GB virus C (GBV-C) genomic RNA is present in specific lymphocytic cells in vivo

RUIZ V, GIORDANO M, MATHET V, RIVERO C, MINASSIAN M, CUESTAS M, FLEMING X, TRINKS J, OUBIÑA J

Pinamar, Argentina

Congreso; XLI Reunión Anual de SAIB; 2005

Sociedad Argentina de Investigación Bioquímica y Biología Molecular

INTRODUCTION: GBV-C -also known as Hepatitis G virus- is a human virus whose replication cell target has not been identified. OBJECTIVE: To investigate GBV-C cell tropism in peripheral blood mononuclear cells (PBMC) from a viremic blood donor. MATERIAL AND METHODS: Mononuclear and polymorphonuclear cells were obtained by standard procedures using Ficoll/Triyosom centrifugation and Dextran sedimentation. Monocytes were isolated by adherence to plastic and lymphoid subsets were purified using specific monoclonal antibodies and magnetic beads. RNA was extracted from cellular pellets and 1 mg was seeded for RT-Nested PCR for both genomic [+] and anti-genomic [-] strands by using primers specific for the 5 NCR of the virus. Amplicons were analyzed by agarosa gel electrophoresis and Southern hybridization with a 32P-labeled internal oligoprobe. RESULTS: GBV-C genomic RNA was consistently detected in B cells but not in monocytes, CD4 [+] T cells, polymorphonuclear cells, nor in the last wash from any cell population. CD8 [+] T cells proved positive in 2 of 4 assays. The anti-genomic GBV-C RNA (replicative form) was undetectable in all cell populations by using the highly strand-specific enzyme Tth DNA polymerase. CONCLUSIONS: GBV-C viral RNA is present in B and probably in CD8 [+] T cells in vivo, as shown for the first time in a natural infection.