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Clinical and Vaccine Immunology, August 2006, p. 845-853, Vol. 13, No. 8
1071-412X/06/$08.00+0     doi:10.1128/CVI.00072-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Plant-Produced Cottontail Rabbit Papillomavirus L1 Protein Protects against Tumor Challenge: a Proof-of-Concept Study

T. Kohl,1,2 I. I. Hitzeroth,1,2 D. Stewart,2 A. Varsani,3 V. A. Govan,2 N. D. Christensen,4 A.-L. Williamson,2,5 and E. P. Rybicki1,2*

Department of Molecular and Cell Biology, University of Cape Town, Rondebosch 7701, South Africa,1 Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Observatory 7925, Cape Town, South Africa,2 Electron Microscopy Unit, University of Cape Town, Rondebosch 7701, South Africa,3 Department of Pathology, The Jake Gittlen Cancer Research Institute, and Department of Microbiology and Immunology, College of Medicine, Pennsylvania State University, Hershey, Pennsylvania 17033,4 National Health Laboratory Services, Groote Schuur Hospital, Observatory 7925, Cape Town, South Africa5

Received 24 February 2006/ Returned for modification 7 April 2006/ Accepted 9 June 2006

The native cottontail rabbit papillomavirus (CRPV) L1 capsid protein gene was expressed transgenically via Agrobacterium tumefaciens transformation and transiently via a tobacco mosaic virus (TMV) vector in Nicotiana spp. L1 protein was detected in concentrated plant extracts at concentrations up to 1.0 mg/kg in transgenic plants and up to 0.4 mg/kg in TMV-infected plants. The protein did not detectably assemble into viruslike particles; however, immunoelectron microscopy showed presumptive pentamer aggregates, and extracted protein reacted with conformation-specific and neutralizing monoclonal antibodies. Rabbits were injected with concentrated protein extract with Freund's incomplete adjuvant. All sera reacted with baculovirus-produced CRPV L1; however, they did not detectably neutralize infectivity in an in vitro assay. Vaccinated rabbits were, however, protected against wart development on subsequent challenge with live virus. This is the first evidence that a plant-derived papillomavirus vaccine is protective in an animal model and is a proof of concept for human papillomavirus vaccines produced in plants.

* Corresponding author. Mailing address: Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, Cape Town 7701, South Africa. Phone: 27 (0) 21 650 3265. Fax: 27 (0) 21 650 2646. E-mail: ed{at}science.uct.ac.za.

Clinical and Vaccine Immunology, August 2006, p. 845-853, Vol. 13, No. 8
1071-412X/06/$08.00+0     doi:10.1128/CVI.00072-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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