Antibody-targeted cell fusion

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Membrane fusion has many potential applications in biotechnology. Here we show that antibody-targeted cell fusion can be achieved by engineering a fusogenic viral membrane glycoprotein complex. Three different single-chain antibodies were displayed at the extracellular C terminus of the measles hemagglutinin (H) protein, and combinations of point mutations were introduced to ablate its ability to trigger fusion through the native viral receptors CD46 and SLAM. When coexpressed with the measles fusion (F) protein, using plasmid cotransfection or bicistronic adenoviral vectors, the retargeted H proteins could mediate antibody-targeted cell fusion of receptor-negative or receptor-positive index cells with receptor-positive target cells. Adenoviral expression vectors mediating human epidermal growth factor receptor (EGFR)-targeted cell fusion were potently cytotoxic against EGFR-positive tumor cell lines and showed superior antitumor potency against EGFR-positive tumor xenografts as compared with control adenoviruses expressing native (untargeted) or CD38-targeted H proteins.

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Figure 1: Fusogenic properties of mutant H glycoproteins displaying anti-CD38 antibody.
Figure 2: Antibody-targeted cell fusion and cell killing.
Figure 3: Targeted cytoreductive gene therapy using homologous targeted cell fusion.
Figure 4: Adenoviral vectors mediating homologous or heterologous targeted cell fusion.


  1. 1

    Shemer, G. & Podbilewicz, B. Fusomorphogenesis: cell fusion in organ formation. Dev. Dyn. 218, 30–51 (2000).

  2. 2

    Holden, C. & Vogel, G. Stem cells. Plasticity: time for a reappraisal. Science 296, 2126–2129 (2002).

  3. 3

    Griffin, D.E. Measles Virus. in Fields Virology (eds. Knipe, D.M. & Howley, P.M.) 1402–1442 (Lippincott Williams & Wilkins, Philadelphia, 2001).

  4. 4

    Freed, E.O. & Martin, M.A. HIVs and their replication. in Fields Virology (eds. Knipe, D.M. & Howley, P.M.) 1971–2042 (Lippincott Williams & Wilkins, Philadelphia, 2001).

  5. 5

    Cohen, J.I. & Straus, S.E. Varicella-Zoster virus and its replication. in Fields Virology (eds. Knipe, D.M. & Howley, P.M.) 2707–2730 (Lippincott Williams & Wilkins, Philadelphia, 2001).

  6. 6

    Kohler, G. & Milstein, C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256, 495–497 (1975).

  7. 7

    Dorssers, L.C. & Veldscholte, J. Identification of a novel breast-cancer-anti-estrogen-resistance (BCAR2) locus by cell-fusion-mediated gene transfer in human breast-cancer cells. Int. J. Cancer 72, 700–7005 (1997).

  8. 8

    Dieken, E.S., Epner, E.M., Fiering, S., Fournier, R.E. & Groudine, M. Efficient modification of human chromosomal alleles using recombination-proficient chicken/human microcell hybrids. Nat. Genet. 12, 174–182 (1996).

  9. 9

    Galanis, E. et al. Use of viral fusogenic membrane glycoproteins as novel therapeutic transgenes in gliomas. Hum. Gene Ther. 12, 811–821 (2001).

  10. 10

    Peng, K.W. et al. Intraperitoneal therapy of ovarian cancer using an engineered measles virus. Cancer Res. 62, 4656–4662 (2002).

  11. 11

    Guo, Y. et al. Effective tumor vaccine generated by fusion of hepatoma cells with activated B cells. Science 263, 518–520 (1994).

  12. 12

    Bateman, A. et al. Fusogenic membrane glycoproteins as a novel class of genes for the local and immune-mediated control of tumor growth. Cancer Res. 60, 1492–1497 (2000).

  13. 13

    Dorig, R.E., Marcil, A., Chopra, A. & Richardson, C.D. The human CD46 molecule is a receptor for measles virus (Edmonston strain). Cell 75, 295–305 (1993).

  14. 14

    Tatsuo, H., Ono, N., Tanaka, K. & Yanagi, Y. SLAM (CDw150) is a cellular receptor for measles virus. Nature 406, 893–897 (2000).

  15. 15

    von Messling, V., Zimmer, G., Herrler, G., Haas, L. & Cattaneo, R. The hemagglutinin of canine distemper virus determines tropism and cytopathogenicity. J. Virol. 75, 6418–6427 (2001).

  16. 16

    Hammond, A.L. et al. Single-chain antibody displayed on a recombinant measles virus confers entry through the tumor-associated carcinoembryonic antigen. J. Virol. 75, 2087–2097 (2001).

  17. 17

    Peng, K.W. et al. Oncolytic measles viruses displaying a single-chain antibody against CD38, a myeloma cell marker. Blood 101, 2557–2562 (2003).

  18. 18

    Lecouturier, V. et al. Identification of two amino acids in the hemagglutinin glycoprotein of measles virus (MV) that govern hemadsorption, HeLa cell fusion, and CD46 downregulation: phenotypic markers that differentiate vaccine and wild-type MV strains. J. Virol. 70, 4200–4204 (1996).

