Abstract
Virus transmission is essential for spreading viral infections and is a highly coordinated process which occurs by cell-free transmission or cell–cell contact. The transmission of Bovine Foamy Virus (BFV) is highly cell-associated, with undetectable cell-free transmission. However, BFV particle budding can be induced by overexpression of wild-type (wt) BFV Gag and Env or artificial retargeting of Gag to the plasma membrane via myristoylation membrane targeting signals, closely resembling observations in other foamy viruses. Thus, the particle release machinery of wt BFV appears to be an excellent model system to study viral adaption to cell-free transmission by in vitro selection and evolution. Using selection for BFV variants with high cell-free infectivity in bovine and non-bovine cells, infectivity dramatically increased from almost no infectious units to about 105–106 FFU (fluorescent focus forming units)/mL in both cell types. Importantly, the selected BFV variants with high titer (HT) cell-free infectivity could still transmit via cell-cell contacts and were neutralized by serum from naturally infected cows. These selected HT–BFV variants will shed light into virus transmission and potential routes of intervention in the spread of viral infections. It will also allow the improvement or development of new promising approaches for antiretroviral therapies.
Highlights
Spumaretroviruses, known as Foamy Viruses (FVs), were initially discovered in 1954 by Enders and Peebles in primary monkey kidney cell cultures [1]
The baby hamster kidney cell line BHK-21 (ATCC CCL-10) and Madin-Darby bovine kidney (MDBK) cells were grown in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal horse serum (FHS) and 1% penicillin-streptomycin solution and used for cell-free Bovine Foamy Virus (BFV) serial passaging and selection
The BFV indicator cell line (BICL) and MDBK indicator cell line (MICL) for BFV were derived from BHK and MDBK cells, respectively, and contained a green fluorescent protein (Gfp) expression cassette under the control of the BFV long-terminal repeat (LTR) promoter [37]
Summary
Spumaretroviruses, known as Foamy Viruses (FVs), were initially discovered in 1954 by Enders and Peebles in primary monkey kidney cell cultures [1]. FVs release non-infectious Env-only subviral particles and there is a strict dependence on capsid-glycoprotein interactions for virion release from the cells [5,6,7]. These and other unique features of FVs may be related to their unconventional gene expression and replication strategies, and a long FV-host co-evolution [2,8]. Due to the apparent lack of pathogenicity and their broad tissue tropism, FVs are promising vectors for gene and vaccine antigen delivery [5]
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