title: Kinetics of phytohemaglutinin-induced IFN-gamma and TNF-alpha expression in peripheral blood mononuclear cells from patients with chronic hepatitis B after liver transplantation.
abstract: OBJECTIVE
  To study the association between host immunity and hepatitis B virus (HBV) recurrence after liver transplantation.
  
  
  METHODS
  Peripheral blood mononuclear cells (PBMC) were isolated from 40 patients with hepatitis B and underwent orthotopic liver transplantation (OLT) before and 2, 4, 8 wk after surgery. After being cultured in vitro for 72 h, the levels of INF-gamma and TNF-alpha in culture supernatants were detected with ELISA. At the same time, the quantities of HBV DNA in serum and PBMCs were measured by real time PCR.
  
  
  RESULTS
  The levels of INF-gamma and TNF-alpha in PBMC culture supernatants decreased before and 2, 4 wk after surgery in turns (INF-gamma 155.52+/-72.32 ng/L vs 14.76+/-9.88 ng/L vs 13.22+/-10.35 ng/L, F = 6.946, P = 0.027 < 0.05; TNF-alpha 80.839+/-46.75 ng/L vs 18.59+/-17.29 ng/L vs 9.758+/-7.96 ng/L, F = 22.61, P = 0.0001 < 0.05). The levels of INF-gamma and TNF-alpha were higher in groups with phytohemagglutinin (PHA) than in those without PHA before surgery. However, the difference disappeared following OLT. Furthermore, INF-gamma and TNF-alpha could not be detected in most patients at wk 4 and none at wk 8 after OLT. The HBV detection rate and virus load in PBMC before and 2, 4 wk after surgery were fluctuated (HBV detected rate: 51.4%, 13.3%, 50% respectively; HBV DNA: 3.55+/-0.674 log10 copies/mL vs 3.00+/-0.329 log10 copies/mL vs 4.608+/-1.344 log10 copies/mL, F = 7.582, P = 0.002 < 0.05). HBV DNA in serum was 4.48+/-1.463 log10 copies/mL before surgery and <10(3) copies/mL after OLT except for one with 5.72 x 10(6) copies/mL 4 wk after OLT who was diagnosed as HBV recurrence. The levels of INF-gamma and TNF-alpha were lower in patients with a high HBV load than in those with a low HBV load (HBV DNA detected/undetected in PBMCs: IFN-gamma 138.08+/-72.44 ng/L vs 164.24+/-72.07 ng/L, t = 1.065, P = 0.297 > 0.05, TNF-alpha 80.75+/-47.30 ng/L vs 74.10+/-49.70 ng/L, t = 0.407, P = 0.686 > 0.05; HBV DNA positive/negative: IFN-gamma 136.77+/-70.04 ng/L vs 175.27+/-71.50 ng/L, t = 1.702, P = 0.097 > 0.05; TNF-alpha 75.37+/-43.02 ng/L vs 81.53+/-52.46 ng/L, t = 0.402, P = 0.690 > 0.05).
  
  
  CONCLUSIONS
  The yielding of INF-gamma and TNF-alpha from PBMCs is inhibited significantly by immunosuppressive agents following OLT with HBV load increased, indicating that the impaired immunity of host is associated with HBV recurrence after OLT.
authors.affiliation: Department of Gastroenterology, the First Affiliated Hospital of Sun-Yat Sen University, Guangzhou 510080, Guangdong Province, China.
authors.forename: Ying-Mei
authors.initials: YM
authors.lastname: Tang
authors.forename: Min-Hu
authors.initials: MH
authors.lastname: Chen
authors.forename: Gui-Hua
authors.initials: GH
authors.lastname: Chen
authors.forename: Chang-Jie
authors.initials: CJ
authors.lastname: Cai
authors.forename: Xiao-Shun
authors.initials: XS
authors.lastname: He
authors.forename: Min-Giang
authors.initials: MG
authors.lastname: Lu
authors.forename: Wei-Min
authors.initials: WM
authors.lastname: Bao
mesh_terms: D002478:Cells, Cultured; D019694:Hepatitis B, Chronic / Q000276:immunology* / Q000601:surgery*; D006801:Humans; D007165:Immunosuppression Therapy / Q000009:adverse effects; D007371:Interferon-gamma / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism*; D016031:Liver Transplantation; D010835:Phytohemagglutinins / Q000494:pharmacology; D012008:Recurrence; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D010835:Phytohemagglutinins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma