Abstract:
Objective Feasibility of using mouse macrophage cell line Raw 264.7 for studies on pathogenesis of Mycoplasma ovipneumoniae(Mo) on sheep was investigated by comparing the proteome changes taken place in both macrophages after an induced Mo infection.
Method The primary alveolar macrophages from sheep lung tissue were isolated and used, along with Raw 264.7, for the study. Both macrophages were infected with a standard strain of Mo (MOI=10) for 24h prior to proteomic and RT-PCR analyses to compare the expressions on the target genes in them.
Result The image of specific surface antigen CD14 shown on the cells obtained from the lung tissue confirmed the identity of the isolated sheep primary alveolar macrophages. After infected by Mo, both sheep and mouse macrophages showed a similar significant change pattern on the expressions of FADD, IL-1β, NOS2, and THBS1. These proteins are known to associated with the Toll-like receptor signaling pathway, MAPK signaling pathway, and autophagy in biological processes.
Conclusion It appeared that Raw 264.7 could be satisfactorily used to substitute the sheep primary alveolar macrophage in a simulated system for studies on Mo pathogenesis to considerably simplify the process and save cost in conducting the experiments on sheep.
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