际上,在 10 和 5 μg 免疫组中,有 2 只和 6 只豚鼠中的 1 只分别完全得到了保护。这些结果表明,基于复制酶的 DNA 疫苗的开发可能为控制 FMD 提供了一种有前途的替代疫苗策略。

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