Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Sep 17, 1996; 93(19): 10428–10433.

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.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.4M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Mendelsohn CL, Wimmer E, Racaniello VR. Cellular receptor for poliovirus: molecular cloning, nucleotide sequence, and expression of a new member of the immunoglobulin superfamily. Cell. 1989 Mar 10;56(5):855–865. [PubMed]
  • Greve JM, Davis G, Meyer AM, Forte CP, Yost SC, Marlor CW, Kamarck ME, McClelland A. The major human rhinovirus receptor is ICAM-1. Cell. 1989 Mar 10;56(5):839–847. [PubMed]
  • Staunton DE, Merluzzi VJ, Rothlein R, Barton R, Marlin SD, Springer TA. A cell adhesion molecule, ICAM-1, is the major surface receptor for rhinoviruses. Cell. 1989 Mar 10;56(5):849–853. [PubMed]
  • Tomassini JE, Graham D, DeWitt CM, Lineberger DW, Rodkey JA, Colonno RJ. cDNA cloning reveals that the major group rhinovirus receptor on HeLa cells is intercellular adhesion molecule 1. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4907–4911. [PMC free article] [PubMed]
  • Hofer F, Gruenberger M, Kowalski H, Machat H, Huettinger M, Kuechler E, Blaas D. Members of the low density lipoprotein receptor family mediate cell entry of a minor-group common cold virus. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1839–1842. [PMC free article] [PubMed]
  • Bergelson JM, Shepley MP, Chan BM, Hemler ME, Finberg RW. Identification of the integrin VLA-2 as a receptor for echovirus 1. Science. 1992 Mar 27;255(5052):1718–1720. [PubMed]
  • Roivainen M, Piirainen L, Hovi T, Virtanen I, Riikonen T, Heino J, Hyypiä T. Entry of coxsackievirus A9 into host cells: specific interactions with alpha v beta 3 integrin, the vitronectin receptor. Virology. 1994 Sep;203(2):357–365. [PubMed]
  • Berinstein A, Roivainen M, Hovi T, Mason PW, Baxt B. Antibodies to the vitronectin receptor (integrin alpha V beta 3) inhibit binding and infection of foot-and-mouth disease virus to cultured cells. J Virol. 1995 Apr;69(4):2664–2666. [PMC free article] [PubMed]
  • Mason PW, Rieder E, Baxt B. RGD sequence of foot-and-mouth disease virus is essential for infecting cells via the natural receptor but can be bypassed by an antibody-dependent enhancement pathway. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1932–1936. [PMC free article] [PubMed]
  • McKenna TS, Lubroth J, Rieder E, Baxt B, Mason PW. Receptor binding site-deleted foot-and-mouth disease (FMD) virus protects cattle from FMD. J Virol. 1995 Sep;69(9):5787–5790. [PMC free article] [PubMed]
  • Peiris JS, Gordon S, Unkeless JC, Porterfield JS. Monoclonal anti-Fc receptor IgG blocks antibody enhancement of viral replication in macrophages. Nature. 1981 Jan 15;289(5794):189–191. [PubMed]
  • Schlesinger JJ, Brandriss MW. Growth of 17D yellow fever virus in a macrophage-like cell line, U937: role of Fc and viral receptors in antibody-mediated infection. J Immunol. 1981 Aug;127(2):659–665. [PubMed]
  • Burstin SJ, Brandriss MW, Schlesinger JJ. Infection of a macrophage-like cell line, P388D1 with reovirus; effects of immune ascitic fluids and monoclonal antibodies on neutralization and on enhancement of viral growth. J Immunol. 1983 Jun;130(6):2915–2919. [PubMed]
  • Halstead SB. Pathogenesis of dengue: challenges to molecular biology. Science. 1988 Jan 29;239(4839):476–481. [PubMed]
  • Mason PW, Baxt B, Brown F, Harber J, Murdin A, Wimmer E. Antibody-complexed foot-and-mouth disease virus, but not poliovirus, can infect normally insusceptible cells via the Fc receptor. Virology. 1993 Feb;192(2):568–577. [PubMed]
  • Baxt B, Mason PW. Foot-and-mouth disease virus undergoes restricted replication in macrophage cell cultures following Fc receptor-mediated adsorption. Virology. 1995 Mar 10;207(2):503–509. [PubMed]
  • Salt JS. The carrier state in foot and mouth disease--an immunological review. Br Vet J. 1993 May-Jun;149(3):207–223. [PubMed]
  • Kaplan G, Freistadt MS, Racaniello VR. Neutralization of poliovirus by cell receptors expressed in insect cells. J Virol. 1990 Oct;64(10):4697–4702. [PMC free article] [PubMed]
