Matos L, Muñoz M C, Molina J M, Rodríguez F, Perez D, Lopez A, Ferrer O, Hermosilla C, Taubert A, Ruiz A
Department of Animal Pathology, Faculty of Veterinary Medicine, University of Gran Canaria, Gran Canaria, Spain.
Deaprtment of Anatomy and Compared Anatomy Pathology, Faculty of Veterinary Medicine, University of Gran Canaria, Gran Canaria, Spain.
Vet Parasitol. 2017 Aug 15;242:1-9. doi: 10.1016/j.vetpar.2017.04.016. Epub 2017 Apr 15.
During the first schizogony, the goat coccidia Eimeria ninakohlyakimovae develops macroschizonts in lacteal duct endothelial cells, whose rupture leads to severe ileal damage and clinical signs during the prepatent period. The immune response elicited against early stages of the parasite development still requires to be investigated. In the present study we have evaluated immune reactions in goat kids primary- and challenged-infected with Eimeria ninakohlyakimovae, and sacrificed during prepatency (7days after challenge). The oocyst output during the primary infection, body weight and clinical condition of all the animals were examined and, at the end of the experiment, all the goat kids were euthanized and subjected to necropsy. Samples were taken from different sections of the ileum, colon and mesenteric lymph nodes (MLN) of primary- and challenged E. ninakohlyakimovae-infected animals. Intestinal leukocyte subpopulations were characterized in E. ninakohlyakimovae-infected mucosa and counts of lymphocytes, eosinophils, polymorphonuclear neutrophils (PMN), globular leukocytes and mast cells were recorded. Additionally, gene expression of caprine IL-2, IL-4, IL-10 and INFγ of ileal, colonic and MLN tissues were performed, as well as the immunohistochemical characterization of immune cells. The E. ninakohlyakimovae primary infection resulted in moderate to severe enteritis with different degrees of diarrhoea and was accompanied by high OPG counts and an increase of most immune cells analyzed when compared to uninfected control animals. Furthermore, eosinophil-, lymphocyte-, globular leukocyte- and mast cell-counts were significantly higher in the challenge group compared to the primary infected animals, whilst the opposite was true for PMN counts. The challenge infection was also associated with moderate increased levels of local mucosal IgA. Interestingly, the number of immature schizonts found at the ileal mucosa was statistically higher in the challenge infected group compared to the challenged control animals. Furthermore, in the challenged E. ninakohlyakimovae-infected animals a significantly higher number of mucosal CD4 and CD8 lymphocytes were observed, indicating that these T cell subpopulations might be involved in protective host immune response elicited against early stages of parasite development. The immune response was however very complex, as antigen presenting cells and other effector cell populations of the innate immune system, as well as certain cytokines, were involved. In summary, the results of this study contribute to the better understanding of local cellular and humoral immune responses against caprine E. ninakohlyakimovae, particularly during the prepatency.
在首次裂殖生殖过程中,山羊球虫尼氏艾美耳球虫在乳糜管内皮细胞中发育成大配子体,其破裂会导致严重的回肠损伤以及潜伏期的临床症状。针对寄生虫发育早期阶段引发的免疫反应仍有待研究。在本研究中,我们评估了初次感染和再次感染尼氏艾美耳球虫的山羊幼崽在潜伏期(攻毒后7天)的免疫反应,并对其实施安乐死。检查了所有动物初次感染期间的卵囊排出量、体重和临床状况,实验结束时,对所有山羊幼崽实施安乐死并进行尸检。从初次感染和再次感染尼氏艾美耳球虫的动物的回肠、结肠和肠系膜淋巴结(MLN)的不同部位采集样本。对感染尼氏艾美耳球虫黏膜中的肠道白细胞亚群进行了表征,并记录了淋巴细胞、嗜酸性粒细胞、多形核中性粒细胞(PMN)、球形白细胞和肥大细胞的数量。此外,还对回肠、结肠和MLN组织中山羊IL-2、IL-4、IL-10和INFγ的基因表达进行了检测,以及对免疫细胞进行了免疫组织化学表征。尼氏艾美耳球虫的初次感染导致中度至重度肠炎,并伴有不同程度的腹泻,与未感染的对照动物相比,其卵囊每克粪便排出量(OPG)较高,且大多数分析的免疫细胞数量增加。此外,与初次感染的动物相比,再次感染组的嗜酸性粒细胞、淋巴细胞、球形白细胞和肥大细胞数量显著更高,而PMN数量则相反。再次感染还与局部黏膜IgA水平适度升高有关。有趣的是,与再次感染的对照动物相比,再次感染组在回肠黏膜中发现的未成熟裂殖体数量在统计学上更高。此外,在再次感染尼氏艾美耳球虫的动物中,观察到黏膜CD4和CD8淋巴细胞数量显著更多,表明这些T细胞亚群可能参与了宿主针对寄生虫发育早期阶段引发的保护性免疫反应。然而,免疫反应非常复杂,因为抗原呈递细胞和先天免疫系统的其他效应细胞群体以及某些细胞因子都参与其中。总之,本研究结果有助于更好地理解针对山羊尼氏艾美耳球虫的局部细胞和体液免疫反应,特别是在潜伏期。