脂多糖与β葡聚糖对迟钝爱德华氏菌亚单位疫苗的免疫促进效果研究

doi:10.11924/j.issn.1000-6850.2013-1983

摘要/Abstract

摘要： 为了研究脂多糖与β葡聚糖的协同免疫促进效果，本论文重组表达了迟钝爱德华氏菌的一个高效亚单位疫苗候选蛋白甘油醛-3-磷酸脱氢酶（GAPDH），利用纯化的重组GAPDH（rGAPDH）蛋白单独或混合脂多糖和β葡聚糖免疫牙鲆，免疫后通过检测牙鲆抗体水平、免疫相关基因表达及免疫保护力三方面指标变化对脂多糖和酵母β葡聚糖的免疫促进效果进行研究。酶联免疫吸附试验（ELISA）检测结果显示，脂多糖和β葡聚糖与rGAPDH蛋白共免疫组牙鲆的抗体效价在免疫后第14和30天均显著高于单独免疫rGAPDH蛋白的牙鲆，在第30天时其效价为211.76；荧光定量PCR检测结果显示，相比PBS对照组，牙鲆在注射免疫后脾脏组织中IL-1β、IL-6、MHC-IIα、IgM、TCRα及C型溶菌酶6种免疫相关基因表达量均出现不同程度的上调，而其中脂多糖和β葡聚糖与rGAPDH蛋白共免疫组牙鲆在免疫后多个时间点其基因表达水平均显著高于单独免疫rGAPDH蛋白的牙鲆；免疫保护性实验结果同样显示，脂多糖和β葡聚糖与rGAPDH蛋白共免疫组牙鲆在攻毒后累计死亡率显著低于对照组和单独免疫rGAPDH蛋白组，直至第14天累计死亡率为30%，相对免疫保护率为68%。研究表明，脂多糖和β葡聚糖作为疫苗免疫刺激剂使用可显著增强牙鲆的免疫应答水平，提升亚单位疫苗的免疫效果。

关键词: 农户农药使用行为, 农户农药使用行为, 农药残留, 蔬菜出口产地, 食品安全

Abstract: In order to investigate the immunostimulatory effects of LPS and β-glucan, an highly effective vaccine candidate for Edwardsiella tarda known as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was recombinantly expressed in Escherichia coli, which was injected into Japanese flounder alone or containing LPS and β-glucan. After immunization, the antibody titer, expression of immune-related genes and immunological protection were detected. ELISA analysis of sera showed that the fish immunized with rGAPDH containing LPS and β-glucan had significant higher antibody titers than rGAPDH immunization group at day 14 and 30， which reached a value of 211.76 at day 30 after immunization. Simultaneously, the expression of 6 immune-related genes involved in innate and acquired responses, including IL-1β, IL-6, MHC-IIα, IgM, TCRα and C-type lysozyme were analyzed by quantitative PCR. The transcript levels of all selected genes displayed up-regulation after immunization, and immunization with rGAPDH containing LPS and β-glucan could induce significant higher levels than that induced only by rGAPDH. Moreover, to investigate the protection against infection, challenge was performed at day 30 after immunization. Fish immunized with rGAPDH containing LPS and β-glucan exhibited the highest relative percentage survival (RPS) of 67 % compared to the other groups. These results indicated that LPS and β-glucan employed as immunostimulatory molecules could significantly trigger the innate and acquired immune responses in Japanese flounder.

刁菁叶海斌于晓清许拉樊英李天保王淑君王娟. 脂多糖与β葡聚糖对迟钝爱德华氏菌亚单位疫苗的免疫促进效果研究[J]. 中国农学通报, 2014, 30(5): 46-52.

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