Shuffling of pig interleukin-2 gene and its enhancing of immunity in mice to Pasteurella multocida vaccine.
Shuffling of pig interleukin-2 gene and its enhancing of immunity in mice to Pasteurella multocida vaccine.
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DOI：
10.1016/j.vaccine.2007.09.032
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发表时间：
2007-11
期刊：
影响因子：
5.5
通讯作者：
Zhao Xie;Hui Li;Jianlin Chen;Hua-Bing Zhang;Ying-Yu Wang;Qian Chen;Zhong-Zhong Zhao-Zhong;C. Cheng;Huan Zhang;Yi Yang;Hong-Ning Wang;R. Gao
中科院分区：
文献类型：
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作者：
Zhao Xie;Hui Li;Jianlin Chen;Hua-Bing Zhang;Ying-Yu Wang;Qian Chen;Zhong-Zhong Zhao-Zhong;C. Cheng;Huan Zhang;Yi Yang;Hong-Ning Wang;R. Gao
为了探索安全有效的促进动物抗感染免疫的佐剂，本实验将西藏猪IL-2 cDNA与人、牦牛、小鼠等IL-2 cDNA进行改组实验，研究改组后的IL-2（IL-2S）基因在体内对小鼠对多杀性巴氏杆菌免疫的影响。研究发现IL-2S蛋白比天然猪IL-2蛋白能显着促进猪淋巴母细胞的增殖。然后将IL-2S基因克隆到VR1020真核质粒(VRIL2S)中并用壳聚糖纳米粒子(CNP-VRIL2S)包裹。 21日龄雌性昆明小鼠分别肌肉接种CNP-VRIL2、CNP-VRIL2S和CNP-VR1020以及多杀性巴氏杆菌疫苗，并于接种后28天口服强毒多杀性巴氏杆菌进行攻击。每周采血，ELISA检测IgG、IgA、IgM、特异性抗体、IL-2、IL-4、IL-6的变化。接种并攻毒后，CNP-VRIL2S组的免疫球蛋白、特异性抗体和白细胞介素均较对照小鼠明显升高，10只免疫小鼠中有9只在攻毒后存活，而所有对照小鼠均出现严重症状和病变，最终死于感染。这些表明，包埋CNP的VRIL2S是一种新型安全有效的佐剂，可增强动物对传染性病原体的特异性免疫和抵抗力，这有助于开发极具前景的强力佐剂。
In order to explore the safe and effective adjuvant for promotion of immunity of animals against infection, the experiment was carried out to shuffle Tibet pig IL-2 cDNA with other IL-2 cDNA from human, yak and mouse, and the effect of shuffled IL-2 (IL-2S) gene in vivo was investigated on immunity of mice to Pasteurella multocida. The IL-2S protein was found to remarkably promote the proliferation of pig lymphoblasts than the native pig IL-2 protein. Then the IL-2S gene was cloned into VR1020 eukaryotic plasmid (VRIL2S) and enwrapped with chitosan nanoparticles (CNP-VRIL2S). Twenty-one day old female Kunming mice were muscularly inoculated respectively with the CNP-VRIL2, CNP-VRIL2S and CNP-VR1020 along with Pasteurella multocida vaccine, and orally challenged with virulent Pasteurella multocida on 28 days post-vaccination. The blood was weekly collected to detect the change of IgG, IgA, IgM, specific antibody, IL-2, IL-4 and IL-6 by ELISA. The immunoglobulins, specific antibody and interleukins significantly increased in CNP-VRIL2S group compared with the control mice after vaccination and challenge, and 9 of 10 immunized mice survived challenge, while the all control mice manifested severe symptoms and lesions, and finally died of infection. These indicated that VRIL2S entrapped with CNP is a novel safe and effective adjuvant to boost the specific immunity and resistance of animal against infectious pathogen, which could facilitate the development of highly promising powerful adjuvant.