Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Reexamination Certificate
2007-07-31
2007-07-31
Chen, Stacy B. (Department: 1648)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
C424S208100
Reexamination Certificate
active
10656803
ABSTRACT:
The present invention features methods and compositions relating to a virion-based fusion assay for detection of infection of a target cell by an enveloped retroviral virion such as HIV. The assay uses virions containing a chimeric viral protein comprising a viral accessory polypeptide (such as Vpr) fused to a reporter polypeptide (such as beta-lactamase). Fusion of the virion with a target cell membrane results in intracellular delivery of the chimeric protein to the target cell, which in turn provides for detection of a detectable signal mediated by the reporter polypeptide portion of the chimeric polypeptide. Significant detectable signal is only detected following intracellular delivery of the chimeric viral protein, thus providing for detection of productive viral entry to the exclusion of non-productive, endocytic entry of virions into the cell.
REFERENCES:
Zlokarnik et al., Quantitation of transcription and clonal selection of single living cells with beta-lactamase as reporter. 1998, vol. 279, p. 84-88.
Chen, Y.D., et al., “On the Use of Self-Quenching Fluorophores in the Study of Membrane Fusion Kinetics—The Effect of Slow Probe Redistribution,” Biophys. Chem. 34:283-292 (1989).
Fackler, O. T. et al., “Endocytic Entry of HIV-1,” Current Biol. 10:1005-1008 (2000).
Klatzmann, D. R., et al., “The CD4 Molecule and HIV Infection,” Immunodef., Rev. 2, 43-66 (1990).
Kowalski, M., et al., “Functional Regions of the Envelope Glycoprotein of Human Immunodeficiency Virus Type 1,” Science 237:1351 (1987).
Lifson, J. D., et al., “Induction of CD4-Dependent Cell Fusion by the HTLV-III/LAV Envelope Glycoprotein,” Nature 323:725 (1986).
Lowy, R.J., et al., “Observation of Single Influenza Virus-Cell Fusion and Measurement by Fluorescence Video Microscopy,” PNAS 87:1850-1854 (1990).
Maddon, P. J., et al., “The T4 Gene Encodes the AIDS Virus Receptor and is Expressed in the Immune System and the Brain,” Cell 47, 333-348 (1986).
Maddon, P. J., et al., “The Isolation and Nucleotide Sequence of a cDNA Encoding the T Cell Surface Protein T4: A New Member of the Immunoglobulin Gene Family,” Cell 42, 93-104 (1985).
Maddon, P. J., et al., “HIV Infection Does Not Require Endocytosis of its Receptor, CD4,” Cell 54:865 (1988).
Marchal, V., et al., “Human Immunodeficiency Virus Type 1 Entry into Macrophages Mediated by Macropinocytosis,” J. Virology 75:11166-11177 (2001).
Muthumani et al. “Vpr-GFP Virion Particle Identifies HIV-Infected Targets and Preserves HIV-IV pr Function in Macrophages and T-Cells,” DNA Cell Biol. 19:179-188 (2000).
Raviv, Y., et al., “Quantitative Measurement of Fusion of HIV-1 and SIV with Cultured Cells Using Photosensitized Labeling,” Virology 293:243-251 (2002).
Wain-Hobson, S., et al., “Nucleotide Sequence of the AIDS Virus, LAV,” Cell 40, 9-17 (1985).
Cavrois Marielle
de Noronha Carlos
Greene Warner C.
Boesen Agnieszka
Bozicevic Field & Francis LLP
Chen Stacy B.
Francis Carol L.
Schwartz Richard A.
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