Abstract
Foot-and-mouth disease virus (FMDV) causes a highly contagious disease with catastrophic economic impact for affected countries. BHK21 suspension cells are preferred for the industrial production of FMDV vaccine antigen, but not all virus strains can be successfully propagated in these cells. Serotype Asia-1 is often affected by this phenomenon. In this study, the Asia-1 strain Shamir was used to examine viral, cellular and environmental factors that contribute to resistance to cell culture infection. Cell media composition, pH and ammonium chloride concentration did not affect Asia-1 differently than other serotypes. Virus replication after transfection of viral genome was not impaired, but the adhesion to the cells was markedly reduced for Asia-1 in comparison to serotype A. The Asia-1 Shamir virus was successfully adapted to grow in the resistant cells by using a closely related but susceptible cell line. Sequence analysis of the adapted virus revealed two distinct mutations in the capsid protein VP1 that might mediate cell attachment and entry.
Highlights
In spite of international control efforts, foot-and-mouth disease virus (FMDV) is still widespread in the Middle East, Asia and Africa, where it causes severe disruptions of livestock production and trade [1]
BHK21 cells have some phenotypic features that are detrimental for production of FMDV antigen
Along with alterations in cell ploidy and a down-regulated surface expression of particular integrins correlated with the loss of actin stress fibers [6,7,8], BHK cells vary in their susceptibility for different FMDV strains [9,10]
Summary
In spite of international control efforts, foot-and-mouth disease virus (FMDV) is still widespread in the Middle East, Asia and Africa, where it causes severe disruptions of livestock production and trade [1]. Territories (SAT) 1–3 and Asia-1, and a previous infection with one serotype does not protect against an infection with any of the other six [2]. To fight this highly contagious disease, inactivated vaccines are produced at a large scale. The dominant cell line for industrial production of FMDV vaccine antigen is baby hamster kidney-21, clone 13 (BHK21C13) by MacPherson and Stoker [3], adapted to grow in suspension by Capstick et al [4], and used in large fermenters as first described by Telling and Elsworth [5]. The ability for FMDV infection can get lost on repeated subculturing [9,11]
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