[Detection of HBV specific cytotoxic lymphocytes responses in severe hepatitis B patients using MHC-peptide tetramer flow cytometry].
OBJECTIVE
To explore the role played by HBV antigen specific cytotoxic lymphocytes (CTLs) in severe hepatitis B patients.


METHODS
Three kinds of HBV specific CD8+ T cells responding to HBV core 18-27, polymerase 575-583 and envelope 335-343 specific CD8+ T cells were searched with major histocompatibility complex (MHC)-peptide tetramer flow cytometry in the patients we studied. The cytokines produced by these specific CTLs such as IFNgamma, TNFalpha, IL-4, IL-10 were detected using ELISPOT assays. Their ability to lyse target cells was analyzed using a CytoTox96 Non-Radioactive Cytotoxicity Assay kit (Promega).


RESULTS
The number of HBV core 18-27 specific CD8+ T cells was significantly higher in the peripheral blood of acute severe hepatitis B group patients than that in chronic severe hepatitis group (P less than 0.05), but lower than that in the acute hepatitis B patients. The IFNgamma and TNFalpha levels of acute severe hepatitis B patients were both significantly higher than those in the chronic severe hepatitis group (P less than 0.05). The ability to lyse target cells of HBV core 18-27 CTLs was also significantly higher in the acute severe hepatitis B group than that in the chronic severe hepatitis B group (P less than 0.05).


CONCLUSIONS
The response to antigen stimulation was much higher in acute severe hepatitis B than in chronic severe hepatitis B patients. The specific CTLs persisted among the peripheral blood mononuclear cells in acute severe hepatitis B, which may be related to the viral clearance.
Chen
Yu
Y
Department of Infectious Diseases, First Hospital of Zhejiang University, Hangzhou 310003, China.
Wu
Wei
W
Li
Lan-juan
LJ
D000293:Adolescent; D000328:Adult; D000368:Aged; D005260:Female; D005434:Flow Cytometry; D006509:Hepatitis B / Q000276:immunology* / Q000821:virology; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D055815:Young Adult