Susan Tai

Combination of alpha-1-acid glycoprotein and alpha-fetoprotein as an improved diagnostic tool for hepatocellular carcinoma.

BACKGROUND To evaluate the diagnostic value of alpha-1-acid glycoprotein (AAG) and the combination with alpha-fetoprotein (AFP) in hepatocellular carcinoma (HCC) patients. METHODS AAG was measured in serum of 65 HCC patients and 54 chronic liver diseases (CLD) patients by using proteomic approach. Sensitivity and specificity of AAG and its combination with AFP were determined and compared with AFP alone for the diagnosis of HCC. RESULTS The expression concentration of AAG was significantly higher in HCC patients than chronic liver disease with sensitivity (77%) and accuracy (83%). Receiver operating characteristic analysis yielded the following AUC: AFP 0.750 (CI 95% 0.663-0.837), AAG 0.907 (CI 95% 0.855-0.960) and AFP+AAG 0.943 (CI 95% 0.897-0.988). At a specificity of 90%, the combination of AFP+AAG had sensitivity 89% and accuracy 90%, which was higher than sensitivity (52.3%) and accuracy (70%) when using AFP alone. CONCLUSION The combination of AAG and AFP shows high sensitivity and improves the accuracy of HCC diagnosis.

Genotype diversity of hepatitis C virus (HCV) in HCV-associated liver disease patients in Indonesia

Background: Hepatitis C virus (HCV) genotype distribution in Indonesia has been reported. However, the identification of HCV genotype was based on 5′‐UTR or NS5B sequence.

Hepatitis C virus genotype in blood donors and associated liver disease in Indonesia.

Objective: The aim of this study was to investigate the distribution of hepatitis C virus (HCV) genotype and the possible association between genotype and HCV-associated liver disease in Indonesia. Methods: 32 anti-HCV-positive asymptomatic carriers (AC), 55 chronic hepatitis (CH), 41 liver cirrhosis (LC), and 35 hepatocellular carcinoma (HCC) patients were included in this study. HCV genotyping was performed by phylogenetic analysis of the NS5B and 5′-UTR regions. Results: The HCV subtype 1b (36.5%), based on NS5B region, was the most prevalent, followed by subtypes 3k (15.4%), 2a (14.4%), 1a (12.5%) and 1c (12.5%), and 2e (4.8%). Subtypes 2f, 3a, 3b, and 4a were also found in some of the samples. HCV subtypes 3k (40.0%) and 1a (35.0%) were the two major subtypes in AC. HCV subtype 1b was not found in AC, but it was common in CH (31.3%), LC (50.0%), and HCC (57.1%). Conclusion: HCV subtype 1b was prevalent in samples of HCV-associated liver disease patients, including CH, LC and HCC. The percentage of subtype 1b was increased with the disease severity (AC < CH < LC < HCC).

Hepatitis B virus genotypes/subgenotypes in voluntary blood donors in Makassar, South Sulawesi, Indonesia

BackgroundHepatitis B virus (HBV) genotype appears to show varying geographic distribution. Molecular epidemiological study of HBV in particular areas in Indonesia is still limited. This study was aimed to identify the prevalence of HBV genotype/subgenotype and mutations in basal core promoter (BCP) region in voluntary blood donors in Makassar, one of the biggest cities in east part of Indonesia.A total of 214 hepatitis B surface antigen (HBsAg)-positive samples were enrolled in this study. HBV genotype/subgenotype was identified by genotype-specific PCR method or direct sequencing of pre-S region. Mutations in BCP were identified by direct sequencing of the corresponding region.ResultsHBV/B and HBV/C were detected in 61.21% and 25.23% of the samples, while mix of HBV/B and HBV/C was found in 12.62% of the samples. Based on pre-S region, among HBV/B and HBV/C, HBV/B3 (95.00%) and HBV/C1 (58.82%) were predominant. Interestingly, HBV/D was identified in two samples (22.165.07 and 22.252.07). Complete genome sequences of two HBV/D strains (22.165.07 and 22.252.07) demonstrated that both strains belong to HBV/D6, and the divergence between the two strains were 1.45%, while divergences of both 22.165.07 and 22.252.07 strains with reference strain (AM422939/France) were 2.67%. A1762T/G1764A mutation was observed in 1.96% and 5.36%, whereas T1753V mutation was found in 2.94% and 1.79% of HBV/B and HBV/C, respectively.ConclusionHBV/B and HBV/C are dominant in Makassar, similar to most areas in Indonesia. Mutations in BCP which might be associated with severity of liver disease are less common.

