Selective detection of human hepatitis B virus surface and core antigens in some peripheral blood mononuclear cell subsets by flow cytometry.
The presence of HBs and HBc antigens was investigated, by flow cytometry, on the surface of peripheral mononuclear cells (PBMC) from the following phenotype: CD3 (T lymphocytes), CD4 (T helper/inducer), CD8 (T cytotoxic/suppressor), CD19 (B lymphocytes) and CD56 (NK cells) among 8 patients suffering from chronic hepatitis B and 5 healthy HBV-negative subjects. This study demonstrated the presence of HBsAg and HBcAg on the lymphocyte surface for most of the patients. The mean percentage of labelled cells was 17% for HBsAg and 15% for HBcAg. Among the different lymphocyte subsets only B lymphocytes and the NK cells expressed HBsAg for 57% and 26% of cells, respectively. Similarly HBcAg was also detected among CD19 and CD56 cells only. PCR was used to search for the presence of HBV DNA and RNA in PBMC, using primers located in the S gene. HBV DNA was detected with variable intensity in the CD3, CD4, CD19 and CD56 subsets following their separation with a cell sorter. For HBV RNA the signal obtained after PCR and Southern blotting was higher for CD56 and CD19 cells than for CD3 cells and undetectable for CD4 cells. This study demonstrates that replication and transcription can occur in CD19 and CD56 cells. Positive signals in CD3 cells may possibly be due to contamination of this subpopulation by NK cells.
Chemin
I
I
INSERM U271, Lyon, France.
Vermot-Desroches
C
C
Baginski
I
I
Saurin
J C
JC
Laurent
F
F
Zoulim
F
F
Bernaud
J
J
Lamelin
J P
JP
Hantz
O
O
Rigal
D
D
D015703:Antigens, CD / Q000097:blood; D001402:B-Lymphocytes / Q000276:immunology; D015139:Blotting, Southern; D002908:Chronic Disease; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006509:Hepatitis B / Q000097:blood / Q000276:immunology*; D006512:Hepatitis B Core Antigens / Q000097:blood*; D006514:Hepatitis B Surface Antigens / Q000097:blood*; D006801:Humans; D016130:Immunophenotyping; D007694:Killer Cells, Natural / Q000276:immunology; D016131:Lymphocyte Subsets / Q000276:immunology*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis
D015703:Antigens, CD; D006512:Hepatitis B Core Antigens; D006514:Hepatitis B Surface Antigens; D012367:RNA, Viral