Del Cacho Emilio, Gallego Margarita, Lillehoj Hyun S, López-Bernard Fernando, Sánchez-Acedo Caridad
Department of Animal Pathology, Faculty of Veterinary Sciences, University of Zaragoza, Zaragoza, Spain.
Vet Immunol Immunopathol. 2009 May 15;129(1-2):66-75. doi: 10.1016/j.vetimm.2008.12.015. Epub 2008 Dec 11.
An antiserum against Eimeria tenella sporozoites was used to localize and isolate Ag-binding cells in intestinal cecal tonsils of parasite-infected chickens. Based on their tissue localization, ultrastructural features, and expression of surface markers, two subpopulations of cells were isolated, CD45(+) interdigitating dendritic cells (IDCs) and CD45(-) follicular dendritic cells (FDCs). IDCs expressed MHC class I, MHC class II, and selectin, induced the proliferation of allogeneic naïve CD4(+) T cells, and increased the secretion of IFN-gamma by autologous T cells. FDCs expressed surface IgG, IgM, ICAM-1, and VCAM-1, stimulated the proliferation of LPS-treated allogeneic B cells, and augmented the secretion of IgG by LPS-treated autologous B cells. Final cell yields were 6 x 10(5) to 8 x 10(5) cells per chicken with >95% purity. In summary, this combination of methods using Abs against E. tenella and CD45 made it possible for the first time to obtain a highly enriched IDCs and FDCs which are functionally active in chickens. This novel method will enable the detailed biochemical and immunological characterizations of avian dendritic cells and facilitate the investigation of their role in initiating immune response in normal and disease states.
使用抗柔嫩艾美耳球虫子孢子的抗血清,在感染寄生虫的鸡的肠道盲肠扁桃体中定位并分离出抗原结合细胞。根据它们的组织定位、超微结构特征和表面标志物的表达,分离出两个细胞亚群,即CD45(+) 交错突细胞(IDCs)和CD45(-) 滤泡树突状细胞(FDCs)。IDCs表达MHC I类、MHC II类和选择素,诱导同种异体幼稚CD4(+) T细胞增殖,并增加自体T细胞分泌IFN-γ。FDCs表达表面IgG、IgM、ICAM-1和VCAM-1,刺激经LPS处理的同种异体B细胞增殖,并增加经LPS处理的自体B细胞分泌IgG。每只鸡的最终细胞产量为6×10(5) 至8×10(5) 个细胞,纯度>95%。总之,这种使用抗柔嫩艾美耳球虫抗体和CD45的方法组合,首次使得获得在鸡中具有功能活性的高度富集的IDCs和FDCs成为可能。这种新方法将能够对禽类树突状细胞进行详细的生化和免疫学表征,并有助于研究它们在正常和疾病状态下启动免疫反应中的作用。