Abstract
Increasing clinical lines of evidence have shown the coinfection/superinfection of porcine circovirus type 2 (PCV2) and classical swine fever virus (CSFV). Here, we investigated whether PCV2 and CSFV could infect the same cell productively by constructing an in vitro coinfection model. Our results indicated that PCV2-free PK15 cells but not ST cells were more sensitive to PCV2, and the PK15 cell line could stably harbor replicating CSFV (PK15-CSFV cells) with a high infection rate. Confocal and super-resolution microscopic analysis showed that PCV2 and CSFV colocalized in the same PK15-CSFV cell, and the CSFV E2 protein translocated from the cytoplasm to the nucleus in PK15-CSFV cells infected with PCV2. Moreover, PCV2-CSFV dual-positive cells increased gradually in PK15-CSFV cells in a PCV2 dose-dependent manner. In PK15-CSFV cells, PCV2 replicated well, and the production of PCV2 progeny was not influenced by CSFV infection. However, CSFV reproduction decreased in a PCV2 dose-dependent manner. In addition, cellular apoptosis was not strengthened in PK15-CSFV cells infected with PCV2 in comparison with PCV2-infected PK15 cells. Moreover, using this coinfection model we further demonstrated PCV2-induced apoptosis might contribute to the impairment of CSFV HCLV strain replication in coinfected cells. Taken together, our results demonstrate for the first time the coinfection/superinfection of PCV2 and CSFV within the same cell, providing an in vitro model to facilitate further investigation of the underlying mechanism of CSFV and PCV2 coinfection.
Highlights
Virus coinfection or superinfection, a simultaneous or consecutive infection, has become a common phenomenon which involves the infection of the same type virus, a closely related virus or different virus species
After the cells were infected with porcine circovirus type 2 (PCV2) at the multiplicity of infection (MOI), average percentages of PCV2-positive cells were counted by selecting four random fields of view of the tested cells, and progeny virus titers were determined in PK15 cells
These results showed that the PK15 cell line was the most permissive for PCV2 and could be infected in a PCV2 dose-dependent manner
Summary
Thereby, we aimed to establish a coinfection system of CSFV and PCV2 in vitro, and provide an efficient way to study the potential underlying mechanism for CSFV and PCV2 coinfection and other virus coinfection, as well
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