Abstract
African swine fever virus (ASFV) causes hemorrhagic disease in domestic pigs by replicating mainly in monocyte/macrophage lineages. Various primary cells including pulmonary alveolar macrophages have been used for the propagation of ASFV on this account. However, ethical constraints and consistency problems exist as it is necessary to harvest same phenotype of primary cells in order to continue a study. We suggested renal-derived swine macrophages as a novel primary cell candidate to address these issues. These primary cells proved to be permissive to both cell adapted ASFV and a wild-type ASFV. Compared to the commercial cell line MA-104, the renal-derived macrophages were more suitable to isolate the field virus. The consistent molecular characteristics of the renal-derived macrophages were demonstrated by immunocytochemistry with antibodies against macrophage cell surface markers including CD163, CD172a, and Iba-1. Viral protein p30 and p72 expression in ASFV infected macrophages was confirmed by immunocytochemistry by use of specific monoclonal antibodies. We observed increase of cell-free viral DNA and infectious virus titer in infected cell supernatant in successive days-post-infection. These results demonstrated that primary renal-derived swine macrophages are useful for ASFV isolation and propagation in terms of cell phenotypes, susceptibility to the virus, and virus production.
Highlights
African swine fever virus (ASFV) is an enveloped icosahedral double stranded DNA virus which belongs to the genus Asfivirus, family Asfaviridae and order incertae sedis [1]
Both macrophage monolayers infected with virus isolate and infectious tissue homogenates remained free of cytopathic effects until 7 dpi
A novel method of ASFV cultures on primary renalderived swine macrophages showing molecular and morphologic characteristics of monocyte/macrophage lineage was suggested with high continuity and reproducibility
Summary
African swine fever virus (ASFV) is an enveloped icosahedral double stranded DNA virus which belongs to the genus Asfivirus, family Asfaviridae and order incertae sedis [1]. Heterogeneity of ASFV Susceptibility in Renal-Derived Macrophages macrophage cell populations affecting permissiveness to virus infection is explained by molecular characteristics such as the virus specific membrane markers which are necessary for virus attachment and entry [8, 9]. It is well-known that ASFV replicates in various macrophage populations of spleen, lymph node, lung, liver and kidney [10,11,12]. Among various types of macrophages assessed, pulmonary alveolar macrophages (PAMs) were suggested as they are more susceptible to ASFV infection in comparison with bone marrow derived macrophages or blood monocytes
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
