Hepatitis B Virus Genotype B and High Expression of Interferon Alpha Receptor ß Subunit are Associated With Better Response to Pegylated Interferon Alpha 2a in Chinese Patients With Chronic Hepatitis B Infection.

BACKGROUND: Hepatitis B virus (HBV) is one of leading causes of various hepatic diseases including acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Hundreds of million people worldwide are infected by HBV, chronically. OBJECTIVES: This study in conducted to investigate the influence of Hepatitis B virus (HBV) genotypes and type I IFN-αreceptor ß subunit (IFNAR2) expression in liver on response to treatment with pegylated IFN-α-2a (Peg-IFN-α-2a) for chronic hepatitis B infection. PATIENTS AND METHODS: In this study, 65 eligible patients with chronic hepatitis B disease were enrolled. HBV genotypes of these patients were analyzed by using PCR-RFLP of the surface gene of HBV. The expression of IFNAR2 in the liver was immune histochemically investigated using anti-IFNAR2 antibody. All immune histochemical slides were read semi-quantitatively by image analysis. Chronic hepatitis B patients were treated with Peg-IFN-α2a therapy for a 48-week period and followed up for 24 weeks. Baseline characteristics and sustained viral response (SVR) to Peg-IFN-α-2a therapy were evaluated. RESULTS: 55 % of patients exhibited HBV genotype B and 31.7 % patients exhibited HBV genotypes C infections. After treatment with Peg-IFN-α-2a, SVR was achieved in 66.7 % of patients with HBV genotype B and in 26.3 % of patients with HBV genotype C (P = 0.009). Semiquantitative and the image analysis indicated by gray level values revealed a higher IFNAR2 expression in the group with severe inflammation (P < 0.001). Patients' high IFNAR2 protein expression had a significant impact on SVR to Peg-IFN-α-2a therapy (P = 0.028). CONCLUSIONS: HBV genotype B and high expression of IFNAR2 in the liver of chronic hepatitis B patients are closely associated with better response to Peg-IFN-α-2a therapy in chronic hepatitis B disease.

[Association of HBeAg positivity and alpha fetoprotein level with the prognosis of chronic severe hepatitis B].

OBJECTIVE: To explore the association of HBeAg positivity and alpha fetoprotein (AFP) level with the prognosis of chronic severe hepatitis B. METHODS: A total of 325 hospitalized patients with chronic severe hepatitis B were analyzed for serum HBeAg positivity and AFP levels and their association with the prognosis. RESULTS: Of all the 325 patients, 168 (51.7%) were HBeAg-positive and 157 (48.3%) were HBeAg-negative, and the two groups showed no significant difference in gender distribution, average peak value of total bilirubin and average trough prothrombin activity. Compared with the positive patients, the HBeAg-negative patients had significantly older age (P < 0.001), higher rate of liver cirrhosis (P < 0.001) and lower response rate (P < 0.05). Elevated AFP level was positively correlated to the response rate in both the HBeAg-positive (P < 0.001) and negative patients (P < 0.05). CONCLUSIONS: HBeAg-negative patients with chronic severe hepatitis B have poorer prognosis than the HBeAg-positive patients, and higher AFP levels are associated with more favorable prognosis regardless of the HBeAg positivity.

[Construction and expression of anti-tumor necrosis factor related apoptosis-inducing ligand receptor death receptor 5 chimeric antibody in eukaryotic cells].

OBJECTIVE: To construct the human/mouse chimeric antibody of a functional anti-death receptor 5 (DR5) antibody. Methods The viable region of light chain (VL) and viable region of heavy chain (VH) genes of anti-DR5 antibody were amplified and cloned into the light- and heavy-chain expression vectors respectively, then the recombinant plasmids were co-transfected into dihydrofolate reductase(-) Chinese hamster ovary cell (CHO-dhfr(-)) for expression. The positive clone was screened by the two selective genes (neo and dhfr). The humanization and specificity of chimeric antibody was identified by ELISA and Western blotting, and the tumoricidal activity of the expressed chimeric antibody was detected by tetrazolium salt phenazine methosulfate assay. RESULTS: The expression vectors stably expressed chimeric antibody in CHO-dhfr(-). In the cell supernatant of the F4' clone, the human IgG heavy constant region and light constant region were identified. Moreover, the secreted chimeric antibody retained the binding capacity to the antigen (DR5) and decreased the cell viability of Jurkat and HCT116 cells to 73.15% and 77.30% in vitro respectively. CONCLUSION: The human/mouse anti-DR5 chimeric antibody has been successfully expressed in eukaryotic cells and shows tumoricidal activity, which establishes a foundation for the future research of humanized antibody medicine.

Determination and analysis of complete nucleotide sequence of Chinese genotype D hepatitis B virus: the first report.

OBJECTIVE: To determine the complete nucleotide sequence of a Chinese hepatitis B virus (HBV) strain of genotype D. METHOD: The complete nucleotide sequence of HBV derived from a Chinese chronic asymptomatic carrier was amplified by PCR and cloned to conduct sequence analysis. Homology of the resulted nucleotide sequence with those of published HBV strains was assessed by using DNASIS, and complete sequence analysis of the phylogenetic tree of 30 genotype D HBV strains conducted by the assistance of Clustalw. RESULTS: With the complete nucleotide sequnce of 3 182 bp, this HBV strain belongs to ayw3 subtype and D genotype, with the Genebank accession number AF280817. Its complete nucleotide homology is 98.3% to 94.5% with the published sequences of genotype D HBV strains and less than 89.5% with the other HBV strains of genotype A, B, C, E, F and G published in GenBank. CONCLUSION: The evolutionary relations of the complete nucleotide sequence of this HBV strain is the closest to those of the 4 Swedish strains out of the 30 genotype D strains published in GenBank.