A sensitive and versatile bioluminescence bioassay for HIV type 1 based on adenoviral vectors.
The construction and characterization of a versatile bioassay for the quantification of HIV-1 viral infection and HIV-1 Tat protein activity based on recombinant adenoviral vectors carrying an HIV LTR-driven luciferase reporter gene is described. The assay system consists of a set of two adeno-reporter vectors, one of which is responsive to HIV-1 Tat protein activity, and the second of which is not, by virtue of a deletion of the TAR site within the HIV LTR. This configuration of the reporter genes allows one to distinguish Tat-specific activation from Tat-non-specific HIV LTR-mediated gene expression. The adenoviral HIV LTR-mediated luciferase gene expression is highly responsive to Tat and increases linearly with increasing levels of HIV-1 infection, reaching levels of between 3- and 1000-fold induction. The adeno-reporter viruses can be utilized to detect Tat activity and HIV-1 infection in a wide range of cell types, including 293, CEM, HUT-78, Jurkat, and HeLa-derived cell lines. The resulting bioassay is convenient, sensitive, and readily adaptable to automated procedures. These characteristics of the adeno-reporter assay make it a valuable reagent for studies of HIV infection and for analysis of HIV-inhibitory agents.
Axelrod
J H
JH
Department of Virology, The Hebrew University-Hadassah Medical School, Jerusalem, Israel. Axelrod@hadassah.org.il
Honigman
A
A
D000256:Adenoviridae / Q000235:genetics*; D019380:Anti-HIV Agents / Q000494:pharmacology; D001681:Biological Assay; D002460:Cell Line; D015696:Gene Products, tat / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D005822:Genetic Vectors; D015658:HIV Infections / Q000175:diagnosis* / Q000821:virology; D016325:HIV Long Terminal Repeat / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements; D015215:Zidovudine / Q000494:pharmacology; D054322:tat Gene Products, Human Immunodeficiency Virus
D019380:Anti-HIV Agents; D015696:Gene Products, tat; D054322:tat Gene Products, Human Immunodeficiency Virus; D015215:Zidovudine; D008156:Luciferases