Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Animal cell – per se – expressing immunoglobulin – antibody – or...
Reexamination Certificate
2001-02-07
2002-09-17
Park, Hankyel T. (Department: 1648)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Animal cell, per se, expressing immunoglobulin, antibody, or...
C435S006120, C435S039000, C424S009200
Reexamination Certificate
active
06451598
ABSTRACT:
TECHNICAL FIELD
This invention relates generally to the field of drug screening assays, more particularly to antiviral drug screening assays and specifically to cell fusion based antiviral drug screening assays.
BACKGROUND OF THE INVENTION
Cell fusion assays designed to identify viral inhibitory agents, i.e., agents that inhibit the binding of a viral glycoprotein and a target cell receptor as well as the virion-cell fusion that follows this binding event, are known in the art. In cell fusion assays, a first cell expresses a viral envelope coat protein on its surface and represents the “virion” while a second cell expresses a receptor for the viral coat protein on its surface and represents the target cell. When brought together under appropriate conditions, the first and second cells fuse via a viral coat protein/target cell receptor mediated event.
Many of the cell fusion assays currently employed are “vaccinia” based fusion assays in which the envelope protein of the virion of interest is only transiently expressed in the virion host cell. While such vaccinia based cell fusion assays have been extremely useful, they do suffer from certain disadvantages, including an unsuitability for adaptation to high throughput automated formats.
As such, there is continued interest in the identification of additional cell fusion assay protocols. Of particular interest would be the development of protocol that are adaptable to high throughput, automated formats.
Relevant Literature
Papers of interest include: Berger et al., HIV: A Practical Approach (J. Karn ed. IRL Press) Vol. 2:123-145 (1995); Blondell et al., Abstract from
3
rd
Annual Conference on AIDS research in California, Feb. 25, 2000; Weiss et al., J. Virol. (1993) 67: 7060-7066; and Weiss et al., AIDS (1996) 10:241-246.
SUMMARY OF THE INVENTION
Cell fusion assays for identifying antiviral compounds, particularly enveloped virus inhibitory compounds, are provided. In the cell fusion assays of the subject invention, a first cell that stably expresses an envelope protein of an enveloped virion on its surface, i.e., the “virion cell,” and a second cell that stably expresses a receptor for the envelope protein on its surface, i.e., the “target cell,” are employed. The virion and target cells each contain one component of a two component reporter system, e.g., a Tat based two component reporter system, that produces a detectable signal in the presence of cell fusion. In practicing the subject screening methods, the first and second cells are contacted with each other under cell fusion conditions in the presence of a candidate inhibitory agent. Next, the presence or absence of the detectable signal is detected. Finally, the inhibitory activity of the candidate agent is derived from the presence or absence of the detectable signal. Also provided are high throughput embodiments of the subject methods. In addition to the subject methods, systems and kits for performing the subject methods are provided.
REFERENCES:
Berger et al. (1995) “HIV envelope glycoprotein/CD4 interactions: studies using recombinant vaccinia virus vectors.” InHIV: A Practical Approach, J. Karn ed., IRL Press, vol. 2:123-145.
Blondell et al. (Feb. 25, 2000) Abstract from 3rdAnnual Conference on AIDS research in CA.
Broder et al. (1995) “Fusogenic selectivity of the envelope glycoprotein is a major determinant of human immunodeficiency virus type 1 tropism for CD4+ T-cell lines vs. primary macrophages.”Proc. Natl. Acad. Sci. USA, vol. 92:9004-9008.
Doranz et al. (1997) “A Small-molecule Inhibitor Directed against the Chemokine Receptor CSCR4 Prevents its Use as an HIV-1 Coreceptor.”J. Exp. Med., vol. 186(8):1395-1400.
Moir et al. (1996) “Expression of HIV env Gene in a Human T Cell Line for a Rapid and Quantifiable Cell Fusion Assay.”AIDS Research and Human Retroviruses, vol. 12:811-820.
Weiss et al. (1993) “Characterization of Stable Chinese Hamster Ovary Cells Expressing Wild-Type, Secreted, and Glycosylphosphatidylinositol-anchored Human Immunodeficiency Virus Type 1 Envelope Glycoprotein.”Journal of Virology, vol. 67(12):7060-7066.
Weiss et al. (1996) “Studies of HIV-1 envelope glycoprotein-mediated fusion using a simple fluorescence assay.”AIDS, vol. 10:241-246.
Yi et al. (1999) “Role of CXCR4 in Cell-Cell Fusion and Infection of Monocyte-Derived Macrophages by Primary Human Immunodeficiency Virus Type 1 (HIV-1) Strains: Two Distinct Mechanisms of HIV-1 Dual Tropism.”Journal of Virology, vol. 73(9):7117-7125.
Goldsmith Mark A.
You Yun
Borden Paula A.
Bozicevic Field & Francis LLP.
Field Bret E.
Park Hankyel T.
The Regents of the University of California
LandOfFree
Cell fusion assays for the identification of antiviral... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Cell fusion assays for the identification of antiviral..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Cell fusion assays for the identification of antiviral... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2850457