Human Herpesvirus 8 DNA Load in Leukocytes of Human Immunodeficiency Virus-Infected Subjects: Correlation with the Presence of Kaposi’s Sarcoma and Response to Anticytomegalovirus Therapy

Abstract

Specific human herpesvirus 8 (HHV-8) DNA sequences were found in leukocytes of 12 of 29 (41.4%) AIDS subjects with Kaposi's sarcoma (KS), whereas they were found in 4 of 43 (9.3%) AIDS subjects without KS (P ‫؍‬ 0.003), although the peak HHV-8 DNA load in PCR-positive subjects with KS (mean, 425 copies per 0.2 g of DNA) did not significantly differ from the one found in PCR-positive patients without KS (mean, 218 copies). The use of intravenous ganciclovir or foscarnet therapy to treat cytomegalovirus disease did not affect the HHV-8 DNA load in seven patients for whom serial samples were analyzed.

Evaluation of the human herpesvirus 8 DNA load in blood and Kaposi's sarcoma skin lesions from AIDS Patients on highly active antiretroviral therapy

“…In contrast, others have detected expression of two replicative genes using reverse transcription±PCR in KS lesions but not in PBMC of patients with KS [3]. In agreement with the last study, we reported previously that ganciclovir and foscarnet (two viral DNA polymerase inhibitors shown to inhibit HHV-8 in vitro [12]) had no immediate effects in reducing the HHV-8 DNA burden in leukocytes suggesting that most of the viral genome detected in the blood of patients with KS was from latent, non-replicative viruses [9].…”

“…Samples that contained high HHV-8 copy numbers were diluted and retested to ®t into the standard curve. The presence of the cellular B-actin gene was also con®rmed by PCR in all HHV-8 PCR-negative samples [9].…”

“…Two speci®c 59-biotinylated probes were used to differentiate between HHV-8 and the competitor DNA target [9]. The probes (1.5 pmol) were hybridized for 3 h at 558C with the denatured digoxigenin-labelled PCR products, then hybrids were captured on a streptavidin-coated microtiter plate using different wells for the two probes.…”

“…Skin biopsies (about 25 mg) were processed using the QIAmp tissue kit (QIAGEN, Mississauga, ON, Canada) and the amount of cellular DNA was evaluated by spectrophotometry. The equivalent of 0.2 ìg DNA from PBMC and skin samples was used in a quantitative-competitive PCR assay as reported previously [9] except that 200 ìM of a PCR labelling mix including digoxigenin-11-dUTP (Boehringer Mannheim, Laval, Canada) was incorporated into the PCR mixture. Serial dilutions of an external linearized DNA standard were also ampli®ed in the presence of 100 copies of an internal DNA competitor [9] and 0.2 ìg of HHV-8-negative blood or skin DNA background in order to construct a standard curve for each PCR assay.…”

A Mathematical Model for the Treatment of Aids-Related Kaposi's Sarcoma

“…For example, HHV-8 DNA plasma load in longitudinal samples from HIV-KS patients treated with HAART showed both increasing and decreasing trends 20 while treatment with antiherpetic drugs did not affect HHV-8 load in PBMCs. 21 Interestingly, no correlation was found between HHV-8 load and progression to KS, or disease stage in a longitudinal analysis of serum HHV-8 DNA load in AIDS-KS patients. 22 To our knowledge, the work of Nani and Jin 23 is one of the few published mathematical studies modeling HAART for HIV-KS dynamics.…”