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Journal of Virology, November 1999, p. 8966-8974, Vol. 73, No. 11
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

A Cell Line-Based Neutralization Assay for Primary Human Immunodeficiency Virus Type 1 Isolates That Use either the CCR5 or the CXCR4 Coreceptor

Alexandra Trkola,1,2 Jamie Matthews,1 Cynthia Gordon,1,3 Tom Ketas,1 and John P. Moore1,2,*

The Aaron Diamond AIDS Research Center,1 The Rockefeller University,2 and Department of Pathology, New York University School of Medicine,3 New York, New York

Received 14 June 1999/Accepted 6 August 1999

We describe here a cell line-based assay for the evaluation of human immunodeficiency virus type 1 (HIV-1) neutralization. The assay is based on CEM.NKR cells, transfected to express the HIV-1 coreceptor CCR5 to supplement the endogenous expression of CD4 and the CXCR4 coreceptor. The resulting CEM.NKR-CCR5 cells efficiently replicate primary HIV-1 isolates of both R5 and X4 phenotypes. A comparison of the CEM.NKR-CCR5 cells with mitogen-activated peripheral blood mononuclear cells (PBMC) in neutralization assays with sera from HIV-1-infected individuals or specific anti-HIV-1 monoclonal antibodies shows that the sensitivity of HIV-1 neutralization is similar in the two cell types. The CEM.NKR-CCR5 cell assay, however, is more convenient to perform and eliminates the donor-to-donor variation in HIV-1 replication efficiency, which is one of the principal drawbacks of the PBMC-based neutralization assay. We suggest that this new assay is suitable for the general measurement of HIV-1 neutralization by antibodies.

* Corresponding author. Mailing address: The Aaron Diamond AIDS Research Center, 455 First Ave., New York, NY 10021. Phone: (212) 725-0018. Fax: (212) 725-1126. E-mail: jmoore{at}adarc.org.

Journal of Virology, November 1999, p. 8966-8974, Vol. 73, No. 11
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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