title: Phytohemagglutinin and concanavalin A activate hepatitis B virus in peripheral blood mononuclear cells of patients with chronic hepatitis B virus infection.
abstract: Peripheral blood mononuclear cells (PBMC) from 25 patients with chronic hepatitis B were tested for the presence of free monomeric hepatitis B virus (HBV) DNA migrating as a single 3.2 Kb band by Southern blot analysis. The PBMC were cultured for 7 days in the presence of phytohemagglutinin (PHA) or concanavalin A (ConA) both of which yielded a proliferative response. By contrast, both bacterial lipopolysaccharide (LPS) and interleukin 2 (IL2) failed to do so. Dot blot assays were used to monitor HBV DNA level increase within PBMC. Following mitogen exposure HBV DNA levels increased above pre-stimulation levels in 19/25 PHA cultures, 6/15 ConA cultures, 1/15 LPS cultures, and 1/15 IL2 cultures. In 15 patients, Southern blot analysis was carried out before and after PHA exposure. In 13/15 cases, a single 3.2 Kb band was observed in unstimulated cultures as well as in PHA cultures even though PHA induced a HBV DNA increase. One case exhibited bands migrating faster than the 3.2 Kb signal, compatible with replicating intermediates and one case provided evidence of viral concatemers within PBMC after PHA stimulation. No HBV DNA was detected in the culture supernatants. The increase of HBV DNA level in PBMC induced by mitogen was strongly associated with an increase in HBV DNA expression (HBV RNA and HBs antigen). These studies indicate that HBV DNA present in human PBMC does represent a potential reservoir for infection with endogenous reactivation following PBMC activation.
authors.affiliation: INSERM U271, Lyon, France.
authors.forename: P
authors.initials: P
authors.lastname: Bouffard
authors.forename: J P
authors.initials: JP
authors.lastname: Lamelin
authors.forename: F
authors.initials: F
authors.lastname: Zoulim
authors.forename: D
authors.initials: D
authors.lastname: Lepot
authors.forename: C
authors.initials: C
authors.lastname: Trepo
mesh_terms: D015139:Blotting, Southern; D002908:Chronic Disease; D003208:Concanavalin A / Q000494:pharmacology; D003470:Culture Media; D004279:DNA, Viral / Q000097:blood*; D005260:Female; D006509:Hepatitis B / Q000382:microbiology; D006514:Hepatitis B Surface Antigens / Q000097:blood; D006515:Hepatitis B virus / Q000235:genetics* / Q000254:growth & development; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000382:microbiology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008934:Mitogens / Q000494:pharmacology*; D010835:Phytohemagglutinins / Q000494:pharmacology; D012367:RNA, Viral / Q000097:blood
chemical_list: D003470:Culture Media; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D007376:Interleukin-2; D008070:Lipopolysaccharides; D008934:Mitogens; D010835:Phytohemagglutinins; D012367:RNA, Viral; D003208:Concanavalin A