Alpha-1-acid glycoprotein as potential biomarker for alpha-fetoprotein-low hepatocellular carcinoma

BackgroundThe outcome of patients with hepatocellular carcinoma (HCC) remains poor because of late diagnosis. We determined the performances of α -1-acid glycoprotein (AAG) and des-γ-carboxy prothrombin (DCP) for the diagnosis of HCC, especially for α-fetoprotein (AFP)-low HCC.MethodsOf the 220 patients included in this retrospective study, 124 had HCC, and 61 (49%) of these were AFP-low HCC (AFP ≤ 20 ng/mL). The remaining 96 patients, including 49 with chronic hepatitis B or C and 47 with cirrhosis, were considered as control. Plasma AAG was analyzed using high performance liquid chromatography (HPLC) and confirmed using Western blot technique.ResultsWhen all patients with HCC were evaluated, the area under receiver operating characteristic (ROC) curves for AAG (0.94, 95% CI: 0.91-0.97) and DCP (0.92, 95% CI: 0.88-0.95) were similar (P = 0.40). AAG had better area under ROC curve (0.96, 95% CI: 0.94-0.99) than DCP (0.87, 95% CI: 0.81-0.93) for AFP-low HCC (P < 0.05). At the specificity 95%, the sensitivity of AAG was higher in AFP-low HCC than in AFP-high HCC (82% and 62%, respectively). In contrast, higher sensitivity was obtained from DCP in discriminating HCC patients with low AFP than that in high AFP (57% and 90%, respectively).ConclusionOur cross-sectional study showed that AAG was better performance in diagnosing HCC patients with low AFP, while DCP did better in those with high AFP.

Low prevalence of hepatitis B virus pre-S deletion mutation in Indonesia.

The molecular epidemiological study of hepatitis B virus (HBV) in Indonesia is still limited. This study was aimed to identify the prevalence of HBV pre‐S deletion/insertion mutations, and to assess the association of pre‐S deletion mutation with liver disease progression in Indonesia. Pre‐S mutations were identified by direct sequencing. Of the 265 subjects, 32 samples (12.1%) harbored pre‐S deletion/insertion mutations. The prevalence of those pre‐S mutations was 2.7% (2/75), 12.9% (8/62), 16.7% (11/66), and 17.7% (11/62) in asymptomatic carrier, chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma groups, respectively. Statistical analysis showed significant difference among them (P = 0.024). In HBV genotype B (HBV/B), pre‐S1, pre‐S1/S2, and pre‐S2 deletion mutations were detected respectively in 3 (17.6%), 4 (23.5%), and 9 (52.9%) of 17 samples. On the other hand, in HBV/C, 12 of 15 samples (80.0%) showed a pre‐S2 deletion mutation, and only 2 samples (13.3%) demonstrated a pre‐S1/S2 deletion mutation. These results suggest that in HBV/B deletion mutation tends to occur in pre‐S1 or pre‐S1/S2 region, while in HBV/C the deletion mutation usually occurs in the pre‐S2 region. Analysis of complete genome of four viruses confirmed that 3 isolates were classified into HBV/B3, and 1 isolate was HBV/C1. However, SimPlot and BootScan analyses showed that isolate 08.10.002 was an intragenotypic recombinant between HBV/B3 and HBV/B4. As conclusion, the prevalence of HBV pre‐S mutations was relatively low in Indonesian patients compared to those from Taiwan, Japan, and other Asian countries. There was a weak association between pre‐S deletion mutation and progressive liver disease. J. Med. Virol. 83:1717–1726, 2011.

