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猪白细胞介素-2和白细胞介素-4对猪繁殖与呼吸综合征病毒（PRRSV）ORF5 DNA疫苗诱导的猪病毒特异性免疫反应的积极作用。

Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine.

作者信息

Tang Deyuan, Liu Jian, Li Chunyan, Zhang Hua, Ma Ping, Luo Xianfeng, Zeng Zhiyong, Hong Nining, Liu Xia, Wang Bin, Wang Feng, Gan Zhenlei, Hao Fei

机构信息

Department of Animal Science, Guizhou University, Guiyang 550025,

出版信息

J Vet Sci. 2014;15(1):99-109. doi: 10.4142/jvs.2014.15.1.99. Epub 2013 Oct 18.

DOI:10.4142/jvs.2014.15.1.99
PMID:24136204
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3973771/
Abstract

The purpose of this study was to investigate the effects of porcine interleukin (IL)-2 and IL-4 genes on enhancing the immunogenicity of a porcine reproductive and respiratory syndrome virus ORF5 DNA vaccine in piglets. Eukaryotic expression plasmids pcDNA-ORF5, pcDNA-IL-2, and pcDNA-IL-4 were constructed and then expressed in Marc-145 cells. The effects of these genes were detected using an indirect immunofluorescent assay and reverse transcription polymerase chain reaction (RT-PCR). Characteristic fluorescence was observed at different times after pcDNA- ORF5 was expressed in the Marc-145 cells, and PCR products corresponding to ORF5, IL-2, and IL-4 genes were detected at 48 h. Based on these data, healthy piglets were injected intramuscularly with different combinations of the purified plasmids: pcDNA-ORF5 alone, pcDNA-ORF5 + pcDNA-IL-2, pcDNA-ORF5 + pcDNA-IL-4, and pcDNA-ORF5 + pcDNA- IL-4 + pcDNA-IL-2. The ensuing humoral immune responses, percentages of CD4(+) and CD8(+) T lymphocytes, proliferation indices, and interferon-g expression were analyzed. Results revealed that the piglets co-immunized with pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2 plasmids developed significantly higher antibody titers and neutralizing antibody levels, had significantly increased levels of specific T lymphocyte proliferation, elevated percentages of CD4(+) and CD8(+) T lymphocytes, and significantly higher IFN-γ production than the other inoculated pigs (p < 0.05).

摘要

本研究旨在探讨猪白细胞介素（IL）-2和IL-4基因对增强猪繁殖与呼吸综合征病毒ORF5 DNA疫苗在仔猪体内免疫原性的影响。构建了真核表达质粒pcDNA-ORF5、pcDNA-IL-2和pcDNA-IL-4，然后在Marc-145细胞中进行表达。使用间接免疫荧光试验和逆转录聚合酶链反应（RT-PCR）检测这些基因的作用。在pcDNA-ORF5在Marc-145细胞中表达后的不同时间观察到特征性荧光，并在48小时检测到与ORF5、IL-2和IL-4基因对应的PCR产物。基于这些数据，给健康仔猪肌肉注射纯化质粒的不同组合：单独的pcDNA-ORF5、pcDNA-ORF5 + pcDNA-IL-2、pcDNA-ORF5 + pcDNA-IL-4以及pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2。分析随后的体液免疫反应、CD4(+)和CD8(+) T淋巴细胞百分比、增殖指数以及干扰素-γ表达。结果显示，与pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2质粒共同免疫的仔猪产生的抗体滴度和中和抗体水平显著更高，特异性T淋巴细胞增殖水平显著增加，CD4(+)和CD8(+) T淋巴细胞百分比升高，且干扰素-γ产生量显著高于其他接种仔猪（p < 0.05）。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/aff39634b902/jvs-15-99-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/d05ca64415a0/jvs-15-99-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/5ae3ec863109/jvs-15-99-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/fd2803365cca/jvs-15-99-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/5e120b91d53f/jvs-15-99-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/c3e119e493db/jvs-15-99-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/aff39634b902/jvs-15-99-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/d05ca64415a0/jvs-15-99-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/5ae3ec863109/jvs-15-99-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/fd2803365cca/jvs-15-99-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/5e120b91d53f/jvs-15-99-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/c3e119e493db/jvs-15-99-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b217/3973771/aff39634b902/jvs-15-99-g006.jpg

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