A luciferase-reporter gene-expressing T-cell line facilitates neutralization and drug-sensitivity assays that use either R5 or X4 strains of human immunodeficiency virus type 1.
We describe the production and properties of a permanent cell line, CEM.NKR-CCR5-Luc. This line is a derivative of the CEM.NKR-CCR5 line, stably transfected to express the luciferase reporter gene under the transcriptional control of the HIV-2 LTR. Thus the cells respond to Tat expression during HIV-1 infection by producing luciferase, a protein that can be readily and accurately quantitated in a luminometer. The CEM.NKR-CCR5-Luc line expresses both the CCR5 and CXCR4 coreceptors and can therefore be infected with a range of HIV-1 isolates, irrespective of their tropism properties. This is true of HIV-1 isolates from several genetic subtypes and also of a group O isolate. Furthermore, luciferase expression is also activated by infection of the cells with SIVmac239 or SIVmac251. We show that the CEM.NKR-CCR5-Luc cells can be used in assays of HIV-1 neutralization and also for identifying inhibitors of HIV-1 entry targeted at the CCR5 and CXCR4 coreceptors. The luciferase end point simplifies the performance of neutralization and inhibitor-screening assays compared to the use of more conventional end points such as the detection of extracellular p24 antigen.
Spenlehauer
C
C
Weill Medical College of Cornell University, New York, New York, USA.
Gordon
C A
CA
Trkola
A
A
Moore
J P
JP
D000577:Amides / Q000494:pharmacology; D019380:Anti-HIV Agents / Q000494:pharmacology*; D000914:Antibodies, Viral / Q000276:immunology*; D001596:Benzylamines; D065100:CCR5 Receptor Antagonists; D002460:Cell Line; D002461:Cell Line, Transformed; D000080027:Cyclams; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015870:Gene Expression; D017930:Genes, Reporter; D016655:HIV Core Protein p24 / Q000378:metabolism; D015497:HIV-1 / Q000187:drug effects* / Q000276:immunology* / Q000302:isolation & purification / Q000502:physiology; D006571:Heterocyclic Compounds / Q000494:pharmacology; D006801:Humans; D008156:Luciferases / Q000235:genetics*; D009500:Neutralization Tests; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D019713:Receptors, CCR5 / Q000235:genetics / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors*; D013601:T-Lymphocytes / Q000166:cytology / Q000821:virology*
D000577:Amides; D019380:Anti-HIV Agents; D000914:Antibodies, Viral; D001596:Benzylamines; D065100:CCR5 Receptor Antagonists; D000080027:Cyclams; D016655:HIV Core Protein p24; D006571:Heterocyclic Compounds; D000644:Quaternary Ammonium Compounds; D019713:Receptors, CCR5; D019718:Receptors, CXCR4; C119369:TAK 779; D008156:Luciferases; C088327:plerixafor