Animal Bioscience and Biotechnology Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, United States Department of Agriculture, Beltsville, MD, USA.
SEPPIC, 50 Boulevard National, Paris la Défense, 92257 La Garenne Colombes, France.
Poult Sci. 2022 Sep;101(9):102013. doi: 10.1016/j.psj.2022.102013. Epub 2022 Jun 17.
This study was conducted to develop a recombinant Eimeria elongation factor-1α (EF-1α)-vaccination strategy against Eimeria maxima (E. maxima) infection by co-administering with chicken IL-7 (chIL-7) or chicken NK-lysin peptide 2 (cNK-2) in commercial broiler chickens. Chickens were divided into the following 5 groups: control (CON, no Eimeria infection), nonimmunized control (NC, PBS plus Montanide ISA 78 VG), Vaccination 1 (VAC1, 100 µg of recombinant EF-1α plus Montanide ISA 78 VG), Vaccination 2 (VAC2, VAC1 plus 1 µg of chIL-7), and Vaccination 3 (VAC3, VAC2 plus 5 µg of cNK-2 peptide). The first immunization except the cNK-2 injection was performed intramuscularly on day 4, and the secondary immunization was given with the same concentration of components as the primary immunization 1 wk later. All chickens except the CON group were orally inoculated with freshly prepared E. maxima (1.0 × 10 oocysts per chicken) oocysts on Day 19. The results of the in vivo vaccination trial showed that chickens of all groups immunized with recombinant EF-1α antigen (VAC1, VAC2, and VAC3) showed higher serum antibody levels to EF-1α, and co-injection with chIL-7 further increased the serum IL-7 level in the VAC2 and VAC3 groups. Chickens in the VAC2 group showed significantly (P < 0.01) higher body weight gains at 6 and 9 d post-E. maxima challenge infection (dpi) with reduced gut lesions in the jejunum at 6 dpi. The VAC3 group showed reduced fecal oocyst shedding compared to the nonimmunized and infected chickens (NC). At 4 dpi, E. maxima infection significantly (P < 0.05) up-regulated the expression levels of proinflammatory cytokines (IL-β and IL-17F) and type Ι cytokines (IFN-γ and IL-10) in the jejunum (NC), but the expression of these cytokines was significantly (P < 0.05) down-regulated in the VAC1, VAC2, and VAC3 groups. Furthermore, E. maxima challenge infection significantly (P < 0.05) down-regulated the expressions of jejunal tight junction (TJ) proteins (Jam2 and Occludin) at 4 dpi, but their expression was up-regulated in the VAC2 and VAC3 groups. Collectively, these results show the protective effects of the EF-1α recombinant vaccine, which can be further enhanced by co-injection with chIL-7 or cNK-2 peptide against E. maxima infection.
本研究旨在通过在商业肉鸡中同时给予鸡白细胞介素-7(chIL-7)或鸡 NK 细胞溶素肽 2(cNK-2)肽,开发一种重组艾美耳球虫伸长因子-1α(EF-1α)疫苗接种策略来对抗堆型艾美耳球虫(E. maxima)感染。鸡被分为以下 5 组:对照组(CON,无艾美耳球虫感染)、非免疫对照组(NC,PBS 加 Montanide ISA 78 VG)、免疫 1 组(VAC1,100 µg 重组 EF-1α 加 Montanide ISA 78 VG)、免疫 2 组(VAC2,VAC1 加 1 µg chIL-7)和免疫 3 组(VAC3,VAC2 加 5 µg cNK-2 肽)。除 cNK-2 注射外,第一次免疫于第 4 天进行肌内注射,1 周后用与第一次免疫相同浓度的成分进行第二次免疫。除 CON 组外,所有鸡均于第 19 天经口接种新鲜制备的堆型艾美耳球虫(每鸡 1.0×10 个卵囊)卵囊。体内疫苗接种试验结果表明,所有用重组 EF-1α 抗原(VAC1、VAC2 和 VAC3)免疫的鸡均显示出更高的血清 EF-1α 抗体水平,并且与 chIL-7 共同注射进一步增加了 VAC2 和 VAC3 组的血清 IL-7 水平。与非免疫和感染鸡(NC)相比,VAC2 组在堆型艾美耳球虫感染后 6 和 9 天(dpi)体重增加显著(P < 0.01),6 dpi 时空肠病变减轻。与未免疫和感染鸡(NC)相比,VAC3 组粪便卵囊脱落减少。在 4 dpi 时,堆型艾美耳球虫感染显著(P < 0.05)上调了空肠中促炎细胞因子(IL-β 和 IL-17F)和 1 型细胞因子(IFN-γ 和 IL-10)的表达水平(NC),但在 VAC1、VAC2 和 VAC3 组中,这些细胞因子的表达显著(P < 0.05)下调。此外,在 4 dpi 时,堆型艾美耳球虫感染显著(P < 0.05)下调了空肠紧密连接(TJ)蛋白(Jam2 和 Occludin)的表达,但在 VAC2 和 VAC3 组中其表达上调。总之,这些结果表明 EF-1α 重组疫苗具有保护作用,与 chIL-7 或 cNK-2 肽共同注射可进一步增强其对堆型艾美耳球虫感染的保护作用。