Propagation of an attenuated virus by design: engineering a novel receptor for a noninfectious foot-and-mouth disease virus.
To gain entry into cells, viruses utilize a variety of different cell-surface molecules. Foot-and-mouth disease virus (FMDV) binds to cell-surface integrin molecules via an arginine-glycine-aspartic acid (RGD) sequence in capsid protein VP1. Binding to this particular cell-surface molecule influences FMDV tropism, and virus/receptor interactions appear to be responsible, in part, for selection of antigenic variants. To study early events of virus-cell interaction, we engineered an alternative and novel receptor for FMDV. Specifically, we generated a new receptor by fusing a virus-binding, single-chain antibody (scAb) to intracellular adhesion molecule 1 (ICAM1). Cells that are normally not susceptible to FMDV infection became susceptible after being transfected with DNA encoding the scAb/ICAM1 protein. An escape mutant (B2PD.3), derived with the mAb used to generate the genetically engineered receptor, was restricted for growth on the scAb/ICAM1 cells, but a variant of B2PD.3 selected by propagation on scAb/ICAM1 cells grew well on these cells. This variant partially regained wild-type sequence in the epitope recognized by the mAb and also regained the ability to be neutralize by the mAb. Moreover, RGD-deleted virions that are noninfectious in animals and other cell types grew to high titers and were able to form plaques on scAb/ ICAM1 cells. These studies demonstrate the first production of a totally synthetic cell-surface receptor for a virus. This novel approach will be useful for studying virus reception and for the development of safer vaccines against viral pathogens of animals and humans.
Rieder
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E
Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY 11944, USA.
Berinstein
A
A
Baxt
B
B
Kang
A
A
Mason
P W
PW
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D005537:Aphthovirus / Q000502:physiology*; D001665:Binding Sites; D016466:CHO Cells; D002213:Capsid / Q000096:biosynthesis / Q000502:physiology*; D036022:Capsid Proteins; D002460:Cell Line; D006224:Cricetinae; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009842:Oligopeptides; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015202:Protein Engineering; D011991:Receptors, Virus / Q000096:biosynthesis / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D014162:Transfection; D010948:Viral Plaque Assay
D000914:Antibodies, Viral; D036022:Capsid Proteins; D009842:Oligopeptides; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; C442501:VP1 protein, Foot-and-mouth disease virus; D018799:Intercellular Adhesion Molecule-1; C047981:arginyl-glycyl-aspartic acid