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Journal of Virology, June 2005, p. 7121-7134, Vol. 79, No. 11
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.11.7121-7134.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Differences in the Fitness of Two Diverse Wild-Type Human Immunodeficiency Virus Type 1 Isolates Are Related to the Efficiency of Cell Binding and Entry

Andre J. Marozsan,1,2,{dagger} Dawn M. Moore,1,3,{dagger} Michael A. Lobritz,1,3 Erika Fraundorf,1 Awet Abraha,1 Jacqueline D. Reeves,4 and Eric J. Arts1,2,3*

Division of Infectious Diseases, Department of Medicine,1 Department of Pharmacology,2 Molecular and Cellular Biology Training Program, Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, Ohio,3 Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania4

Received 2 October 2004/ Accepted 18 January 2005

The ability of one primary human immunodeficiency virus type 1 (HIV-1) isolate to outcompete another in primary CD4+ human lymphoid cells appears to be mediated by the efficiency of host cell entry. This study was designed to test the role of entry on fitness of wild-type HIV-1 isolates (e.g., replicative capacity) and to examine the mechanism(s) involved in differential entry efficiency. The gp120 coding regions of two diverse HIV-1 isolates (the more-fit subtype B strain, B5-91US056, and less-fit C strain, C5-97ZA003) were cloned into a neutral HIV-1 backbone by using a recently described yeast cloning technique. The fitness of the primary B5 HIV-1 isolates and its env gene cloned into the NL4-3 laboratory strain had similar fitness, and both were more fit than the C5 primary isolate and its env/NL4-3 chimeric counterpart. Increased fitness of the B5 over C5 virus was mediated by the gp120 coding region of the env gene. An increase in binding/fusion, as well as decreased sensitivity to entry inhibitors (PSC-RANTES and T-20), was observed in cell fusion assays mediated by B5 gp120 compared to C5 gp120. Competitive binding assays using a novel whole virus-cell system indicate that the primary or chimeric B5 had a higher avidity for CD4/CCR5 on host cells than the C5 counterpart. This increased avidity of an HIV-1 isolate for its cell receptors may be a significant factor influencing overall replicative capacity or fitness.

* Corresponding author. Mailing address: Division of Infectious Diseases, BRB 1029, Case Western Reserve University, 10900 Euclid Ave., Cleveland, OH 44106. Phone: (216) 368-8904. Fax: (216) 368-2034. E-mail: eja3{at}po.cwru.edu.

{dagger} A.J.M. and D.M.M. contributed equally to this study.

Journal of Virology, June 2005, p. 7121-7134, Vol. 79, No. 11
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.11.7121-7134.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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