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Journal of Virology, December 2003, p. 12782-12794, Vol. 77, No. 23
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.23.12782-12794.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Covalently Linked Human Immunodeficiency Virus Type 1 gp120/gp41 Is Stably Anchored in Rhabdovirus Particles and Exposes Critical Neutralizing Epitopes

Philip M. McKenna,1,2 Roger J. Pomerantz,2,3,4 Bernhard Dietzschold,1,2 James P. McGettigan,2,3 and Matthias J. Schnell2,3*

Departments of Microbiology,1 Biochemistry and Molecular Pharmacology,3 Medicine,4 Center for Human Virology and Biodefense, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 191072

Received 22 May 2003/ Accepted 22 August 2003

Rabies virus (RV) vaccine strain-based vectors show significant promise as potential live-attenuated vaccines against human immunodeficiency virus type 1 (HIV-1). Here we describe a new RV construct that will also likely have applications as a live-attenuated or killed-particle immunogen. We have created a RV containing a chimeric HIV-1 Env protein, which contains introduced cysteine residues that give rise to an intermolecular disulfide bridge between gp120 and the ectodomain of gp41. This covalently linked gp140 (gp140 SOS) is fused in frame to the cytoplasmic domain of RV G glycoprotein and is efficiently incorporated into the RV virion. On the HIV-1 virion, the gp120 and gp41 moieties are noncovalently associated, which leads to extensive shedding of gp120 from virions and virus-infected cells. The ability to use HIV-1 particles as purified, inactivated immunogens has been confounded by the loss of gp120 during preparation. Additionally, monomeric gp120 and uncleaved gp160 molecules have been shown to be poor antigenic representations of virion-associated gp160. Because the gp120 and gp41 portions are covalently attached in the gp140 SOS molecule, the protein is maintained on the surface of the RV virion throughout purification. Surface immunostaining and fluorescence-activated cell sorting analysis with anti-envelope antibodies show that the gp140 SOS protein is stably expressed on the surface of infected cells and maintains CD4 binding capabilities. Furthermore, Western blot and immunoprecipitation experiments with infected-cell lysates and purified virions show that a panel of neutralizing anti-envelope antibodies efficiently recognize the gp140 SOS protein. The antigenic properties of this recombinant RV particle containing covalently attached Env, as well as the ability to present Env in a membrane-bound form, suggest that this approach could be a useful component of a HIV-1 vaccine strategy.

* Corresponding author. Mailing address: 350 BLSB, 233 South 10th St., Philadelphia, PA 19107. Phone: (215) 503-1260. Fax: (215) 923-1956. E-mail: matthias.schnell{at}jefferson.edu.

Journal of Virology, December 2003, p. 12782-12794, Vol. 77, No. 23
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.23.12782-12794.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Faber, M., Lamirande, E. W., Roberts, A., Rice, A. B., Koprowski, H., Dietzschold, B., Schnell, M. J. (2005). A single immunization with a rhabdovirus-based vector expressing severe acute respiratory syndrome coronavirus (SARS-CoV) S protein results in the production of high levels of SARS-CoV-neutralizing antibodies. J. Gen. Virol. 86: 1435-1440 [Abstract] [Full Text]  
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