1887

Abstract

Varicella-zoster virus (VZV), the causative agent of chickenpox and herpes zoster, can be life-threatening in prematurely born children and in children with immune defects or who are under immunosuppressive treatment. Therefore agents for passive immunization, such as VZV-specific immunoglobulin preparations (VZIG) derived from convalescent plasma, are crucial in the prophylaxis of VZV infection. This study describes the isolation of human VZV-neutralizing recombinant antibodies. A human single-chain variable fragment (scFv) phage display library was generated from RNA extracted from peripheral blood lymphocytes of a convalescent varicella patient. Specific phage antibodies were selected against VZV-infected human fibroblasts, and eight unique clones were further expressed as soluble scFv in . They all showed binding characteristics to varicella antigens with affinities in the range 0·1–0·2 μM. Two of the scFv antibodies, VZV4 and VZV5, showed dose-dependent neutralization of VZV. VZV39 also showed a neutralizing effect as scFv, an effect that was increased 4000-fold by conversion into IgG and was further increased by the addition of complement. This is possibly the first time that monovalent scFv antibodies have been shown to neutralize VZV . This finding will have an impact on the production of new prophylactic antibodies, as such antibody fragments can be cost-effectively produced in . The antibodies isolated bind both complement-dependent and -independent epitopes for neutralization, thus they may prove useful tools for the study of VZV virulence mechanisms.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.80406-0
2004-12-01
2019-11-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/85/12/vir853493.html?itemId=/content/journal/jgv/10.1099/vir.0.80406-0&mimeType=html&fmt=ahah

