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Journal of Virology, March 2003, p. 3157-3166, Vol. 77, No. 5
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.5.3157-3166.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Tat-Neutralizing Antibodies in Vaccinated Macaques

Ilia Tikhonov, Tracy J. Ruckwardt, Glen S. Hatfield, and C. David Pauza*

Institute of Human Virology, University of Maryland Biotechnology Institute, Baltimore, Maryland 21201

Received 12 August 2002/ Accepted 20 November 2002

The human immunodeficiency virus Tat protein is essential for virus replication and is a candidate vaccine antigen. Macaques immunized with Tat or chemically modified Tat toxoid having the same clade B sequence developed strong antibody responses. We compared these antisera for their abilities to recognize diverse Tat sequences. An overlapping peptide array covering three clade B and two clade C Tat sequences was constructed to help identify reactive linear epitopes. Sera from Tat-immunized macaques were broadly cross-reactive with clade B and clade C sequences but recognized a clade B-specific epitope in the basic domain. Sera from Tat toxoid-immunized macaques had a more restricted pattern of recognition, reacting mainly with clade B and with only one clade B basic domain sequence, which included the rare amino acids RPPQ at positions 57 to 60. Monoclonal antibodies against the amino terminus or the domain RPPQ sequence blocked Tat uptake into T cells and neutralized Tat in a cell-based transactivation assay. Macaques immunized with Tat or Tat toxoid proteins varied in their responses to minor epitopes, but all developed a strong response to the amino terminus, and antisera were capable of neutralizing Tat in a transactivation assay.

* Corresponding author. Mailing address: Institute of Human Virology, University of Maryland Biotechnology Institute, 725 W. Lombard St., Rm. N546, Baltimore, MD 21201. Phone: (410) 706-1367. Fax: (410) 707-6212. E-mail: pauza{at}umbi.umd.edu.

Journal of Virology, March 2003, p. 3157-3166, Vol. 77, No. 5
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.5.3157-3166.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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