Detection of the hepatitis B virus X protein (HBx) antigen and anti-HBx antibodies in cases of human hepatocellular carcinoma.
Hepatitis B virus X protein (HBx) expressed in Escherichia coli DH5alpha by recombinant DNA technology was purified to homogeneity by use of glutathione-Sepharose beads. Immunological characterization of the recombinant HBx protein was performed. Specific binding between the anti-HBx monoclonal antibody and HBx protein showed the specificity of the recombinant HBx protein. The intact HBx protein of the factor Xa-digested glutathione S-transferase-HBx fusion protein was further purified and was used as an antigen for screening the titers of anti-HBx antibodies in sera. Titers of anti-HBx in sera from 20 patients with hepatocellular carcinoma (HCC), 20 patients with chronic hepatitis (CH), and 20 healthy individuals were evaluated by Western blotting and a quantitative enzyme-linked immunosorbent assay. The results indicated that 70% of sera from HCC patients and 5% of sera from CH patients contained antibodies with significant binding to the HBx protein. Western blotting of HBx protein in liver extracts from 20 HCC patients was also performed by using the anti-HBx monoclonal antibody. Results showed that 85% of HCC patients' liver tissues contained a specific HBx protein with the same molecular size as the purified intact HBx. Full correlation was found between anti-HBx antibody positivity in serum and HBx protein positivity in HCC tissues. The data demonstrated that the etiology of HCC is involved with hepatitis B virus (HBV) infection and that HBx in particular plays a role in the development of HBV-related HCC.
Hwang
Guang-Yuh
GY
Department of Biology, Life Science Research Center, Taichung Veterans General Hospital, Taichung, Taiwan, Republic of China. gyhwang@mail.thu.edu.tw
Lin
Chuang-Yu
CY
Huang
Li-Mei
LM
Wang
Yan-Hsiung
YH
Wang
Jing-Chyi
JC
Hsu
Ching-Ting
CT
Yang
Sheng-Shun
SS
Wu
Cheng-Chung
CC
D000328:Adult; D000368:Aged; D000914:Antibodies, Viral / Q000032:analysis / Q000097:blood*; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000276:immunology / Q000821:virology*; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D008113:Liver Neoplasms / Q000276:immunology / Q000821:virology*; D008297:Male; D008875:Middle Aged; D010957:Plasmids; D011994:Recombinant Proteins / Q000276:immunology; D015534:Trans-Activators / Q000097:blood / Q000276:immunology* / Q000302:isolation & purification*; D054334:Viral Regulatory and Accessory Proteins
D000914:Antibodies, Viral; D004279:DNA, Viral; D011994:Recombinant Proteins; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein