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Internalization of a Bacillus anthracis Protective Antigen-c-Myc Fusion Protein Mediated by Cell Surface Anti-c-Myc Antibodies

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Abstract

Background

Anthrax toxin, secreted by Bacillus anthracis, consists of protective antigen (PA) and either lethal factor (LF) or edema factor (EF). PA, the receptor-binding component of the toxin, translocates LF or EF into the cytosol, where the latter proteins exert their toxic effects. We hypothesized that anthrax toxin fusion proteins could be used to kill virus-infected cells and tumor cells, if PA could be redirected to unique receptors found only on these cells.

Materials And Methods

To test this hypothesis in a model system, amino acids 410–419 of the human p62c-myc epitope were fused to the C-terminus of PA to redirect PA to the c-Myc-specific hybridoma cell line 9E10.

Results

The PA-c-Myc fusion protein killed both mouse macrophages and 9E10 hybridoma cells when administered with LF or an LF fusion protein (FP59), respectively. Similar results were obtained with PA, which suggests that PA-c-Myc used the endogenous PA receptor to enter the cells. By blocking the endogenous PA receptors on 9E10 cells with the competitive inhibitor PA SNKEΔFF, the PA-c-Myc was directed to an alternate receptor, i.e., the anti-c-Myc antibodies presented on the cell surface. The c-Myc IgG were proven to act as receptors because the addition of a synthetic peptide containing the c-Myc epitope along with PA SNKEΔFF further reduced the toxicity of PA-c-Myc + FP59.

Conclusion

This study shows that PA can be redirected to alternate receptors by adding novel epitopes to the C-terminus of PA, enabling the creation of cell-directed toxins for therapeutic purposes.

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Acknowledgments

The authors are grateful to Valery Gordon for helpful discussions in the preparation of this manuscript.

Author information

Author notes

  1. Angela Chi

    Present address: Harvard School of Dental Medicine, 188 Longwood Avenue, Boston, MA, 02115, USA

  2. Avelino V. Teixeira

    Present address: Division of Nephrology, Mount Sinai School of Medicine, One Gustave L. Levy Place, Box 1243, New York, NY, 10029-6574, USA

  3. Peter J. Nicholls

    Present address: Department of Biosciences, University of Kent at Canterbury, Canterbury, Kent, CT2 7NJ, UK

Authors and Affiliations

  1. Oral Infection and Immunity Branch, National Institute of Dental Research, NIH, Bethesda, Maryland, USA

    Mini Varughese, Angela Chi, Avelino V. Teixeira, Peter J. Nicholls, Jerry M. Keith & Stephen H. Leppla

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  1. Mini Varughese
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Correspondence to Stephen H. Leppla.

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Communicated by I. Pasten.

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Varughese, M., Chi, A., Teixeira, A.V. et al. Internalization of a Bacillus anthracis Protective Antigen-c-Myc Fusion Protein Mediated by Cell Surface Anti-c-Myc Antibodies. Mol Med 4, 87–95 (1998). https://doi.org/10.1007/BF03401732

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