小鹅瘟病毒VP3真核表达质粒与弱毒疫苗诱导鹅体免疫应答的比较

中国农业科学 ›› 2011, Vol. 44 ›› Issue (3): 595-603 .

小鹅瘟病毒VP3真核表达质粒与弱毒疫苗诱导鹅体免疫应答的比较

朱德康,黎敏,车茜,程安春,汪铭书,陈孝跃

（四川农业大学动物医学院禽病防治研究中心）

收稿日期:2009-01-14 修回日期:2010-10-22 出版日期:2011-02-01 发布日期:2011-02-01
通讯作者: 程安春,汪铭书

Comparative Study on Immune Responses of Goose Induced by pcDNA-GPV-VP3 and Attenuated Vaccine

ZHU De-kang, LI Min, CHE Qian, CHENG An-chun, WANG Ming-shu, CHEN Xiao-yue

（四川农业大学动物医学院禽病防治研究中心）

Received:2009-01-14 Revised:2010-10-22 Online:2011-02-01 Published:2011-02-01
Contact: CHENG An-chun, WANG Ming-shu

摘要/Abstract

摘要：

【目的】比较小鹅瘟病毒（GPV）VP3基因疫苗（pcDNA-GPV-VP3）和弱毒疫苗免疫鹅体内的免疫应答,以期为进一步研究和阐明pcDNA-GPV-VP3免疫发生机理、免疫持续期等提供基础数据。【方法】将pcDNA-GPV-VP3和小鹅瘟弱毒疫苗分别免疫30日龄四川白鹅,用免疫组织化学方法检测免疫鹅体内GPV抗原的分布,用淋巴细胞增殖实验和间接ELISA分别检测免疫鹅的细胞免疫水平和血清抗体滴度,比较GPV基因疫苗与弱毒疫苗诱导鹅体免疫应答的能力。【结果】①弱毒疫苗组于免疫鹅的心、肝、脾、肺、肾、法氏囊、胸腺、哈氏腺、十二指肠、空肠、回肠、直肠、盲肠、胰腺、大脑及注射部位肌肉均中检测到GPV抗原;基因疫苗组于免疫鹅的心、十二指肠、空肠、回肠、直肠、盲肠及注射部位肌肉中检测到GPV抗原。②弱毒疫苗免疫雏鹅外周血T淋巴细胞14 d后对ConA的反应开始增强,35 d达到最高值后下降;基因疫苗免疫雏鹅外周血T淋巴细胞OD值先下降,14 d后开始上升并高于空白质粒对照鹅和PBS对照鹅,35 d时达最大值,以后又逐渐下降,第21-63天极显著（P＜0.01）高于PBS对照鹅和空白质粒对照鹅,第21-63 天时极显著（P＜0.01）高于弱毒疫苗免疫鹅。③弱毒疫苗免疫鹅血清抗体第3天开始高于PBS对照鹅,第28天达到最大值后逐渐下降;基因疫苗免疫鹅血清体从第14天开始高于PBS对照组,第28天达最大值,第21-217天显著（P＜0.05）或极显著（P＜0.01）高于PBS和空白质粒对照鹅;基因疫苗免疫鹅血清抗体除第28天极显著(P＜0.01)高于GPV弱毒疫苗免疫鹅外,其余时间与弱毒疫苗免疫鹅差异不显著。【结论】pcDNA-GPV-VP3免疫雏鹅后能在免疫部位皮肤、心肌和各肠段中表达并能够诱导鹅体产生良好的细胞免疫和体液免疫,基因疫苗诱导鹅体产生免疫应答的能力优于弱毒疫苗,结果为阐述pcDNA-GPV-VP3的免疫机制和临床应用提供了数据资料。

关键词: 小鹅瘟病毒, 基因疫苗, 弱毒疫苗, 鹅, 细胞免疫, 抗体

Abstract:

【Objective】 The immune response of immunized ducklings induced by goose parvovirus VP3 gene vaccine（pcDNA-GPV-VP3）and attenuated vaccine was compared for providing experimental data for explaination of immune mechanisms and immune duration of pcDNA-GPV-VP3.【Method】 The 30-day-old Sichuan White Geese were immunized with pcDNA-GPV-VP3 gene vaccine and attenuated vaccine of GPV. The antigens of GPV were detected by immunohistochemistry method. To compare the immune response ability of goose induced by gene vaccine and attenuated vaccine, the cell immunity effect and serum antibody titer were detected by lymphocyte proliferation assay and indirect ELISA, respectively. 【Result】 The GPV antigen was detected in the heart, liver, spleen, lung, kidney, bursa of Fabricius, thymus, Harder's glands, duodenum, jejunum, ileum, rectum, appendix, pancreas, cerebrum and injection muscle of goose in attenuated vaccine group. In gene vaccine group, the GPV antigen was detected in the heart, duodenum, jejunum, ileum, rectum, appendix, pancreas, cerebrum and injection muscle of goose. The action of T lymphocyte to ConA was enhanced in peripheral blood of immunized goose with attenuated vaccine from 14th day. It reached the highest at 35th day. The OD value of T lymphocyte descended at first in the gene vaccine group. It rose at 14th day and reached the highest at 35th day. The value in 21-63 d was higher than that of the control group and attenunated vaccine group, the difference was extremely significant (P＜0.01). The serum antibody titer of goose immunized with attenuated vaccine was higher than that of PBS control group from 3rd day. It reached the highest at 28th day. The serum antibody titer of goose immunized with gene vaccine was higher than that of PBS control group from 14th day. It reached the highest at 28th day. The value in 21-217 d was higher than that of the control group, the difference was significant (P＜0.05) or extremely significant (P＜0.01). The serum antibody titer of goose immunized with gene vaccine and attenuated vaccine had no obvious difference at any time except that in 28th day, and the difference was extremely significant (P＜0.01). 【Conclusion】The pcDNA-GPV-VP3 gene vaccine could express in the immunized muscle, heart and intestines of goose and induced favorable cell immunity and humoral immunity. The immune response ability induced by gene vaccine has more advantages than by attenuated vaccine. It is helpful to explain the immunologic mechanism and clinical application of pcDNA-GPV-VP3.

Key words: gosling plague virus, gene vaccine, attenuated vaccine, geese, cell immunity, antibody

朱德康,黎敏,车茜,程安春,汪铭书,陈孝跃 . 小鹅瘟病毒VP3真核表达质粒与弱毒疫苗诱导鹅体免疫应答的比较[J]. 中国农业科学, 2011, 44(3): 595-603 .

ZHU De-kang,LI Min,CHE Qian,CHENG An-chun,WANG Ming-shu,CHEN Xiao-yue
. Comparative Study on Immune Responses of Goose Induced by pcDNA-GPV-VP3 and Attenuated Vaccine
[J]. Scientia Agricultura Sinica, 2011, 44(3): 595-603 .

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