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Visualisation and characterisation of mononuclear phagocytes in the chicken respiratory tract using CSF1R -transgenic chickens

机译:使用CSF1R转基因鸡对鸡呼吸道中单核吞噬细胞的可视化和鉴定

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The respiratory tract is a key organ for many avian pathogens as well as a major route for vaccination in the poultry industry. To improve immune responses after vaccination of chickens through increased uptake of vaccines and targeting to antigen presenting cells, a better understanding of the avian respiratory immune system is required. Transgenic MacReporter birds were used expressing a reporter gene (eGFP or mApple) under the control of the CSF1R promoter and enhancer in cells of the mononuclear phagocyte (MNP) lineage to visualize the ontogeny of the lymphoid tissue, macrophages and dendritic cells, in the trachea, lung and air sac of birds from embryonic day 18–63?weeks of age. Small aggregates of CSF1R-transgene+ cells start to form at the openings of the secondary bronchi at 1?week of age, indicative of the early development of the organised bronchus-associated lymphoid tissue. Immunohistochemical staining revealed subpopulations of MNPs in the lung, based on expression of CSF1R-transgene, CD11, TIM4, LAMP1, and MHC II. Specialised epithelial cells or M cells covering the bronchus-associated lymphoid tissue expressed CSF1R-transgene and type II pneumocytes expressed LAMP1 suggesting that these epithelial cells are phagocytic and transcytose antigen. Highly organised lymphoid tissue was seen in trachea from 4?weeks onwards. Throughout the air sacs at all ages, CSF1R-transgene+ cells were scattered and at later stages, CSF1R-transgene+ cells lined capillaries. These results will serve as a base for further functional characterization of macrophages and dendritic cells and their role in respiratory diseases and vaccine responses.
机译:呼吸道是许多禽类病原体的关键器官,也是家禽业进行疫苗接种的主要途径。为了通过增加对疫苗的摄取和靶向抗原呈递细胞来改善鸡接种疫苗后的免疫反应,需要对鸟类呼吸免疫系统有更好的了解。使用转基因MacReporter鸟在单核吞噬细胞(MNP)谱系的细胞中在CSF1R启动子和增强子的控制下表达报告基因(eGFP或mApple),以观察气管中淋巴样组织,巨噬细胞和树突状细胞的存在。胚胎18-63周龄的鸟类的肺,气囊和气囊。 CSF1R-transgene +细胞的小聚集体在1周龄时在次级支气管的开口处开始形成,表明有组织的与支气管相关的淋巴样组织的早期发育。免疫组织化学染色显示,根据CSF1R转基因,CD11,TIM4,LAMP1和MHC II的表达,肺中MNP的亚群。覆盖支气管相关淋巴样组织的专门上皮细胞或M细胞表达CSF1R转基因,II型肺细胞表达LAMP1,表明这些上皮细胞是吞噬和转吞细胞抗原。从第4周开始,在气管中可见高度组织化的淋巴样组织。在各个年龄段的整个气囊中,CSF1R-transgene +细胞散落,而在以后的阶段,CSF1R-transgene +细胞内衬毛细血管。这些结果将为进一步鉴定巨噬细胞和树突状细胞及其在呼吸系统疾病和疫苗反应中的作用提供基础。

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