title: Quantitation of human immunodeficiency virus type 1 DNA forms with the second template switch in peripheral blood cells predicts disease progression independently of plasma RNA load.
abstract: There are several forms of human immunodeficiency virus type 1 (HIV-1) DNA in peripheral blood T cells and lymph nodes in untreated HIV-1-infected individuals and in patients whose plasma HIV-1 RNA levels are suppressed by long-term combination antiretroviral therapy. However, it remains to be established whether the concentration of HIV-1 DNA in cells predicts the clinical outcome of HIV-1 infection. In this report, we measured the concentration of HIV-1 DNA forms which has undergone the second template switch (STS DNA) and 2-long-terminal-repeat DNA circles in peripheral blood mononuclear cell (PBMC) samples. To do this, we used molecular-beacon-based real-time PCR assays and studied 130 patients with hemophilia in the Multicenter Hemophilia Cohort Study. We assessed the influence of baseline HIV-1 STS DNA levels on the progression of HIV-1 disease in the absence of combination antiretroviral therapy by Kaplan-Meier and Cox regression analysis. Among the patients who progressed to AIDS, the median levels (interquartile ranges) of STS HIV-1 DNA in PBMC were significantly higher than those of patients who remained AIDS free during the 16 years of follow-up (1,017 [235 to 6,059] and 286 [31 to 732] copies per 10(6) PBMC, respectively; P < 0.0001). Rates of progression to death and development of AIDS varied significantly (log rank P < 0.001) by quartile distribution of HIV-1 STS DNA levels. After adjustment for age at seroconversion, baseline CD4(+) T-cell counts, plasma viral load, and T-cell-receptor excision circles, the relative hazards (RH) of death and AIDS were significantly increased with higher HIV-1 STS DNA levels (adjusted RH, 1.84 [95% confidence interval (CI), 1.30 to 2.59] and 2.62 [95% CI, 1.75 to 3.93] per 10-fold increase per 10(6) PBMC, respectively). HIV-1 STS DNA levels in each individual remained steady in longitudinal PBMC samples during 16 years of follow-up. Our findings show that the concentration of HIV-1 STS DNA in PBMC complements the HIV-1 RNA load in plasma in predicting the clinical outcome of HIV-1 disease. This parameter may have important implications for understanding the virological response to combination antiretroviral therapy.
authors.affiliation: Department of Hygiene and Epidemiology, Athens University Medical School, 75 Mikras Asias, 11527 Athens, Greece. lkostrik@med.uoa.gr
authors.forename: Leondios G
authors.initials: LG
authors.lastname: Kostrikis
authors.forename: Giota
authors.initials: G
authors.lastname: Touloumi
authors.forename: Rose
authors.initials: R
authors.lastname: Karanicolas
authors.forename: Nikos
authors.initials: N
authors.lastname: Pantazis
authors.forename: Cleo
authors.initials: C
authors.lastname: Anastassopoulou
authors.forename: Anastasia
authors.initials: A
authors.lastname: Karafoulidou
authors.forename: James J
authors.initials: JJ
authors.lastname: Goedert
authors.forename: Angelos
authors.initials: A
authors.lastname: Hatzakis
mesh_terms: D004279:DNA, Viral / Q000097:blood*; D018450:Disease Progression; D015658:HIV Infections / Q000097:blood / Q000821:virology*; D015497:HIV-1 / Q000235:genetics*; D006467:Hemophilia A / Q000097:blood / Q000150:complications / Q000821:virology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods; D011237:Predictive Value of Tests; D012367:RNA, Viral / Q000097:blood*; D013698:Templates, Genetic; D019562:Viral Load
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral