Methods for assaying inhibition of HIV-1 envelope glycoprotein-mediated membrane fusion

Abstract

This invention provides: agents determined to be capable of specifically inhibiting the fusion of a macrophage-tropic primary isolate of HIV-1 to a CD4+ cell, but not a T cell-tropic isolate of HIV-1 to a CD4+ cell; and agents determined to be capable of specifically inhibiting the fusion of a T cell-tropic isolate of HIV-1 to a CD4+ cell, but not a macrophage-tropic primary isolate of HIV-1 to a CD4+ cell. This invention also provides: agents capable of specifically inhibiting the fusion of a macrophage tropic primary isolate of HIV-1 with a CD+ cell susceptible to infection by a macrophage-tropic primary isolate of HIV-1; and agents capable of specifically inhibiting the fusion of a T cell-tropic isolate of HIV-1 with a CD4+ cell susceptible to infection by a T cell-tropic isolate of HIV-1. The agents include but are not limited to antibodies. This invention further provides: methods of inhibiting fusion of a macrophage-tropic primary isolate of HIV-1 with a CD+ cell susceptible to infection by a macrophage-tropic primary isolate of HIV-1 which comprises contacting the CD4+ cell with an amount of an agent capable of specifically inhibiting such fusion so as to thereby inhibit such fusion; and methods of inhibiting fusion of a T cell-tropic isolate of HIV-1 with a CD4+ cell susceptible to infection by a T cell-tropic isolate of HIV-1 which comprises contacting the CD4+ cell with an amount of an agent capable of specifically inhibiting such fusion so as to thereby inhibit such fusion.

Description

[0001] This application is a continuation-in-part application of U.S. Ser. No. 08 / 175,515, filed Jun. 7, 1995, the content of which is hereby incorporated into this application by reference.BACKGROUND OF THE INVENTION [0002] Throughout this application, various publications are referenced. The disclosure of these publications is hereby incorporated by reference into this application to describe more fully the art to which this invention pertains. [0003] HIV infects primarily helper T lymphocytes and monocytes / macrophages—cells that express surface CD4—leading to a gradual loss of immune function which results in the development of the human acquired immune deficiency syndrome (AIDS). The initial phase of the HIV replicative cycle involves the high affinity interaction between the HIV exterior envelope glycoprotein gp120 and the cellular receptor CD4 (Klatzmann, D. R., et al., Immunodef. Rev. 2, 43-66 (1990)). Following the attachment of HIV to the cell surface, viral and target cell...

AI Technical Summary

Problems solved by technology

This technique measures dye distribution rather than fluorescence intensity and as such cannot be performed using fluorometer.

The assay would not be easily automated and has not been performed with cells which stably express the HIV-1 envelope glycoprotein.

This assay takes at least 1 day to perform and cannot easily be adapted to new target cells such as primary macrophage cells.

This assay also does not measure cell fusion in real time and is thus not amenable to use in analyzing fusion kinetics.

It would be difficult to readily isolate sufficient quantities of clinical virus isolates to perform the assay.

Furthermore, this assay is more complicated and less reproducible than a RET assay using cells which stably express HIV-1 envelope glycoproteins and CD4.

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