H5亚型高致病性禽流感病毒HA基因重组新城疫病毒活载体疫苗接种后气管黏膜损伤的研究

Thacheal epithelium lesion of chickens vaccinated with recombinant NDV La Sota vaccine strain expressing HA gene of H5 highly pathogenicity Avian influenza virus

为了研究H5亚型高致病性禽流感病毒HA基因重组新城疫病毒活载体疫苗[rLa Sota-HA(GD)]的免疫机理,用rLa Sota-HA(GD)免疫2日龄SPF雏鸡,分别在接种后的1,3,5,10,15,18天取气管进行电镜、光镜观察,利用免疫组化染色(IHC)方法检测病毒在气管内的分布。结果表明:接种rLa Sota-HA(GD)后第1天纤毛细胞有脱纤毛现象,杯状细胞破碎,黏液成分增多,黏膜下层的纤维裸露在表面上,并可看到纤毛细胞的纤毛上有球形粒子,杯状细胞、基细胞大量增生;接种后第18天纤毛细胞脱纤毛区域进一步增大。接种后第3天黏膜下层水肿,淋巴细胞、粒细胞、浆细胞浸润,黏膜上皮形成空泡状,表层由数层不成熟的细胞组成;接种后第10天腺体腔闭塞。免疫组化染色可见气管固有层细胞呈阳性反应,棕色颗粒出现在胞浆中。结果说明气管黏膜的损伤是机体重要的防御机制和免疫机制的表现。

Vaccination was given by eyedrop and intranasally instillation with rLa Sota - HA （ GD ） （ 2 × 10^6 EID50/chicken ） to two - day - old SPF chicken after 1 , 3 , 5 , 10 , 15 , 18 days. The tracbeas were collected. Uhrastructural changes in the tracheal epithelium were examined by scanning electron microscopy. On day 1 postvaccation （PV）scanning electron microscopy revealed hypertrophy of goblet cell and small patches of the deciliated epithelium around the disoriented and deformed cilia. On days 1 PV hyperplasia of goblet cells accompanied by an increase in mucus. On days 1 PV globular particles found in areas of the trachea epithelium with tips of cilia adhering to them,goblel cells and basal cell regenerated. Non - ciliated plaques were observed until day 18 PV. But they gradually decreased in size. Light microscopy showed small vacuoles containing lymphocytes and heterophils plasma cells in the epithelial layer. The subrsueosa is slightly cdemalous, immature epithelium proliferated in some areas. The trachea is lined with several layers of immuture cells on days 3 PV. There is hypcrplasia of mucous gland cells eausing narrowong or obliteration of glandular lumina in immunohistochemical（IHC） staining on days 10 PV. Speeilfic staining was localized in the submucosa cells. This response is probably an attempt by the mucoeillary defense mechanism.