Means and methods for monitoring protease inhibitor antiretroviral therapy and guiding therapeutic decisions in the treatment of HIV/AIDS
View Patent ↗This invention relates to antiviral drug susceptibility and resistance tests to be used in identifying effective drug regimens for the treatment of human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS), particularly treatment regimens including a protease inhibitor. The invention further relates to the means and methods of monitoring the clinical progression of HIV infection and its response to antiretroviral therapy using phenotypic or genotypic susceptibility assays.
1. A method for assessing the effectiveness of a candidate HIV antiretroviral compound, said method comprising:
a) culturing a host cell in the presence of said candidate HIV antiretroviral compound, wherein said host cell comprises a test vector comprising (i) a nucleic acid segment encoding HIV protease having a mutation at codon 88 and (ii) an indicator gene, wherein expression of said indicator gene depends upon the activity of said HIV protease;
b) measuring the amount of expression of said indicator gene in said host cell from step (a); and
c) comparing the amount of expression of said indicator gene as measured in step (b) with an amount of expression of said indicator gene measured in said host cell of step (a) cultured in the absence of said candidate HIV antiretroviral compound, wherein the difference between the amount of expression of said indicator gene measured in step (c) relative to that measured in step (b) correlates with the effectiveness of said candidate HIV antiretroviral compound, thereby assessing the effectiveness of said candidate HIV antiretroviral compound.
2. The method of claim 1 , wherein said a nucleic acid segment encoding HIV protease further comprises a mutation at codon(s) 63 and/or 77.
3. The method of claim 1 , wherein said a nucleic acid segment encoding HIV protease further comprises a mutation at codon(s) 63, 77 and/or 46, or a combination thereof.
4. The method of claim 1 , wherein said a nucleic acid segment encoding HIV protease further comprises a mutation at codon(s) 63, 77, 46, 10, 20 and/or 36, or a combination thereof.
5. A test vector comprising a nucleic acid segment encoding HIV protease having a mutation at codon 88, and an indicator gene, wherein expression of the indicator gene depends upon the activity of said HIV protease.
6. The test vector of claim 5 , wherein said nucleic acid segment encoding HIV protease further comprises a mutation at codon(s) 63 and/or 77.
7. The test vector of claim 5 , wherein said nucleic acid segment encoding HIV protease further comprises a mutation at codon(s) 63, 77 and/or 46, or a combination thereof.
8. The test vector of claim 5 , wherein said nucleic acid segment encoding HIV protease further comprises a mutation at codon(s) 63, 77, 46, 10, 20 and/or 36, or a combination thereof.