  19. 19

    Vongpunsawad, S., Oezgum, N., Braun, W. & Cattaneo, R. Selectively receptor-bind measles viruses: Identification of the SLAM-and CD46-interacting residues and their localization on a new hemagglutinin structural model. J. Virol. 78, 302–313 (2004).

  20. 20

    Xie, M., Tanaka, K., Ono, N., Minagawa, H. & Yanagi, Y. Amino acid substitutions at position 481 differently affect the ability of the measles virus hemagglutinin to induce cell fusion in monkey and marmoset cells co-expressing the fusion protein. Arch. Virol. 144, 1689–1699 (1999).

  21. 21

    Cattaneo, R. & Rose, J.K. Cell fusion by the envelope glycoproteins of persistent measles viruses which caused lethal human brain disease. J. Virol. 67, 1493–1502 (1993).

  22. 22

    Chester, K.A. et al. Phage libraries for generation of clinically useful antibodies. Lancet 343, 455–456 (1994).

  23. 23

    Kettleborough, C.A., Saldanha, J., Heath, V.J., Morrison, C.J. & Bendig, M.M. Humanization of a mouse monoclonal antibody by CDR-grafting: the importance of framework residues on loop conformation. Protein Eng. 4, 773–783 (1991).

  24. 24

    Wang, G. et al. A T cell-independent antitumor response in mice with bone marrow cells retrovirally transduced with an antibody/Fc-gamma chain chimeric receptor gene recognizing a human ovarian cancer antigen. Nat. Med. 4, 168–172 (1998).

  25. 25

    Wang, X. et al. Cell fusion is the principal source of bone-marrow-derived hepatocytes. Nature 422, 897–901 (2003).

  26. 26

    Mehta, K., Shahid, U. & Malavasi, F. Human CD38, a cell-surface protein with multiple functions. FASEB J. 10, 1408–1417 (1996).

  27. 27

    Obrink, B. CEA adhesion molecules: multifunctional proteins with signal-regulatory properties. Curr. Opin. Cell Biol. 9, 616–626 (1997).

  28. 28

    Carpenter, G. Receptor tyrosine kinase substrates: src homology domains and signal transduction. FASEB J. 6, 3283–3289 (1992).

  29. 29

    Kemper, C. et al. Activation of human CD4+ cells with CD3 and CD46 induces a T-regulatory cell 1 phenotype. Nature 421, 388–392 (2003).

  30. 30

    Schneider, U., Bullough, F., Vongpunsawad, S., Russell, S.J. & Cattaneo, R. Recombinant measles viruses efficiently entering cells through targeted receptors. J. Virol. 74, 9928–9936 (2000).

  31. 31

    Robbins, P.F. et al. Transduction and expression of the human carcinoembryonic antigen gene in a murine colon carcinoma cell line. Cancer Res. 51, 3657–3662 (1991).

  32. 32

    Cathomen, T., Buchholz, C.J., Spielhofer, P. & Cattaneo, R. Preferential initiation at the second AUG of the measles virus F mRNA: a role for the long untranslated region. Virology 214, 628–632 (1995).

  33. 33

    Mizuguchi, H., Xu, Z.L., Sakurai, F., Mayumi, T. & Hayakawa, T. Tight positive regulation of transgene expression by a single adenovirus vector containing the rtTA and tTS expression cassettes in separate genome regions. Hum. Gene Ther. 14, 1265–1277 (2003).

  34. 34

    Firsching, R. et al. Measles virus spread by cell-cell contacts: uncoupling of contact-mediated receptor (CD46) downregulation from virus uptake. J Virol. 73, 5265–5273 (1999).

  35. 35

    Kanegae, Y. et al. Efficient gene activation in mammalian cells by using recombinant adenovirus expressing site-specific Cre recombinase. Nucl. Acids Res. 23, 3816–3821 (1995).

  36. 36

    Mittereder, N., March, K.L. & Trapnell, B.C. Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J. Virol. 70, 7498–7509 (1996).

  37. 37

    Langedijk, J.P., Daus, F.J. & van Oirschot, J.T. Sequence and structure alignment of Paramyxoviridae attachment proteins and discovery of enzymatic activity for a morbillivirus hemagglutinin. J. Virol. 71, 6155–6167 (1997).

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We thank C.D. James for CHO-EGFR cells, Y. Yanagi for CHO-SLAM cells, J. Schlom for MC38-CEA cells, E. Vitetta for SKOV3ip.1 cells, J.P. Atkinson for CD46 plasmid, J.A. Lust for CD38 scFv, R. Hawkins for CEA scFv and G. Winter for EGFR scFv. We also thank M.J. Federspiel and R.G. Vile for critical reading of the manuscript. This study is supported by the Mayo Foundation, Harold W. Siebens Foundation and NIH grants CA100634-01 and CA90636-01.

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Correspondence to Stephen J Russell.

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Nakamura, T., Peng, K., Vongpunsawad, S. et al. Antibody-targeted cell fusion. Nat Biotechnol 22, 331–336 (2004) doi:10.1038/nbt942

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