Association of core promoter mutations of hepatitis B virus and viral load is different in HBeAg(+) and HBeAg(-) patients

AIM To identify the prevalence of hepatitis B e antigen (HBeAg) and to assess the association of hepatitis B virus (HBV) core promoter mutations and viral load in Indonesian patients. METHODS Sixty-four patients with chronic hepatitis, 65 with liver cirrhosis and 50 with hepatocellular carcinoma were included in this study. HBeAg and hepatitis B e antibody (HBeAb) tests were performed using enzyme-linked immunosorbent assay and the mutations were analyzed by sequencing. Viral load was measured by real-time polymerase chain reaction. RESULTS Of 179 patients, 108 (60.3%) were HBeAg(-) and 86 (79.6%) of these HBeAg(-) patients had been seroconverted. The A1896 mutation was not found in HBeAg(+) patients, however, this mutation was detected in 70.7% of HBeAg(-) patients. This mutation was frequently found when HBeAg was not expressed (87.7%), compared to that found in HBeAg seroconverted patients (65.1%). The A1899 mutation was also more prevalent in HBeAg(-) than in HBeAg(+) patients (P = 0.004). The T1762/A1764 mutation was frequently found in both HBeAg(+) and HBeAg(-) patients, however, the prevalence of this mutation did not significantly differ among the two groups (P = 0.054). In HBeAg(+) patients, the T1762/A1764 mutation was correlated with lower HBV DNA (P < 0.001). The A1899 mutation did not correlate with HBV DNA (P = 0.609). In HBeAg(-) patients, the T1762/A1764 mutation alone was not correlated with HBV DNA (P = 0.095), however, the presence of either the T1762/A1764 or A1896 mutations was associated with increased HBV DNA (P < 0.001). CONCLUSION The percentage of HBeAg(-) patients is high in Indonesia, and most of the HBeAg(-) patients had been seroconverted. The A1896 mutation was most likely the major cause of HBeAg loss. The T1762/A1764 mutation alone was associated with lower viral loads in HBeAg(+) patients, but not in HBeAg(-) patients.

Immunoregulatory effects of AFP domains on monocyte-derived dendritic cell function

BackgroundAlpha-fetoprotein (AFP) is a tumor-associated glycoprotein that functions in regulation of both ontogenic and oncogenic growth. Recent study showed that AFP can induce apoptosis or impair monocyte-derived dendritic cell (MDDC) function. However, it is still unclear which AFP domain (D-AFP) plays major role in this function.ResultsAs expected monocytes cultured in the presence of Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF) and Interleukin-4 (IL-4) developed into MDDC. Up-regulation of HLA-DR and CD11c as well as loss of CD14 molecules could be observed. Full length AFP (FL-AFP), domain 2 AFP (D2-AFP) and D3-AFP, but not D1-AFP, significantly inhibited the expression of HLA-DRhigh/CD11chigh and CD80+/CD86high molecules. In contrast, CD83 expression was substantially down-regulated in all samples. Expression of CD40 was significantly suppressed by FL-AFP but not by any D-AFPs. Finally, both FL-AFP and D-AFP impaired the MDDC ability to secrete IL-12 (p70).ConclusionsD2- and D3- but not D1-AFP extensively suppresses the MDDC function. All the recombinant AFP proteins impaired the ability of MDDC to secrete IL-12.

Hepatitis B Virus Double Mutations is There any Role in Pathogenesis of Hepatocellular Carcinoma in Young Patients

Background: The incidence of hepatocellular carcinoma (HCC) below age 40 years old in our institution were relatively high compared with other institutions in Asia. Hepatitis B virus (HBV) basal core promoter (BCP) double mutations correspond with increasing age. The aim of this study was to know if there was any role of HBV double mutations in young HCC patients. Method: A descriptive study was performed on HBV related HCC patients in Cipto Mangunkusumo Hospital in May 2006-November 2008. Patient were recruited consecutively and divided in to two groups, below 40 and above 40 years old. The genotypes were examined by polymerase chain reaction (PCR) method. The alpha feto protein (AFP) values were diagnosed based on ELISA method. The BCP A1762T/G1764A double mutations were examined by direct sequencing. Results: There were 49 HBV related HCC samples consist of 14 (28.5%) samples with age below 40 years old and 35 (71.5%) samples with age above 40 years old. We only found two genotype, genotype B was dominant in patients with HBV related HCC compare to genotype C, 43 (88%) and 6 (12%) respectively. The increasing of AFP level above 400 ng/mL was only found in about half of the samples, 7 (50%) 40 years old. Double mutations of A1762T/G1764A in BCP occurred in 5 (36%) 40 years old. Conclusion: The incidence of HBV related HCC in young patients were relatively high. The proportion of patients with AFP level < 400 ng/mL in patients below 40 years old were higher compared to patients above 40 years. Keywords : hepatocellular carcinoma, BCP double mutation, HBV genotype