HBV viral load within subpopulations of peripheral blood mononuclear cells in HBV infection using limiting dilution PCR.
Extrahepatic viral load in peripheral blood mononuclear cells (PBMCs) of patients with hepatitis B virus (HBV) is still under debate. In this study, HBV infection rates and viral titers were examined within all PBMC subpopulations using limiting dilution-PCR (LD-PCR). PBMCs of patients with acute or chronic hepatitis B were separated by magnetic beads in monocytes, B-cells, CD4+ T-cells, CD8+ T-cells, and NK cells. Using two-round nested PCR, HBV-DNA sequences were detected in all patients examined within each PBMC subpopulation. The frequencies of HBV-positive cells and viral loads were calculated by Poisson analysis of HBV PCR results from serial dilutions of cells and cell lysates. Highest infection rates were found in monocytes and B-cells followed by CD8 + T-cells, NK cells, and CD4+ T-cells. Concerning all subsets, frequencies of HBV-positive cells were 50- to 500-fold higher in chronic than in acute hepatitis B. Viral loads were mostly estimated at about one HBV genome per HBV-positive cell. Moreover, slightly elevated HBV titers were seen in B-cells, CD4+ T-cells, and NK cells in both acute and chronic hepatitis B. It was demonstrated that beside a generally more latent HBV infection in PBMCs, elevated HBV titers point to replication or selective viral uptake within particular PBMC subsets. Therefore, the data suggest that HBV-infected PBMCs may participate in persistence of HBV.
Trippler
M
M
Uniklinik Essen, Abteilung für Gastroenterologie und Hepatologie, Essen, Germany.
Meyer zum Büschenfelde
K H
KH
Gerken
G
G
D004279:DNA, Viral / Q000032:analysis; D005260:Female; D005914:Globins / Q000235:genetics; D006509:Hepatitis B / Q000276:immunology / Q000821:virology*; D006515:Hepatitis B virus / Q000302:isolation & purification* / Q000502:physiology*; D019694:Hepatitis B, Chronic / Q000276:immunology / Q000821:virology; D006801:Humans; D018189:Immunomagnetic Separation; D007963:Leukocytes, Mononuclear / Q000145:classification / Q000821:virology*; D008297:Male; D016012:Poisson Distribution; D016133:Polymerase Chain Reaction / Q000379:methods*; D019562:Viral Load
D004279:DNA, Viral; D005914:Globins