Biological effects of bone marrow mesenchymal stem cells on hepatitis B virus in vitro

Abstract

The aim of the present study was to explore the effects of co-culturing bone marrow-derived mesenchymal stem cells (BM-MSCs) cultured with hepatitis B virus (HBV)-infected lymphocytes in vitro. BM-MSCs and lymphocytes from Brown Norway rats were obtained from the bone marrow and spleen, respectively. Rats were divided into the following five experimental groups: Group 1, splenic lymphocytes (SLCs); group 2, HepG2.2.15 cells; group 3, BM-MSCs + HepG2.2.15 cells; group 4, SLCs + HepG2.2.15 cells; and group 5, SLCs + BM-MSCs + HepG2.2.15 cells. The viability of lymphocytes and HepG2.2.15 cells was assessed using the MTT assay at 24, 48 and 72 h, respectively. Levels of supernatant HBV DNA and intracellular HBV covalently closed circular DNA (cccDNA) were measured using quantitative polymerase chain reaction. Supernatant cytokine levels were measured by enzyme-linked immunosorbent assay (ELISA). T cell subsets were quantified by flow cytometry using fluorescence-labeled antibodies. In addition, the HBV genome sequence was analyzed by direct gene sequencing. Levels of HBV DNA and cccDNA in group 5 were lower when compared with those in group 3 or group 4, with a significant difference observed at 48 h. The secretion of interferon-γ was negatively correlated with the level of HBV DNA, whereas secretion of interleukin (IL)-10 and IL-22 were positively correlated with the level of HBV DNA. Flow cytometry demonstrated that the percentage of CD3+CD8+ T cells was positively correlated with the levels of HBV DNA, and the CD3+CD4+/CD3+CD8+ ratio was negatively correlated with the level of HBV DNA. Almost no mutations in the HBV DNA sequence were detected in HepG2.2.15 cells co-cultured with BM-MSCs, SLCs, or in the two types of cells combined. BM-MSCs inhibited the expression of HBV DNA and enhanced the clearance of HBV, which may have been mediated by the regulation of the Tc1/Tc2 cell balance and the mode of cytokine secretion to modulate cytokine expression.

Current Knowledge of Th22 Cell and IL-22 Functions in Infectious Diseases

“…It has been confirmed that through the STAT3 pathway, IL-22 stimulates the proliferation of liver stem/progenitor cells (LPCs), cells participating in liver inflammatory responses [ 86 ]. Moreover, IL-22 production in hepatitis B patients is positively related to the HBV load [ 87 ]. By increasing CXCL9 and CXCL10 expression on hepatic stellate cells (HSCs), IL-22 also promotes inflammatory cell accumulation in the liver, leading to increased liver damage and hepatic fibrosis [ 88 ].…”

The Role of Interleukins in HBV Infection: A Narrative Review

“…Current evidence presents IL-22 as a double-edge sword in HBV infection. Studies support that serum concentration of IL-22 is elevated in CHB [57][58][59][60], and so is the Th17 cells' percentage in peripheral blood [58]. In mice, Feng et al showed that CD3+ Tcells induce liver progenitor cell proliferation via IL-22 expression [61], while Park et al demonstrated that IL-22 secretion may be a protective mechanism in preventing further liver injury, despite its positive association with serum ALT levels [62].…”

Mesenchymal Stromal Cells: an Antimicrobial and Host-Directed Therapy for Complex Infectious Diseases

“…However, MSCs can enhance host cellular response against both HBV and HCV in vitro. Human MSCs suppressed HBV replication in hepatoblastoma cells when cocultured with splenic lymphocytes and reduced HCV replication in epithelial cells (220,221).…”