Huang Jianmei, Chen Siying, Zhou Zhouyang, Sun Xiaoting, Haseeb Muhammad, Lakho Shakeel Ahmed, Zhang Yang, Liu Jiabin, Shah Muhammad Ali A, Song Xiaokai, Xu Lixin, Yan Ruofeng, Li Xiangrui
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R. China.
Departure of Parasitology and Microbiology, PMAS Arid Agriculture University, Rawalpindi 46000, Pakistan.
Poult Sci. 2021 May;100(5):101083. doi: 10.1016/j.psj.2021.101083. Epub 2021 Mar 2.
Eimeria tenella is a protozoan parasite endemic in chickens and is one of the causative agents of avian coccidiosis. The aim of this research was to determine if poly (D, L-lactide-co-glycolide) (PLGA) nanoparticles carrying recombinant TA4 protein of E. tenella (rEtTA4) could improve the level of protective immunity against E. tenella challenge. Recombinant TA4 protein was expressed and purified. Poly (D, L-lactide-co-glycolide) loaded with rEtTA4 (PLGA-rEtTA4) nanoparticles was prepared and was delivered to 2-week-old layer chickens via intramuscular inoculation. Chickens injected with PBS and PLGA nanoparticles were served as control groups. The rEtTA4 and PLGA-rEtTA4 nanoparticles induced changes of serum cytokines, IgY levels, and T lymphocytes subpopulation, and the protective efficacy against E. tenella challenge was evaluated. Results showed that both rEtTA4 and PLGA-rEtTA4 vaccination groups induced significantly higher levels of specific EtTA4 IgY antibody and IL-17 and higher proportion of CD8 T lymphocytes. However, no significant differences were observed in the proportion of CD4 T lymphocytes compared with the PBS control. Chickens immunized with rEtTA4 and PLGA-rEtTA4 prominently increased the BW gains and decreased oocyst output compared with chickens immunized with PBS and PLGA after oral challenge with E. tenella. Poly (D, L-lactide-co-glycolide) encapsulated rEtTA4 nanoparticles-immunized chickens significantly induced higher levels of interferon gamma, IL-6, and IL-17 and a little bit higher proportion of CD8 T lymphocytes compared with rEtTA4 subunit vaccine-immunized chickens. Thus, PLGA encapsulated rEtTA4 nanoparticles appeared to have great potential to enhance the immune response and improved the protective efficacy against E. tenella infection. Our results provided available protective subunit vaccine rEtTA4 and PLGA loaded with rEtTA4 nanoparticles against coccidiosis and suggested that PLGA nanoparticles could be an effective adjuvant to enhance the protective efficacy of rEtTA4 subunit vaccine.
柔嫩艾美耳球虫是鸡群中特有的一种原生动物寄生虫,也是禽球虫病的病原体之一。本研究的目的是确定携带柔嫩艾美耳球虫重组TA4蛋白(rEtTA4)的聚(D,L-丙交酯-共-乙交酯)(PLGA)纳米颗粒是否能提高对柔嫩艾美耳球虫攻击的保护性免疫水平。表达并纯化了重组TA4蛋白。制备了负载rEtTA4的聚(D,L-丙交酯-共-乙交酯)(PLGA-rEtTA4)纳米颗粒,并通过肌肉注射将其递送至2周龄的蛋鸡。注射PBS和PLGA纳米颗粒的鸡作为对照组。评估了rEtTA4和PLGA-rEtTA4纳米颗粒诱导的血清细胞因子、IgY水平和T淋巴细胞亚群的变化,以及对柔嫩艾美耳球虫攻击的保护效果。结果表明,rEtTA4和PLGA-rEtTA4疫苗接种组均诱导产生了显著更高水平的特异性EtTA4 IgY抗体和IL-17,以及更高比例的CD8 T淋巴细胞。然而,与PBS对照组相比,CD4 T淋巴细胞比例未观察到显著差异。与用PBS和PLGA免疫的鸡相比,用rEtTA4和PLGA-rEtTA4免疫的鸡在经柔嫩艾美耳球虫口服攻击后,体重增加显著,卵囊产量降低。与用rEtTA4亚单位疫苗免疫的鸡相比,用PLGA包裹的rEtTA4纳米颗粒免疫的鸡显著诱导产生了更高水平的干扰素γ、IL-6和IL-17,以及略高比例的CD8 T淋巴细胞。因此,PLGA包裹的rEtTA4纳米颗粒似乎具有增强免疫反应和提高对柔嫩艾美耳球虫感染保护效果的巨大潜力。我们的结果提供了针对球虫病的可用保护性亚单位疫苗rEtTA4和负载rEtTA4纳米颗粒的PLGA,并表明PLGA纳米颗粒可能是增强rEtTA4亚单位疫苗保护效果的有效佐剂。