鸡白细胞介素-18真核表达载体构建与ILTV弱毒疫苗联合免疫

为了探索鸡IL-18在传染性喉气管炎病毒(ILTV)弱毒疫苗中的免疫佐剂作用,将鸡IL-18(ChIL-18)克隆至真核表达载体pcDNA3.1(+)中,构建重组真核表达质粒pcDNA3.1/ChIL-18(简称pIL-18)。用pIL-18、ILTV弱毒疫苗及其两者联合分别免疫21日龄SPF雏鸡,免疫后每周抽取外周血及外周抗凝血,应用ELISA和流式细胞仪分别检测免疫鸡的体液免疫及细胞免疫水平。免疫后第4周用100 ELD50ILTV强毒进行攻击。结果pIL-18与ILTV弱毒疫苗联合免疫组的淋巴细胞亚群CD3+、CD3+CD4+、CD3+CD8+明显高于ILTV弱毒疫苗免疫组(P<0.01),ILTV强毒攻毒后发病鸡数量少,并且发病症状较轻。

Construction of Eukaryotic Expression Vector of Chicken Interleukin IL-18 and Its Combined Immunity with ILTV Attenuated Vaccine

To study the adjuvant effect of chicken interleukin-18 (ChIL-18) on Infectious Laryngotracheitis Virus (ILTV) attenuated vaccine, chicken IL-18 gene was cloned and transformed into eukaryotic expressing vector pcDNA3.1 (+) to generate an expressing plasmid pcDNA3.1/ChIL-18 (pIL-18). The 21-day-old SPF chickens were immunized with pIL-18, ILTV attenuated vaccine and combined immunity with both of them respectively. After that, peripheral blood and peripheral anti-coagulation were detected by flow cytometry and ELISA method at humeral immunity and cellular immunity level each week, respectively. Virulence 100 ELD_(50) ILTV was given to the chickens at 4 weeks after vaccination. The results showed that chickens vaccinated with combined immunity of pIL-18 and ILTV attenuated vaccine had an significancy increase in the percentage of CD3~+, CD~3+CD4~+, and CD3~+CD8~+ subgroups of peripheral blood compared with that only immunized by ILTV attenuated vaccine. Chickens were given virulence to.have fewer diseases and milder symptom than others.