a Department of Pathology , University of Georgia , Athens , GA , USA.
b Department of Veterinary Microbiology and Pathology , Washington State University , Pullman , WA , USA.
Vet Q. 2017 Dec;37(1):52-56. doi: 10.1080/01652176.2017.1297550.
Ruminants, including sheep and goats (small ruminants), are key agricultural animals in many parts of the world. Infectious diseases, including many viral diseases, are significant problems to efficient production of ruminants. Unfortunately, reagents tailored to viruses of ruminants, and especially small ruminants, are lacking compared to other animals more typically used for biomedical research.
The purpose of this study was to determine the permissibility of a stably immortalized, sheep microglial cell line to viruses that are reported to infect ruminants: bovine viral diarrhea virus (BVDV), bovine herpesvirus 1 (BoHV-1), small ruminant lentiviruses (SRLV), and bovine respiratory syncytial virus (BRSV).
Sublines A and H of previously isolated, immortalized, and characterized (CD14-positive) ovine microglial cells were used. Bovine turbinate cells and goat synovial membrane cells were used for comparison. Cytopathic changes were used to confirm infection of individual wells, which were then counted and used to calculate the 50% tissue culture infectious dose. Uninoculated cells served as negative controls and confirmed that the cells were not previously infected with these viruses using polymerase chain reaction (PCR).
Inoculation of the two microglial cell sublines with laboratory and field isolates of BVDV, BoHV-1, and BRSV resulted in viral infection in a manner similar to bovine turbinate cells. Immortalized microglia cells are also permissive to SRLV, similar to goat synovial membrane cells.
These immortalized sheep microglial cells provide a new tool for the study of ruminant viruses in ruminant microglial cell line.
反刍动物,包括绵羊和山羊(小反刍动物),是世界许多地区的重要农业动物。传染病,包括许多病毒性疾病,是反刍动物高效生产的重大问题。不幸的是,与其他更常用于生物医学研究的动物相比,针对反刍动物,尤其是小反刍动物的病毒的试剂相对缺乏。
本研究旨在确定一种稳定永生化的绵羊小胶质细胞系对感染反刍动物的病毒的允许性:牛病毒性腹泻病毒(BVDV)、牛疱疹病毒 1(BoHV-1)、小反刍动物慢病毒(SRLV)和牛呼吸道合胞体病毒(BRSV)。
使用先前分离、永生化和表征(CD14 阳性)的绵羊小胶质细胞的亚系 A 和 H。牛鼻甲细胞和山羊滑膜膜细胞用于比较。细胞病变作用用于确认单个孔的感染,然后对其进行计数并用于计算 50%组织培养感染剂量。未接种的细胞作为阴性对照,并使用聚合酶链反应(PCR)确认这些细胞先前未感染这些病毒。
用实验室和现场分离株 BVDV、BoHV-1 和 BRSV 接种两种小胶质细胞亚系,导致病毒感染类似于牛鼻甲细胞。永生化小胶质细胞也允许 SRLV 感染,类似于山羊滑膜膜细胞。
这些永生化的绵羊小胶质细胞为研究反刍动物小胶质细胞系中的反刍动物病毒提供了一种新工具。