TY - JOUR
T1 - LuSIV cells
T2 - A reporter cell line for the detection and quantitation of a single cycle of HIV and SIV replication
AU - Roos, J. W.
AU - Maughan, M. F.
AU - Liao, Z.
AU - Hildreth, J. E.K.
AU - Clements, J. E.
N1 - Funding Information:
The authors thank Dr. Barry Margulies for his invaluable insight in designing the LuSIV assay system, Dr. Deborah McClellan for her editorial support, and Maryann Brooks for her help in preparing the manuscript for publication. This work was supported by an NIH training Grant NSO7392 and Grants NS35751, NS38008, and HL061962.
PY - 2000/8/1
Y1 - 2000/8/1
N2 - A single cycle of viral replication is the time required for a virus to enter the host cell, replicate its genome, and produce infectious progeny virions. The primate lentiviruses, human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV), require on average 24 h to complete one cycle of replication. We have now developed and characterized a reporter assay system in CEMx174 cells for the quantitative measurement of HIV/SIV infection within a single replication cycle. The SIV(mac)239 LTR (-225 → + 149) was cloned upstream of the firefly luciferase reporter gene and this reporter plasmid is maintained in CEMx174 cells under stable selection. This cell line, designated LuSIV, is highly sensitive to infection by primary and laboratory strains of HIV/SIV, resulting in Tat-mediated expression of luciferase, which correlates with viral infectivity. Furthermore, manipulation of LuSIV ceils for the detection of luciferase activity is easy to perform and requires a minimal amount of time as compared to current HIV/SIV detection systems. The LuSIV system is a powerful tool for the analysis of HIV/SIV infection that provides a unique assay system that can detect virus replication prior to 24 h and does not require virus to spread from cell to cell. Thus these ceils can be used for the study of replication-deficient viruses and the high throughput screening of antivirals, or other inhibitors of infection. (C) 2000 Academic Press.
AB - A single cycle of viral replication is the time required for a virus to enter the host cell, replicate its genome, and produce infectious progeny virions. The primate lentiviruses, human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV), require on average 24 h to complete one cycle of replication. We have now developed and characterized a reporter assay system in CEMx174 cells for the quantitative measurement of HIV/SIV infection within a single replication cycle. The SIV(mac)239 LTR (-225 → + 149) was cloned upstream of the firefly luciferase reporter gene and this reporter plasmid is maintained in CEMx174 cells under stable selection. This cell line, designated LuSIV, is highly sensitive to infection by primary and laboratory strains of HIV/SIV, resulting in Tat-mediated expression of luciferase, which correlates with viral infectivity. Furthermore, manipulation of LuSIV ceils for the detection of luciferase activity is easy to perform and requires a minimal amount of time as compared to current HIV/SIV detection systems. The LuSIV system is a powerful tool for the analysis of HIV/SIV infection that provides a unique assay system that can detect virus replication prior to 24 h and does not require virus to spread from cell to cell. Thus these ceils can be used for the study of replication-deficient viruses and the high throughput screening of antivirals, or other inhibitors of infection. (C) 2000 Academic Press.
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U2 - 10.1006/viro.2000.0431
DO - 10.1006/viro.2000.0431
M3 - Article
C2 - 10915601
AN - SCOPUS:0034255169
SN - 0042-6822
VL - 273
SP - 307
EP - 315
JO - Virology
JF - Virology
IS - 2
ER -