title: Infected hematopoietic stem cells and with integrated HBV DNA generate defective T cells in chronic HBV infection patients.
abstract: A weak T-cell response plays a key role in the persistence of hepatitis B virus (HBV) infection. We aimed to confirm that T-cell defects in patients with chronic HBV infection are associated with HBV DNA infection of bone marrow (BM) hematopoietic stem cells (HSCs). Using reverse transcription polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), we observed the transcription of HBsAg coding genes and confirmed the integration of HBV DNA in CD34(+) BM HSCs from chronic HBV infection patients. T cells were generated by coculturing the HSCs with delta-like ligand 1-expressing OP9 (OP9-DL1) cells. The phenotypes of the T cells were then evaluated by flow cytometric (FACS) analysis on days 14 and 25. The results demonstrated that fewer CD3(+) TCRaβ(+) CD3(+) CD4(+) and CD4(+) CD8(+) T cells were generated from the HSCs of the patients than from the healthy controls, (P < 0.01) but the frequency of CD3(+) D8(+) T cells was not significantly different between the two group (P > 0.05). In contrast, CD4(+) CD25(+) T cells were more in the patient group than in healthy controls (P < 0.01) on both days 14 and 25. There were fewer CD3(+) CD4(+) /CD3(+) CD8(+) cells in the patient group than in the healthy control group on day 25 (P < 0.05). Less proliferation and lower levels of IL-2 and IFN- γ were also observed in the patient group compared with the control group (P < 0.05).These data suggest that HBV DNA infected and integrated into the BM HSCs from patients with chronic HBV infection and that these BM HSCs generated defective T cells.
authors.affiliation: Department of Infectious Disease, the First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Shi
authors.forename: Y
authors.initials: Y
authors.lastname: Lan
authors.forename: F
authors.initials: F
authors.lastname: Cao
authors.forename: Y
authors.initials: Y
authors.lastname: Teng
authors.forename: L
authors.initials: L
authors.lastname: Li
authors.forename: F
authors.initials: F
authors.lastname: Wang
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: J
authors.initials: J
authors.lastname: Zhou
authors.forename: Y
authors.initials: Y
authors.lastname: Li
mesh_terms: D015703:Antigens, CD / Q000032:analysis; D002478:Cells, Cultured; D018920:Coculture Techniques; D004279:DNA, Viral / Q000302:isolation & purification*; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000821:virology*; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000235:genetics; D019698:Hepatitis C, Chronic / Q000473:pathology / Q000821:virology*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016131:Lymphocyte Subsets / Q000737:chemistry / Q000276:immunology* / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000737:chemistry / Q000276:immunology* / Q000821:virology*
chemical_list: D015703:Antigens, CD; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens
keywords: HBV DNA; HSCs; T-cell defects; integration; replication