Division of Parasitology, Zoology Department, Faculty of Science, Beni-Suef University, Egypt.
Division of Immunity, Zoology Department, Faculty of Science, Beni-Suef University, Egypt.
Vet Parasitol. 2022 Sep;309:109772. doi: 10.1016/j.vetpar.2022.109772. Epub 2022 Jul 27.
Eimeriosis is a common parasitic disease in the chicken industry. The aim of this study was to assess the protective role of Hirudo extract antigens (HEA) against murine eimeriosis induced by Eimeria papillate. The oocyst output, developmental stages, goblet cells and oxidative stress, were investigated. Immunohistochemistry was used to detect anti-apoptotic Bcl marker and the number of both CD4 and CD25 cells in jejunal tissue, while ELISA was used to quantify TGF-β, IL-10 and IL-22 in jejunal tissue homogenate. Real-time PCR was also used to detect mRNA expression of mucin 2 (MUC2), inducible nitric oxide synthase (iNOS), IL-1β, IFN-γ, TNF-α, IL-6, and FoxP3. The most effective dose (5 µg/mice) reduced the oocyst output by 82.95 ± 1.02% (P ˂ 0.001). Similarly, the same dose reduced the jejunal developmental stages by 66.67 ± 0.49% (P ˂ 0.001). Furthermore, HEA therapy increased the number of jejunal goblet cells by 12.8 ± 1 (P ˂ 0.001) and the expression of MUC2 by 0.83 ± 0.06 (P ˂ 0.001). In contrast, TNF-α, IFN-γ, IL-6, iNOS, and IL-1β expression as well as apoptosis were reduced. The number of CD4 and CD25 in the jejunal tissue was increased (14.6 ± 1.2 (P ˂ 0.001), 6.84 ± 1 (P ˂ 0.01), respectively) after HEA therapy. The molecular analysis showed an increased expression of intestinal Foxp3 (3.2 ± 0.13 (P ˂ 0.001), while IL-22 was reduced (124 ± 10 (P ˂ 0.001)) versus an increase in TGF-β (250 ± 17 (P ˂ 0.01)) and IL-10 (236 ± 16 (P ˂ 0.001)) after HEA treatment in comparison to the non-treated infected group. With respect to the infected group, HEA reduced lipid peroxidation (LPO) (15.7 ± 1.12 (P ˂ 0.001)) and nitric oxide (NO) (13 ± 1.3 (P ˂ 0.001)) but increased reduced glutathione (GSH) (3.7 ± 0.26 (P ˂ 0.001)). In conclusion, HEA therapy protected against intestinal tissue damage by activation of CD4CD25Foxp3 cells which showed anti-inflammatory action. Hence, HEA can be recommended as a therapeutic treatment for eimeriosis.
球蛭素抗原（HEA）对由艾美耳球虫引起的鼠艾美耳球虫病的保护作用。研究卵囊产量、发育阶段、杯状细胞和氧化应激。免疫组织化学用于检测抗凋亡 Bcl 标记物和空肠组织中 CD4 和 CD25 细胞的数量，而 ELISA 用于定量空肠组织匀浆中的 TGF-β、IL-10 和 IL-22。实时 PCR 还用于检测粘蛋白 2 (MUC2)、诱导型一氧化氮合酶 (iNOS)、IL-1β、IFN-γ、TNF-α、IL-6 和 FoxP3 的 mRNA 表达。最有效剂量（5 µg/只小鼠）将卵囊产量降低 82.95% ± 1.02%（P ˂ 0.001）。同样，相同剂量将空肠发育阶段降低 66.67% ± 0.49%（P ˂ 0.001）。此外，HEA 治疗使空肠杯状细胞数量增加 12.8 ± 1（P ˂ 0.001），MUC2 表达增加 0.83 ± 0.06（P ˂ 0.001）。相反，TNF-α、IFN-γ、IL-6、iNOS 和 IL-1β 的表达以及细胞凋亡减少。空肠组织中 CD4 和 CD25 的数量增加（14.6 ± 1.2（P ˂ 0.001），6.84 ± 1（P ˂ 0.01））在 HEA 治疗后。分子分析显示，肠 Foxp3 的表达增加（3.2 ± 0.13（P ˂ 0.001），而 IL-22 减少（124 ± 10（P ˂ 0.001）），同时 TGF-β 增加（250 ± 17（P ˂ 0.01））和 IL-10（236 ± 16（P ˂ 0.001））与未治疗的感染组相比，在 HEA 治疗后。与感染组相比，HEA 降低了脂质过氧化（LPO）（15.7 ± 1.12（P ˂ 0.001））和一氧化氮（NO）（13 ± 1.3（P ˂ 0.001）），但增加了还原型谷胱甘肽（GSH）（3.7 ± 0.26（P ˂ 0.001））。总之，HEA 治疗通过激活具有抗炎作用的 CD4CD25Foxp3 细胞来保护肠道组织免受损伤。因此，HEA 可被推荐作为艾美耳球虫病的治疗方法。
