Measurement of HIV virus load and genotypic resistance by gene amplification in asymptomatic subjects treated with combination therapy.
Quantification of viral load in HIV disease has become increasingly important as a marker of antiviral efficacy. We applied gene amplification techniques in vivo to asses antiretroviral activity of combination therapy. Five HIV-infected subjects, four of whom were drug naive, were administered combination therapy with zidovudine (ZDV) and didanosine (ddI). Plasma and peripheral blood mononuclear cells (PBMC) were obtained twice at baseline and then at 1, 3, 6, 9, and 12 months after the initiation of therapy. Results show that plasma HIV RNA copy number fell from 2,170 +/- 660/ml to undetectable at 1 month, with continued suppression at 12 months. HIV proviral DNA copy number decreased from 3.9 to 3.0 log10/10(6) CD4+ T cells at 12 months. Cell dilution cultures were positive in 4 of 5 subjects at baseline and in only 1 of 5 after 12 months. CD4+ T-cell count increased from 390 +/- 30/mm3 pretherapy, to 505 +/- 66/mm3 after 6 months of therapy, but returned to baseline levels after 12 months of therapy. No mutations were detected from PBMC DNA for codon 215 and 74 in the HIV pol gene from the drug-naive subjects. These findings suggest that gene amplification techniques can be used to study changes in viral load or genotype and can be applied in real time to samples from patients involved in clinical trials.
Holodniy
M
M
Center for AIDS Research, Stanford University, California.
Katzenstein
D
D
Winters
M
M
Montoya
J
J
Shafer
R
R
Kozal
M
M
Ragni
M
M
Merigan
T C
TC
D015496:CD4-Positive T-Lymphocytes; D003062:Codon; D016049:Didanosine / Q000008:administration & dosage*; D004359:Drug Therapy, Combination; D005784:Gene Amplification; D005814:Genes, Viral; D006678:HIV / Q000235:genetics*; D015658:HIV Infections / Q000188:drug therapy / Q000235:genetics* / Q000382:microbiology / Q000473:pathology; D006801:Humans; D007958:Leukocyte Count; D012367:RNA, Viral / Q000032:analysis*; D015215:Zidovudine / Q000008:administration & dosage*
D003062:Codon; D012367:RNA, Viral; D015215:Zidovudine; D016049:Didanosine