Characterization of a chemokine receptor CCR5‐negative T cell line and its use in determining human immunodeficiency virus type 1 phenotype

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Abstract
A human CD4‐positive T cell line from a donor homozygous negative for the chemokine receptor CCR5 was established, characterized, and used for determining the coreceptor usage of human immunodeficiency virus type 1 (HIV‐1) isolates. Clones of this IL‐2 dependent human T‐cell lymphotropic virus type 1 (HTLV‐I) immortalized cell line, named IsnoR5 clones 1 and 2, are susceptible to infection by HIV‐1 isolates that use CXCR4 as a coreceptor but resistant to infection by CCR5 tropic HIV‐1 viruses. HIV‐1 isolates whose replication is inhibited in IsnoR5 cells in the presence of the bicyclam AMD 3100, a CXCR4 specific inhibitor, utilize a coreceptor distinct from CCR5 and CXCR4. Using a panel of primary HIV‐1 isolates we have shown that a single T cell line is sufficient to discriminate between use of CCR5, CXCR4 or an alternative coreceptor. As IsnoR5 clone 1 cells revealed the existence of even minor populations of CXCR4‐using virus variants, they could be useful for the early identification of changes in coreceptor usage in HIV infected individuals facilitating the timely introduction of appropriate clinical treatments. J. Med. Virol. 80:192–200, 2008.

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