携带NDV HN基因疫苗的重组减毒鸡白痢沙门菌口服免疫原性研究

doi:10.11843/j.issn.0366-6964.2016.10.017

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (10): 2081-2088.doi: 10.11843/j.issn.0366-6964.2016.10.017

携带NDV HN基因疫苗的重组减毒鸡白痢沙门菌口服免疫原性研究

丁轲，余祖华，李蒙，郁川，尚珂，程相朝^*，陈桂华，廖成水，贾艳艳，汪洋，张春杰

（河南科技大学动物疫病与公共卫生重点实验室，洛阳 471003）

收稿日期:2016-03-22 出版日期:2016-10-23 发布日期:2016-10-23
通讯作者: 程相朝，E-mail: chengxch@126.com
作者简介:丁轲(1977-)，男，河南永城人，副教授，博士，主要从事动物传染病与动物微生态学研究，Tel:0379-64895698，E-mail: keding19@163.com
基金资助:
河南省重点科技攻关项目（152102110078）

Oral Immunogenicity of an Attenuated Salmonella pullorum Harbouring HN Gene Vaccine of Newcastle Disease Virus

DING Ke，YU Zu-hua，LI Meng，YU Chuan，SHANG Ke，CHENG Xiang-chao^*，CHEN Gui-hua，LIAO Cheng-shui, JIA Yan-yan, WANG Yang, ZHANG Chun-jie

（Key Laboratory of Animal Disease and Public Health，Henan University of Science and Technology，Luoyang 471003，China）

Received:2016-03-22 Online:2016-10-23 Published:2016-10-23

摘要/Abstract

摘要：

为研究携带新城疫病毒（NDV）血凝素-神经氨酸酶（HN）基因疫苗的重组减毒鸡白痢沙门菌的口服免疫原性，将携带NDV HN基因的重组质粒pcDNA3-HN电转化减毒鸡白痢沙门菌Δcrp C79-13，构建重组疫苗菌株Δcrp C79-13 (pcDNA3-HN)。将重组菌株Δcrp C79-13 (pcDNA3-HN)以1×10⁹CFU•只^-1口服免疫7日龄雏鸡，同时设PBS、Δcrp C79-13 (pcDNA3) 和NDV Ⅳ系疫苗免疫对照，于免疫后7、14、21、28、35 d 测定免疫雏鸡诱导的抗NDV 的HI抗体、鸡白痢沙门菌IgG抗体和肠道黏膜IgA抗体动态水平，同时测定免疫雏鸡的外周血淋巴细胞增殖水平并进行攻毒试验。结果表明，成功获得携带NDV HN基因疫苗的重组菌株Δcrp C79-13 (pcDNA3-HN)；在免疫后14~35 d，可诱导产生抗NDV的HI抗体，且在免疫后21 d时达到最高值；可诱导产生抗鸡白痢沙门菌的IgG抗体，且在免疫后28 d 达到最高值；Δcrp(C79-13 pcDNA3-HN)组肠道黏膜IgA抗体含量最高值略高于Δcrp C79-13 (pcDNA3)组，但差异不显著(P＞0.05)。在免疫28 d 以后，Δcrp(C79-13 pcDNA3-HN)组外周血淋巴细胞刺激指数极显著高于PBS对照组（P＜0.01)，显著高于ΔcrpC79-13(pCDNA3)组（P＜0.05)。用10³EID₅₀ NDV F₄₈E₉株攻毒，保护率达67%（10/15），用10⁸CFU鸡白痢沙门菌C79-13攻毒，保护率为100%。本研究结果表明重组减毒鸡白痢沙门菌Δcrp C79-13 (pcDNA3-HN)具有良好的口服免疫原性。

Abstract:

To study the oral immunogenicity of attenuated Salmonella pullorum，carried HN (haemagglutinin-neuraminidase) gene vaccine of Newcastle disease virus (NDV).In this study，the recombinant plasmid pcDNA3-HN was finally electro-transformed into attenuated Salmonella pullorum ΔcrpC79-13 to construct the recombinant strain ΔcrpC79-13 (pcDNA3-HN).Then the 7-day-old chickens were oral inoculated with 1×10⁹CFU recombinant strain ΔcrpC79-13 (pcDNA3-HN)，and the Δcrp C79-13 (pcDNA3)，PBS，NDV Ⅳvaccine were act as the control group.Then the dynamic level of HI antibody and intestinal mucosal IgA antibody against NDV were determined at 7，14，21，28，35 days post vaccination.At the same time，the level of peripheral blood lymphocyte proliferation of immune chicken and the challenge protection efficiency were determined.The results showed that the recombinant vaccine strains Δcrp C79-13 (pcDNA3-HN) carrying NDV HN gene vaccine was constructed successfully，which could induce HI antibody against NDV at 14-35 d after immunization and reach the highest value at 21 d after immunization.The content of intestinal mucosal IgA antibody in Δcrp (C79-13 pcDNA3-HN) group was slightly higher than that in Δcrp C79-13 (pcDNA3) group，but the difference was not significant (P> 0.05).The peripheral blood lymphocyte stimulation index of Δcrp (C79-13 pcDNA3-HN) group was significantly higher than that of the PBS control group (P<0.01)，and was slightly higher than that of the ΔcrpC79-13 (pCDNA3) group （P＜0.05) at 28 d after immunization.The immunity protection rate against strong virus was 67%（10/15） in experiment group with 10³EID₅₀NDV F₄₈E₉，the protection rate was 100% which challenged with 10⁸ CFU Salmonella pullorum C79-13 strain.The results showed that the recombinant attenuated Salmonella pullorum ΔcrpC79-13 (pcDNA3-HN) strains have good oral immunogenicity.

中图分类号:

S852.52

丁轲，余祖华，李蒙，郁川，尚珂，程相朝，陈桂华，廖成水，贾艳艳，汪洋，张春杰. 携带NDV HN基因疫苗的重组减毒鸡白痢沙门菌口服免疫原性研究[J]. 畜牧兽医学报, 2016, 47(10): 2081-2088.

DING Ke，YU Zu-hua，LI Meng，YU Chuan，SHANG Ke，CHENG Xiang-chao，CHEN Gui-hua，LIAO Cheng-shui, JIA Yan-yan, WANG Yang, ZHANG Chun-jie. Oral Immunogenicity of an Attenuated Salmonella pullorum Harbouring HN Gene Vaccine of Newcastle Disease Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(10): 2081-2088.

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携带NDV HN基因疫苗的重组减毒鸡白痢沙门菌口服免疫原性研究

Oral Immunogenicity of an Attenuated Salmonella pullorum Harbouring HN Gene Vaccine of Newcastle Disease Virus

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