References

  1. Boyum, A. ( 1968; ). Separation of leukocytes from blood and bone marrow. Introduction. Scand J Clin Lab Invest 21, S97.
    [Google Scholar]
  2. Brekke, O. H. & Sandlie, I. ( 2003; ). Therapeutic antibodies for human diseases at the dawn of the twenty-first century. Nat Rev Drug Discov 2, 52–62.[CrossRef]
    [Google Scholar]
  3. Brekke, O. H., Michaelsen, T. E., Sandin, R. & Sandlie, I. ( 1993; ). Activation of complement by an IgG molecule without a genetic hinge. Nature 363, 628–630.[CrossRef]
    [Google Scholar]
  4. Davison, A. J. & Scott, J. E. ( 1986; ). The complete DNA sequence of varicella-zoster virus. J Gen Virol 67, 1759–1816.[CrossRef]
    [Google Scholar]
  5. de Haard, H. J., van Neer, N., Reurs, A., Hufton, S. E., Roovers, R. C., Henderikx, P., de Bruine, A. P., Arends, J. W. & Hoogenboom, H. R. ( 1999; ). A large non-immunized human Fab fragment phage library that permits rapid isolation and kinetic analysis of high affinity antibodies. J Biol Chem 274, 18218–18230.[CrossRef]
    [Google Scholar]
  6. Drew, P. D., Moss, M. T., Pasieka, T. J., Grose, C., Harris, W. J. & Porter, A. J. ( 2001; ). Multimeric humanized varicella-zoster virus antibody fragments to gH neutralize virus while monomeric fragments do not. J Gen Virol 82, 1959–1963.
    [Google Scholar]
  7. Foung, S. K., Perkins, S., Koropchak, C., Fishwild, D. M., Wittek, A. E., Engleman, E. G., Grumet, F. C. & Arvin, A. M. ( 1985; ). Human monoclonal antibodies neutralizing varicella-zoster virus. J Infect Dis 152, 280–285.[CrossRef]
    [Google Scholar]
  8. Griffiths, A. D., Williams, S. C., Hartley, O. & 15 other authors ( 1994; ). Isolation of high affinity human antibodies directly from large synthetic repertoires. EMBO J 13, 3245–3260.
    [Google Scholar]
  9. Grose, C. ( 1990; ). Glycoproteins encoded by Varicella-zoster virus: biosynthesis, phosphorylation, and intracellular trafficking. Annu Rev Microbiol 44, 59–80.[CrossRef]
    [Google Scholar]
  10. Knappik, A., Ge, L., Honegger, A. & 7 other authors ( 2000; ). Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides. J Mol Biol 296, 57–86.[CrossRef]
    [Google Scholar]
  11. Le Gall, F., Reusch, U., Moldenhauer, G., Little, M. & Kipriyanov, S. M. ( 2004; ). Immunosuppressive properties of anti-CD3 single-chain Fv and diabody. J Immunol Methods 285, 111–127.[CrossRef]
    [Google Scholar]
  12. Little, M., Welschof, M., Braunagel, M. & 8 other authors ( 1999; ). Generation of a large complex antibody library from multiple donors. J Immunol Methods 231, 3–9.[CrossRef]
    [Google Scholar]
  13. Lloyd-Evans, P. & Gilmour, J. E. ( 2000; ). Expression of neutralizing recombinant human antibodies against varicella zoster virus for use as a potential prophylactic. Hybridoma 19, 143–149.[CrossRef]
    [Google Scholar]
  14. Michaelsen, T. E., Aase, A., Westby, C. & Sandlie, I. ( 1990; ). Enhancement of complement activation and cytolysis of human IgG3 by deletion of hinge exons. Scand J Immunol 32, 517–528.[CrossRef]
    [Google Scholar]
  15. Montalvo, E. A. & Grose, C. ( 1986; ). Neutralization epitope of varicella zoster virus on native viral glycoprotein gp118 (VZV glycoprotein gpIII). Virology 149, 230–241.[CrossRef]
    [Google Scholar]
  16. Norderhaug, L., Olafsen, T., Michaelsen, T. E. & Sandlie, I. ( 1997; ). Versatile vectors for transient and stable expression of recombinant antibody molecules in mammalian cells. J Immunol Methods 204, 77–87.[CrossRef]
    [Google Scholar]
  17. Sblattero, D. & Bradbury, A. ( 1998; ). A definitive set of oligonucleotide primers for amplifying human V regions. Immunotechnology 3, 271–278.[CrossRef]
    [Google Scholar]
  18. Sblattero, D. & Bradbury, A. ( 2000; ). Exploiting recombination in single bacteria to make large phage antibody libraries. Nat Biotechnol 18, 75–80.[CrossRef]
    [Google Scholar]
  19. Sheets, M. D., Amersdorfer, P., Finnern, R. & 8 other authors ( 1998; ). Efficient construction of a large nonimmune phage antibody library: the production of high-affinity human single-chain antibodies to protein antigens. Proc Natl Acad Sci U S A 95, 6157–6162.[CrossRef]
    [Google Scholar]
  20. Soderlind, E., Strandberg, L., Jirholt, P. & 10 other authors ( 2000; ). Recombining germline-derived CDR sequences for creating diverse single-framework antibody libraries. Nat Biotechnol 18, 852–856.[CrossRef]
    [Google Scholar]
  21. Stacy, J. E., Kausmally, L., Simonsen, B., Nordgard, S. H., Alsoe, L., Michaelsen, T. E. & Brekke, O. H. ( 2003; ). Direct isolation of recombinant human antibodies against group B Neisseria meningitidis from scFv expression libraries. J Immunol Methods 283, 247–259.[CrossRef]
    [Google Scholar]
  22. Sugano, T., Matsumoto, Y., Miyamoto, C. & Masuho, Y. ( 1987; ). Hybridomas producing human monoclonal antibodies against varicella-zoster virus. Eur J Immunol 17, 359–364.[CrossRef]
    [Google Scholar]
  23. Sui, J., Li, W., Murakami, A. & 11 other authors ( 2004; ). Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association. Proc Natl Acad Sci U S A 101, 2536–2541.[CrossRef]
    [Google Scholar]
  24. Vaughan, T. J., Williams, A. J., Pritchard, K. & 7 other authors ( 1996; ). Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library. Nat Biotechnol 14, 309–314.[CrossRef]
    [Google Scholar]
  25. Williamson, R. A., Burioni, R., Sanna, P. P., Partridge, L. J., Barbas, C. F., III & Burton, D. R. ( 1993; ). Human monoclonal antibodies against a plethora of viral pathogens from single combinatorial libraries. Proc Natl Acad Sci U S A 90, 4141–4145. [Erratum, Proc Natl Acad Sci U S A (1994) 91, 1193.]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.80406-0
Loading
/content/journal/jgv/10.1099/vir.0.80406-0
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error