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表达S1和IL-2基因的双顺反子传染性支气管炎病毒DNA疫苗引发的强效免疫反应

[Potent immune responses elicited by a bicistronic IBV DNA vaccine expressing S1 and IL-2 gene].

作者信息

Tang Meng-Jun, Wang Hong-Ning, Zhou Sheng, Huang Yong, Liu Ping

机构信息

College of Animal Science & Technology, Sichuan Agricultural University, Yaan 625014, China).

出版信息

Wei Sheng Wu Xue Bao. 2007 Dec;47(6):1055-9.

Abstract

Abstract: Candidate IBV vaccines should elicit cellular responses as well as humoral responses. SI gene of avian infectious bronchitis virus and interleukin-2 (IL-2) gene from chicken were inserted into the bicistronic pIRES-EGFP/ DsRed plasmid. The pIRES-S1, pIRES-IL2 and pIRES-S1/IL-2 plasmid expressing or co-expressing S1 and IL-2 gene were constructed. Plasmids were transfected into the Vero cells by lipofectamine, and the expressed products were detected by RT-PCR and indirect immunofluorescence assay. The 7-day-old chickens were immunized intramuscularly with plasmids encapsulated by liposome and boosted three weeks later. Two weeks after boosting, chickens were challenged by virulent IBV strain. The results showed that coadministration of a plasmid expressing IL-2 with the S1 DNA vaccine led to only a marginal increase in humoral and T cell responses. However, immunization with the bicistronic plasmid pIRES-Sl/IL-2 that co-expressing S1 and IL-2 under control of a single promoter led to a dramatic augmentation of humoral and T cell responses. The protective efficacy could be significantly enhanced after injection with plasmids pIRES-S1/IL-2 or pIRES-S1 + pIRES-IL2. These results demonstrate that bicistronic DNA vaccine containing IL-2 elicit remarkably immune responses and suggest that optimal humoral and cellular responses priming requires the precise temporal and spatial codelivery of Ag and IL-2.

摘要

摘要

候选传染性支气管炎病毒(IBV)疫苗应能引发细胞免疫反应和体液免疫反应。将禽传染性支气管炎病毒的S1基因和鸡白细胞介素-2(IL-2)基因插入双顺反子pIRES-EGFP/DsRed质粒中。构建了表达或共表达S1和IL-2基因的pIRES-S1、pIRES-IL2和pIRES-S1/IL-2质粒。通过脂质体转染将质粒导入Vero细胞,并通过逆转录聚合酶链反应(RT-PCR)和间接免疫荧光试验检测表达产物。用脂质体包裹的质粒对7日龄雏鸡进行肌肉注射免疫,并在3周后进行加强免疫。加强免疫2周后,用强毒IBV毒株对雏鸡进行攻毒。结果表明,将表达IL-2的质粒与S1 DNA疫苗共同给药仅使体液免疫和T细胞反应略有增加。然而,用在单个启动子控制下共表达S1和IL-2的双顺反子质粒pIRES-S1/IL-2进行免疫导致体液免疫和T细胞反应显著增强。注射质粒pIRES-S1/IL-2或pIRES-S1 + pIRES-IL2后,保护效果可显著增强。这些结果表明,含IL-2的双顺反子DNA疫苗能引发显著的免疫反应,并表明最佳的体液免疫和细胞免疫反应启动需要抗原和IL-2精确的时间和空间共递送。

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