Detection of HHV-8 (human herpesvirus-8) genomes in induced peripheral blood mononuclear cells (PBMCs) from US blood donors

• Published in: Vox sanguinis 2011-04, Vol.100 (3), p.267-271
• Main Authors: Qu, L., Triulzi, D. J., Rowe, D. T., Jenkins, F. J.
• Format: Article
• Language: English
• Subjects: Antibodies; B-Lymphocytes - virology; Blood & organ donations; Blood Cells; Blood Donors; Blood products; Cell culture; Cell Culture Techniques; Cellular biology; DNA, Viral - blood; Genome, Viral; Genomes; Herpes viruses; Herpesvirus 8, Human - isolation & purification; Human herpesvirus 8; Humans; induction of PBMC; Kaposi's sarcoma; Leukocytes, Mononuclear - virology; Lymphocytes B; Media (culture); Nitrogen; Peripheral blood mononuclear cells; phorbol acetate; Polymerase chain reaction; Purification; United States
• ISSN: 0042-9007; 1423-0410
• DOI: 10.1111/j.1423-0410.2010.01404.x

Background  Human herpesvirus‐8 (HHV‐8) causes Kaposi’s sarcoma and can be detected and induced in peripheral blood mononuclear cells (PBMCs) from infected individuals. The prevalence of viral genomes in induced/cultured PBMCs from healthy blood donors has not been systematically studied. Materials and Methods  PBMCs from 164 donors were purified and stored as two equal aliquots in liquid nitrogen. One aliquot was used for CD19+ B‐cell purification with a fraction reserved for DNA extraction. The second aliquot was cultured for 2 or 4 days in culture media containing n‐butyrate and tetradecanoyl phorbol acetate. DNA was extracted from all four cell sources: PBMCs, purified B cells, induced PBMCs harvested at days 2 and 4 of culture. A sensitive real‐time PCR with a DNA equivalent of 3 × 105 cells per reaction was run in duplicate for all samples along with a quantitative HHV‐8 DNA standard ranging from 1·6 to 200 copies. Results  For all 164 donors, HHV‐8 genomes were not detected in the DNA equivalent of 3–6 × 105 of PBMCs and induced/cultured PBMCs with a real‐time PCR assay (95% CI: 0–3·5/164). HHV‐8 DNA was not detected from DNA equivalent of 1·5 (0·5–5·6) × 105 CD19+ B cells from 139/164 donors. HHV‐8 antibodies were detected in 7 of the 164 donors (4·3%). Conclusions  HHV‐8 genomes were not detected from PBMCs, induced/cultured PBMCs and CD19+ B cells from 164 blood donors. The level of detectable HHV‐8 genomes in blood donors seems to be extremely low, if they exist.