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共表达堆型艾美耳球虫3-1E基因和成熟的CHIl-15基因的DNA疫苗诱导细胞免疫应答

Induction of Cellular Immune Response by DNA Vaccine Coexpressing E. acervulina 3-1E Gene and Mature CHIl-15 Gene.

作者信息

Ma Dexing, Ma Chunli, Gao Mingyang, Li Guangxing, Niu Ze, Huang Xiaodan

机构信息

College of Veterinary Medicine, Northeast Agricultural University, No. 59 Mucai Street, Gongbin Road, Xiangfang District, Harbin 150030, China.

出版信息

J Parasitol Res. 2012;2012:654279. doi: 10.1155/2012/654279. Epub 2012 Jun 17.

Abstract

We previously reported that the chimeric DNA vaccine pcDNA-3-1E-linker-mChIL-15, fused through linking Eimeria acervulina 3-1E encoding gene and mature chicken IL-15 (mChIL-15) gene with four flexible amino acid SPGS, could significantly offer protection against homologous challenge. In the present study, the induction of cellular immune response induced by the chimeric DNA vaccine pcDNA-3-1E-linker-mChIL-15 was investigated. Spleen lymphocyte subpopulations were characterized by flow cytometric analysis. The spleen lymphocyte proliferation assays were measured by 3-[4,5-dimethylthiazol-2-y1]-2,5-diphenyltetrazolium bromide (MTT) method. The mRNA profiles of ChIL-2 and ChIFN-γ in spleen were characterized by means of real-time PCR. Chickens immunized with pcDNA-3-1E-linker-mChIL-15 exhibited significant upregulated level of ChIL-2 and ChIFN-γ transcripts in spleen following two immunizations compared with chickens in other groups (P < 0.01). In comparison with pcDNA3.1-immunized and control groups, lymphocyte proliferation, percentage of CD8α(+) cell, and levels of ChIL-2 and ChIFN-γ transcripts in the group immunized with pcDNA-3-1E-linker-mChIL-15 were significantly increased on day 6 following challenge (P < 0.05, P < 0.01, and P < 0.01, resp.). Our data suggested that the fusion antigen 3-1E-linker-mChIL-15 could be a potential candidate for E. acervulina vaccine development.

摘要

我们之前报道过,嵌合DNA疫苗pcDNA-3-1E-linker-mChIL-15通过将堆型艾美耳球虫3-1E编码基因与成熟鸡白细胞介素-15(mChIL-15)基因经四个柔性氨基酸SPGS连接融合而成,能显著提供针对同源攻击的保护。在本研究中,对嵌合DNA疫苗pcDNA-3-1E-linker-mChIL-15诱导的细胞免疫应答进行了研究。通过流式细胞术分析对脾淋巴细胞亚群进行表征。用3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐(MTT)法测定脾淋巴细胞增殖试验。通过实时PCR对脾脏中ChIL-2和ChIFN-γ的mRNA谱进行表征。与其他组的鸡相比,用pcDNA-3-1E-linker-mChIL-15免疫的鸡在两次免疫后脾脏中ChIL-2和ChIFN-γ转录本水平显著上调(P<0.01)。与pcDNA3.1免疫组和对照组相比,在攻击后第6天,用pcDNA-3-1E-linker-mChIL-15免疫的组中淋巴细胞增殖、CD8α(+)细胞百分比以及ChIL-2和ChIFN-γ转录本水平显著增加(分别为P<0.05、P<0.01和P<0.01)。我们的数据表明,融合抗原3-1E-linker-mChIL-15可能是堆型艾美耳球虫疫苗开发的潜在候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/99d1/3382946/8a6d37676c81/JPR2012-654279.001.jpg

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