Expression of CD3+ CD8+ human leukocyte antigen-A2+ T lymphocytes with specificity to the different hepatitis B virus peptides in patients with hepatitis B associated hepatocellular carcinoma
10.3760/cma.j.issn.1000-6680.2012.05.004
- VernacularTitle:乙型肝炎相关肝癌患者乙型肝炎病毒不同抗原肽特异性CD3+CD8+人白细胞抗原-A2+细胞表达
- Author:
Jilin CHENG
;
Liwei WANG
;
Chenli QIU
;
Yingchun AI
;
Jihua LU
;
Keshan YIN
;
Shaoping HUANG
;
Rong TANG
;
Lie XU
;
Yi ZHANG
- Publication Type:Journal Article
- Keywords:
Carcinoma,hepatocellular;
Monocytes;
Hepatitis B virus;
HLA-A2 antigen;
Antigens,CD3;
Antigens,CD8;
Peptides
- From:
Chinese Journal of Infectious Diseases
2012;30(5):264-267
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the expression of CD3+ CD8+ human leukocyte antigen (HLA)-A2+T lymphocytes with specificity to the different hepatitis B virus (HBV) peptides in the peripheral blood mononuclear cells (PBMC)from the patients with hepatitis B associated hepatocellular carcinoma (HCC).MethodsThe HLA-A2+ PBMC from four patients with hepatitis B associated HCC were incubated with five HBV/HLA-A2 pentamers respectively,which were HBV sAg (FLLTRILTI),HBV sAg (GLSPTVWLSV),HBV sAg (WLSLLVPFV),HBV core (FLPSDFFPSV),and HBV pol (FLLSLGIHL),as well as anti-CD3-pacific blue and anti-CD8-fluorescein isothiocyanate (FITC).Then,HBV/HLA-A2-CD3-CD8 positive cells were detected by flow cytometry. The monoclonal HBV/HLA-A2-CD3-CD8+ cells were acquired by fluorescenceactivated cell sorter,and cultured and identified by flow cytometry.The anti-HBV specific T lymphocytes were then cultured with HepG2 (HLA-A2+ ) cells and the release of interferon γ (IFN-γ)were determined by enzyme-linked immunosorbent assay (ELISA),Res(a)ltsThe percentage of antiHBV T lymphoeytes with specificity to GLSPTVWLSV in total CD8+ T lymphoeytes from four patients with hepatitis B associated HCC was 1.44％±0.04％,which was higher than those to other four HBV antigen peptides (0.68％±0.08％ of FLLTRILTI,1.06％±0.09％ of FLPSDFFPSV,0.56％ ±0.04％ of FLLSLGIHL,and 0.46％ ±0.08％ of WLSLLVPFV) (t=0.001,P＜0.05).The two lines of monoclonal cell with specificity to GLSPTVWLSV both exhibited high level of IFN-γ expression after incubated with hepatic carcinoma cell line HepG2 (HLA-A2+)with HBV GLSPTVWLSV peptide.ConclusionsCD3+ CD8+ HLA-A2+ cells with specificity to the different HBV peptides exist in PBMC of patients with hepatitis B associated HCC.The expression level depends on HBV antigen peptide sequences and genomic sites.
