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Journal of Virology, August 2005, p. 10108-10125, Vol. 79, No. 16
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.16.10108-10125.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Human Immunodeficiency Virus Type 1 env Clones from Acute and Early Subtype B Infections for Standardized Assessments of Vaccine-Elicited Neutralizing Antibodies

Ming Li,1 Feng Gao,2 John R. Mascola,3 Leonidas Stamatatos,4 Victoria R. Polonis,5 Marguerite Koutsoukos,6 Gerald Voss,6 Paul Goepfert,7 Peter Gilbert,8 Kelli M. Greene,1 Miroslawa Bilska,1 Denise L. Kothe,7 Jesus F. Salazar-Gonzalez,7 Xiping Wei,7 Julie M. Decker,7 Beatrice H. Hahn,7 and David C. Montefiori1*

Departments of Surgery,1 Medicine, Duke University Medical Center, Durham, North Carolina 27710,2 Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892,3 Seattle Biomedical Research Institute, Seattle, Washington 98109,4 Henry M. Jackson Foundation, Rockville, Maryland 20850,5 GlaxoSmithKline Biologicals, 1330 Rixensart, Belgium,6 Department of Medicine, University of Alabama, Birmingham, Alabama 35294,7 Statistical Center for HIV/AIDS Research and Prevention, University of Washington, Seattle, Washington 981098

Received 22 February 2005/ Accepted 9 May 2005

Induction of broadly cross-reactive neutralizing antibodies is a high priority for AIDS vaccine development but one that has proven difficult to be achieved. While most immunogens generate antibodies that neutralize a subset of T-cell-line-adapted strains of human immunodeficiency virus type 1 (HIV-1), none so far have generated a potent, broadly cross-reactive response against primary isolates of the virus. Even small increments in immunogen improvement leading to increases in neutralizing antibody titers and cross-neutralizing activity would accelerate vaccine development; however, a lack of uniformity in target strains used by different investigators to assess cross-neutralization has made the comparison of vaccine-induced antibody responses difficult. Thus, there is an urgent need to establish standard panels of HIV-1 reference strains for wide distribution. To facilitate this, full-length gp160 genes were cloned from acute and early subtype B infections and characterized for use as reference reagents to assess neutralizing antibodies against clade B HIV-1. Individual gp160 clones were screened for infectivity as Env-pseudotyped viruses in a luciferase reporter gene assay in JC53-BL (TZM-bl) cells. Functional env clones were sequenced and their neutralization phenotypes characterized by using soluble CD4, monoclonal antibodies, and serum samples from infected individuals and noninfected recipients of a recombinant gp120 vaccine. Env clones from 12 R5 primary HIV-1 isolates were selected that were not unusually sensitive or resistant to neutralization and comprised a wide spectrum of genetic, antigenic, and geographic diversity. These reference reagents will facilitate proficiency testing and other validation efforts aimed at improving assay performance across laboratories and can be used for standardized assessments of vaccine-elicited neutralizing antibodies.

* Corresponding author. Mailing address: Department of Surgery, Laboratory for AIDS Vaccine Research and Development, P.O. Box 2926, Duke University Medical Center, Durham, NC 27710. Phone: (919) 684-5278. Fax: (919) 684-4288. E-mail: monte{at}acpub.duke.edu.

Journal of Virology, August 2005, p. 10108-10125, Vol. 79, No. 16
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.16.10108-10125.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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