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Journal of Virology, August 2002, p. 7506-7517, Vol. 76, No. 15
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.15.7506-7517.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Robust Recall and Long-Term Memory T-Cell Responses Induced by Prime-Boost Regimens with Heterologous Live Viral Vectors Expressing Human Immunodeficiency Virus Type 1 Gag and Env Proteins

Karl Haglund,1,2 Ingrid Leiner,3,4,{dagger} Kristen Kerksiek,4,{ddagger} Linda Buonocore,1 Eric Pamer,3,4,{dagger} and John K. Rose1,5*

Departments of Pathology,1 Cell Biology,5 Medicine,3 Program in Microbiology,2 Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 065104

Received 29 January 2002/ Accepted 24 April 2002

We investigated long-term memory and recall cellular immune responses to human immunodeficiency virus type 1 (HIV-1) Env and Gag proteins elicited by recombinant vesicular stomatitis viruses (VSVs) expressing Env and Gag. More than 7 months after a single vaccination with VSV-Env, ~6% of CD8+ splenocytes stained with major histocompatibility complex class I tetramers containing the Env p18-I10 immunodominant peptide and showed a memory phenotype (CD44Hi). The level of tetramer-positive cells in memory was about 14% of the peak primary response. Recall responses elicited in these mice 5 days after boosting with a heterologous recombinant vaccinia virus expressing HIV-1 Env showed that 40 to 45% of CD8+ splenocytes were tetramer positive and activated (CD62LLo), and these cells produced gamma interferon after stimulation with Env peptide, indicating that they were functional. Five months after the boost, the long-term memory cell population (tetramer positive, CD44Hi) constituted 30% of the CD8+ splenocytes. Recall responses to HIV-1 Gag were examined in mice primed with VSV recombinants expressing HIV-1 Gag protein and boosted with a vaccinia virus recombinant expressing Gag. Using this protocol, we found that ~40% of CD8+ splenocytes were activated (CD62LLo) and specific for a Gag immunodominant peptide (tetramer positive). The high-level Gag recall response elicited by the vaccinia virus-Gag was greater than that obtained by boosting with a VSV-Gag vector with a different VSV glycoprotein. The corresponding levels of CD44Hi memory cells were also higher long after boosting with vaccinia virus-Gag than after boosting with a glycoprotein exchange VSV-Gag. Our results show that VSV vectors elicit high-level memory CTL responses and that these can be amplified as much as six- to sevenfold using a heterologous boosting vector.

* Corresponding author. Mailing address: Departments of Pathology and Cell Biology, Yale University School of Medicine, New Haven, CT 06510. Phone: (203) 785-6794. Fax: (203) 785-7467. E-mail: jrose{at}biomed.med.yale.edu.

{dagger} Present address: Infectious Diseases Service, Laboratory of Antimicrobial Immunity, Memorial Sloan-Kettering Cancer Center, New York, NY 10021.

{ddagger} Present address: LMU München, Institut für Immunologie, 803360 Munich, Germany.

Journal of Virology, August 2002, p. 7506-7517, Vol. 76, No. 15
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.15.7506-7517.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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