AIM: To find new serum biomarkers for liver cirrhosis (LC) in chronic carriers of hepatitis B virus (HBV).METHODS: Surface enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectrometry was used to ...AIM: To find new serum biomarkers for liver cirrhosis (LC) in chronic carriers of hepatitis B virus (HBV).METHODS: Surface enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectrometry was used to discover biomarkers for differentiating HBV induced LC from non-cirrhotic cohorts. A training population of 25 patients with HBV-induced LC, 20 patients with HCC, and 25 closely age-matched healthy men, was studied.RESULTS: Two biomarkers with Mr 7 772 and 3 933 were detected in sera of non-cirrhotic cohorts, but not in patients with HBV-induced LC. A sensitivity of 80% for all LC patients,a specificity of 81.8% for all non-cirrhotic cohorts and a positive predictive value of 75% for the study population were obtained.CONCLUSION: These two serum biomarkers for HBV-induced LC might be used for diagnosis and assessment of disease progression.展开更多
AIM:Gp96,also known as Grp94,is a member of heat shock protein (HSP) family and binds repertoires of peptides thereof eliciting peptide-specific T cell immune responses. It predominantly locates inside the endoplasmic...AIM:Gp96,also known as Grp94,is a member of heat shock protein (HSP) family and binds repertoires of peptides thereof eliciting peptide-specific T cell immune responses. It predominantly locates inside the endoplasmic reticulum (ER) with some cell surface expression in certain cancerous cells.Previous studies have shown that gp96 expression level was up-regulated in tumor cells,including hepatocellular carcinoma (HCC).However,relationship between the extent of gp96 expression and disease progression especially HBV-induced chronic infection,cirrhosis and hepatocellular carcinoma,has not been addressed before.As primary HCC can be induced and progressed from chronic hepatitis B virus (HBV) infection and HBV-induced cirrhosis,we designed an immunohistochemical experiment to test the correlation between gp96 expression level and HBV-induced disease progression,from chronic HBV infection,cirrhosis to HCC. METHODS:We chose liver samples from different patients of hepatitis B virus induced diseases,including chronic hepatitis B (77 patients),cirrhosis (27 patients) and primary HCC (30 patients),to test the expression level of gp96 in different affected groups.Formalin-fixed,and paraffin- embedded liver tissues taken from these patients were immuno-stained by using an anti-gp96 monoclonal antibody for the expression level of gp96 protein in the sections.In addition,Western blotting of whole cell lysates derived from established human embryonic liver cell lines and several human HCC cell lines (Huh7,HepG2,SSMC-7721) was compared with the expression of gp96. RESULTS:We found that the extent of elevated gp96 expression was significantly correlated with the disease progression,and was the highest in HCC patients,lowest in chronic HBV infection and was that of the cirrhosis in the middle. CONCLUSION:Increased expression of gp96 might be used as a diagnostic or prognostic bio-marker for the HBV infection and HBV-induced diseases.展开更多
AIM: Most studies on the immune effect of gp96 were focused on its enhancement of CTLs. It is interesting to know whether gp96 could influence the humoral immune response, and whether the recombinant N-terminal fragme...AIM: Most studies on the immune effect of gp96 were focused on its enhancement of CTLs. It is interesting to know whether gp96 could influence the humoral immune response, and whether the recombinant N-terminal fragment of gp96 could substitute native gp96 to stimulate the immune system.METHODS: gp96 isolated from livers of normal mice and its N-terminal fragment (amino acid 22-355) expressed in E coli were used for immunization of BALb/c mice. Eight groups of mice received one of the following regiments subcutaneously in 100 μL phosphate buffered saline (PBS)at an interval of 3 wk. Group 1: PBS only; group 2:gp96 only; group 3: N-terminal fragment only; group 4: HBsAg only; group 5: HBsAg+gp96; group 6: HBsAg+N-terminalfragment; group 7: HBsAg+incomplete Freud's adjuvant; group 8: HBsAg+N-terminal fragment (95 ℃ heated for 30 min). Serum anti-HBsAg antibody levels were assayed by ELISA. CTL responses in splenocytes were analyzed by ELISPOT after the last vaccination.RESULTS: The average titer of serum anti-HBsAg antibodyin the mice immunized with HBsAg together with gp96 or its N-terminal fragment were much higher than those immunized with HBsAg alone detected by ELISA. The cellular immune response of the mice immunized with HBsAg together with gp96 or its N-terminal fragment was not different with those immunized with HBsAg alone measured by ELISPOT assay.CONCLUSION: gp96 or its N-terminal fragment greatly improved humoral immune response induced by HBsAg, but failed to enhance the CTL response, which demonstrated the potential of using gp96 or its N-terminal fragment as a possible adjuvant to augment humoral immune response against HBV infection.展开更多
基金Supported by the Major State Basic Research Development Program of China(973 Program),No.2001CB510001
文摘AIM: To find new serum biomarkers for liver cirrhosis (LC) in chronic carriers of hepatitis B virus (HBV).METHODS: Surface enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectrometry was used to discover biomarkers for differentiating HBV induced LC from non-cirrhotic cohorts. A training population of 25 patients with HBV-induced LC, 20 patients with HCC, and 25 closely age-matched healthy men, was studied.RESULTS: Two biomarkers with Mr 7 772 and 3 933 were detected in sera of non-cirrhotic cohorts, but not in patients with HBV-induced LC. A sensitivity of 80% for all LC patients,a specificity of 81.8% for all non-cirrhotic cohorts and a positive predictive value of 75% for the study population were obtained.CONCLUSION: These two serum biomarkers for HBV-induced LC might be used for diagnosis and assessment of disease progression.
