Method for testing drug susceptibility of HIV
AssigneeMUSC Foundation for Research Development
InventorJian-yun Dong
Life of this patent
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Abstract
Methods, compositions and kits are provided for testing susceptibility of HIV to drug treatment, such as drug resistance of HIV and inhibition of HIV replication by a drug candidate. In one aspect of the invention, a method is provided for detecting drug resistance of HIV contained in a sample from an individual infected with HIV. In one embodiment, the method employs an indicator cell line which over-expresses CD4 and one or more co-receptors for HIV such as CXCR4 and CCR5 at high levels to render the cells susceptible to productive infection of various strains, subtypes or clades of HIV from both laboratory and clinical isolates. The methods, compositions and kits can be used for high throughput screening of HIV patient samples, anti-HIV agents, and for designing customized HIV therapy.
Claims61
A method for detecting drug resistance of HIV in a sample, comprising:
- taking a culture of recombinant cells in which at least one of the recombinant cells comprises
- a reporter sequence comprising a reporter gene whose expression is regulated by a protein specific to HIV, CD4, and
- one or more cell surface co-receptors for HIV, wherein the one or more cell surface co-receptors are each encoded by a heterologous sequence and expressed at an elevated level relative to the level of the corresponding cell surface co-receptor naturally expressed in a human cell such that productive infection of the recombinant cell by HIV is achieved, which is defined by HIV viral replication and the infection of non-infected cells in the culture of the recombinant cells;
- contacting the cell culture with a first sample containing HIV;
- adding an anti-HIV agent to the cell culture;
- and
- detecting a change in a level of expression of the reporter gene in the cells in the culture.
A method for detecting drug resistance of HIV in a sample, comprising:
- taking a first cell culture containing CD4 and one or more cell surface co-receptors for HIV at sufficient levels such that productive infection by HIV is achieved;
- contacting the first cell culture with a first sample containing HIV;
- adding an anti-HIV agent to the first cell culture;
- incubating the first culture in the presence of the first sample and the anti-HIV agent for a suitable period time;
- taking a second cell culture containing a reporter gene whose expression is regulated by a protein specific to HIV, CD4 and one or more cell surface co-receptors for HIV at sufficient levels such that productive infection by HIV is achieved;
- transferring the supernatant of the first cell culture after the incubation to the second cell culture;
- and
- detecting a change in a level of expression of the reporter gene in the cells in the second cell culture.
Dependencies are inferred from the claim text.