  • Greve JM, Forte CP, Marlor CW, Meyer AM, Hoover-Litty H, Wunderlich D, McClelland A. Mechanisms of receptor-mediated rhinovirus neutralization defined by two soluble forms of ICAM-1. J Virol. 1991 Nov;65(11):6015–6023. [PMC free article] [PubMed]
  • Selinka HC, Zibert A, Wimmer E. Poliovirus can enter and infect mammalian cells by way of an intercellular adhesion molecule 1 pathway. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3598–3602. [PMC free article] [PubMed]
  • Bernhardt G, Harber J, Zibert A, deCrombrugghe M, Wimmer E. The poliovirus receptor: identification of domains and amino acid residues critical for virus binding. Virology. 1994 Sep;203(2):344–356. [PubMed]
  • Selinka HC, Zibert A, Wimmer E. A chimeric poliovirus/CD4 receptor confers susceptibility to poliovirus on mouse cells. J Virol. 1992 Apr;66(4):2523–2526. [PMC free article] [PubMed]
  • Huse WD, Sastry L, Iverson SA, Kang AS, Alting-Mees M, Burton DR, Benkovic SJ, Lerner RA. Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda. Science. 1989 Dec 8;246(4935):1275–1281. [PubMed]
  • Riechmann L, Weill M, Cavanagh J. Improving the antigen affinity of an antibody Fv-fragment by protein design. J Mol Biol. 1992 Apr 20;224(4):913–918. [PubMed]
  • Lerner RA, Kang AS, Bain JD, Burton DR, Barbas CF., 3rd Antibodies without immunization. Science. 1992 Nov 20;258(5086):1313–1314. [PubMed]
  • He M, Kang AS, Hamon M, Humphreys AS, Gani M, Taussig MJ. Characterization of a progesterone-binding, three-domain antibody fragment (VH/K) expressed in Escherichia coli. Immunology. 1995 Apr;84(4):662–668. [PMC free article] [PubMed]
  • Kang AS, Jones TM, Burton DR. Antibody redesign by chain shuffling from random combinatorial immunoglobulin libraries. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11120–11123. [PMC free article] [PubMed]
  • Bird RE, Hardman KD, Jacobson JW, Johnson S, Kaufman BM, Lee SM, Lee T, Pope SH, Riordan GS, Whitlow M. Single-chain antigen-binding proteins. Science. 1988 Oct 21;242(4877):423–426. [PubMed]
  • Huston JS, Levinson D, Mudgett-Hunter M, Tai MS, Novotný J, Margolies MN, Ridge RJ, Bruccoleri RE, Haber E, Crea R, et al. Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5879–5883. [PMC free article] [PubMed]
  • Rieder E, Bunch T, Brown F, Mason PW. Genetically engineered foot-and-mouth disease viruses with poly(C) tracts of two nucleotides are virulent in mice. J Virol. 1993 Sep;67(9):5139–5145. [PMC free article] [PubMed]
  • Baxt B, Vakharia V, Moore DM, Franke AJ, Morgan DO. Analysis of neutralizing antigenic sites on the surface of type A12 foot-and-mouth disease virus. J Virol. 1989 May;63(5):2143–2151. [PMC free article] [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Staunton DE, Marlin SD, Stratowa C, Dustin ML, Springer TA. Primary structure of ICAM-1 demonstrates interaction between members of the immunoglobulin and integrin supergene families. Cell. 1988 Mar 25;52(6):925–933. [PubMed]
  • Haywood AM. Virus receptors: binding, adhesion strengthening, and changes in viral structure. J Virol. 1994 Jan;68(1):1–5. [PMC free article] [PubMed]
  • Staunton DE, Dustin ML, Erickson HP, Springer TA. The arrangement of the immunoglobulin-like domains of ICAM-1 and the binding sites for LFA-1 and rhinovirus. Cell. 1990 Apr 20;61(2):243–254. [PubMed]
  • Domingo E, Díez J, Martínez MA, Hernández J, Holguín A, Borrego B, Mateu MG. New observations on antigenic diversification of RNA viruses. Antigenic variation is not dependent on immune selection. J Gen Virol. 1993 Oct;74(Pt 10):2039–2045. [PubMed]
  • Rieder E, Baxt B, Mason PW. Animal-derived antigenic variants of foot-and-mouth disease virus type A12 have low affinity for cells in culture. J Virol. 1994 Aug;68(8):5296–5299. [PMC free article] [PubMed]
  • Colston EM, Racaniello VR. Poliovirus variants selected on mutant receptor-expressing cells identify capsid residues that expand receptor recognition. J Virol. 1995 Aug;69(8):4823–4829. [PMC free article] [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...