MRC-University of Glasgow Centre for Virus Research, Glasgow, United Kingdom.
Moredun Research Institute, Penicuik, United Kingdom.
Front Immunol. 2018 Dec 11;9:2892. doi: 10.3389/fimmu.2018.02892. eCollection 2018.
Sheep are not only a major livestock species globally, they are also an important large animal model for biomedical research and have contributed to our understanding of the ontogeny and architecture of the mammalian immune system. In this study, we applied immunohistochemistry and multicolor immunofluorescence in fixed and paraffin-embedded lymph nodes to phenotype the key populations of antigen presenting cells, lymphocytes, and stromal cells that orchestrate the host adaptive immune response. We used an extensive panel of antibodies directed against markers associated with dendritic cells (MHC class II, CD83, and CD208), macrophages (CD11b, CD163, and CD169), stromal cells (CNA.42, S100, and CD83), and lymphocytes (CD3, Pax5, CD4, CD8). Using different methods of tissue fixation and antigen retrieval, we provide a detailed immunophenotyping of sheep lymph nodes including the identification of potential subpopulations of antigen presenting cells and stromal cells. By characterizing cells expressing combinations of these markers in the context of their morphology and location within the lymph node architecture, we provide valuable new tools to investigate the structure, activation, and regulation of the sheep immune system in health and disease.
绵羊不仅是全球重要的家畜物种，也是生物医学研究中重要的大型动物模型，为我们理解哺乳动物免疫系统的个体发生和结构做出了贡献。在这项研究中，我们应用免疫组织化学和固定石蜡包埋淋巴结的多色免疫荧光技术，表型分析了主导宿主适应性免疫反应的抗原呈递细胞、淋巴细胞和基质细胞的关键群体。我们使用了针对与树突状细胞（MHC 类 II、CD83 和 CD208）、巨噬细胞（CD11b、CD163 和 CD169）、基质细胞（CNA.42、S100 和 CD83）和淋巴细胞（CD3、Pax5、CD4 和 CD8）相关标记物的广泛抗体面板。通过使用不同的组织固定和抗原修复方法，我们对绵羊淋巴结进行了详细的免疫表型分析，包括鉴定潜在的抗原呈递细胞和基质细胞亚群。通过对在淋巴结结构中形态和位置表达这些标记物组合的细胞进行特征描述，我们为研究健康和疾病状态下绵羊免疫系统的结构、激活和调节提供了有价值的新工具。