基金Supported by the Major State Basic Research Development Program of China (Program 973) (Grant No.2001CB510001)
文摘AIM:Gp96,also known as Grp94,is a member of heat shock protein (HSP) family and binds repertoires of peptides thereof eliciting peptide-specific T cell immune responses. It predominantly locates inside the endoplasmic reticulum (ER) with some cell surface expression in certain cancerous cells.Previous studies have shown that gp96 expression level was up-regulated in tumor cells,including hepatocellular carcinoma (HCC).However,relationship between the extent of gp96 expression and disease progression especially HBV-induced chronic infection,cirrhosis and hepatocellular carcinoma,has not been addressed before.As primary HCC can be induced and progressed from chronic hepatitis B virus (HBV) infection and HBV-induced cirrhosis,we designed an immunohistochemical experiment to test the correlation between gp96 expression level and HBV-induced disease progression,from chronic HBV infection,cirrhosis to HCC. METHODS:We chose liver samples from different patients of hepatitis B virus induced diseases,including chronic hepatitis B (77 patients),cirrhosis (27 patients) and primary HCC (30 patients),to test the expression level of gp96 in different affected groups.Formalin-fixed,and paraffin- embedded liver tissues taken from these patients were immuno-stained by using an anti-gp96 monoclonal antibody for the expression level of gp96 protein in the sections.In addition,Western blotting of whole cell lysates derived from established human embryonic liver cell lines and several human HCC cell lines (Huh7,HepG2,SSMC-7721) was compared with the expression of gp96. RESULTS:We found that the extent of elevated gp96 expression was significantly correlated with the disease progression,and was the highest in HCC patients,lowest in chronic HBV infection and was that of the cirrhosis in the middle. CONCLUSION:Increased expression of gp96 might be used as a diagnostic or prognostic bio-marker for the HBV infection and HBV-induced diseases.
基金Supported by the Major State Basic Research Development Program of China, Program 973, Grant No. 2001CB510001
文摘AIM: Most studies on the immune effect of gp96 were focused on its enhancement of CTLs. It is interesting to know whether gp96 could influence the humoral immune response, and whether the recombinant N-terminal fragment of gp96 could substitute native gp96 to stimulate the immune system.METHODS: gp96 isolated from livers of normal mice and its N-terminal fragment (amino acid 22-355) expressed in E coli were used for immunization of BALb/c mice. Eight groups of mice received one of the following regiments subcutaneously in 100 μL phosphate buffered saline (PBS)at an interval of 3 wk. Group 1: PBS only; group 2:gp96 only; group 3: N-terminal fragment only; group 4: HBsAg only; group 5: HBsAg+gp96; group 6: HBsAg+N-terminalfragment; group 7: HBsAg+incomplete Freud's adjuvant; group 8: HBsAg+N-terminal fragment (95 ℃ heated for 30 min). Serum anti-HBsAg antibody levels were assayed by ELISA. CTL responses in splenocytes were analyzed by ELISPOT after the last vaccination.RESULTS: The average titer of serum anti-HBsAg antibodyin the mice immunized with HBsAg together with gp96 or its N-terminal fragment were much higher than those immunized with HBsAg alone detected by ELISA. The cellular immune response of the mice immunized with HBsAg together with gp96 or its N-terminal fragment was not different with those immunized with HBsAg alone measured by ELISPOT assay.CONCLUSION: gp96 or its N-terminal fragment greatly improved humoral immune response induced by HBsAg, but failed to enhance the CTL response, which demonstrated the potential of using gp96 or its N-terminal fragment as a possible adjuvant to augment humoral immune response against HBV infection.
