J Korean Med Sci.  2005 Jun;20(3):433-437. 10.3346/jkms.2005.20.3.433.

Overlapping Gene Mutations of Hepatitis B Virus in a Chronic Hepatitis B Patient with Hepatitis B Surface Antigen Loss during Lamivudine Therapy

• Lee SY
• Choi MS
• Lee D
• Lee JH
• Koh KC
• Paik SW
• Yoo BC

• KMID: 1778502
• DOI: http://doi.org/10.3346/jkms.2005.20.3.433

Abstract

Disappearance of hepatitis B surface antigens (HBsAg) in chronic hepatitis B usually indicates clearance of hepatitis B virus (HBV) infection. However, false HBsAg negativity with mutations in pre-S2 and 'a' determinant has been reported. It is also known that YMDD mutations decrease the production of HBV and escape detection of serum HBsAg. Here, we report overlapping gene mutations in a patient with HBsAg loss during the lamivudine therapy. After 36 months of lamivudine therapy in a 44-yrold Korean chronic hepatitis B patient, serum HBsAg turned negative while HBV DNA remained positive by a DNA probe method. Nucleotide sequence of serum HBV DNA was compared with the HBV genotype C subtype adr registered in NCBI AF 286594. Deletion of nucleotides 23 to 55 (amino acids 12 to 22) was identified in the pre-S2 region. Sequencing of the 'a' determinant revealed amino acid substitutions as I126S, T131N, M133T, and S136Y. Methionine of rtM204 in the P gene was substituted for isoleucine indicating YIDD mutation (rtM204I). We identified a HBV mutant composed of pre-S2 deletions and 'a' determinant substitutions with YMDD mutation. Our result suggests that false HBsAg negativity can be induced by combination of overlapping gene mutations during the lamivudine therapy.

Keyword

Hepatitis B, Chronic; Hepatitis B Surface Antigens; Hepatitis B virus; Lamivudine; Mutation

MeSH Terms

Adult
Amino Acid Sequence
Anti-HIV Agents/therapeutic use
Base Sequence
Comparative Study
DNA Mutational Analysis
DNA, Viral/blood/chemistry/genetics
DNA-Directed DNA Polymerase/genetics
Gene Deletion
Genes, Overlapping/*genetics
Hepatitis B Surface Antigens/blood/*genetics
Hepatitis B virus/*genetics
Hepatitis B, Chronic/blood/*drug therapy/virology
Humans
Lamivudine/*therapeutic use
Male
Molecular Sequence Data
*Mutation
Protein Precursors/genetics
Sequence Alignment
Sequence Homology, Amino Acid
Sequence Homology, Nucleic Acid
Viral Proteins/genetics

Figure

• 

  Fig. 1 Alignment of the sequence with HBV subtype adr in the pre-S2 region. Comparisons of nucleotide sequences in the S region between mutant HBV and wild type HBV (NCBI nucleotide LOCUS AF286594) reveal the deletion of nucleotide 23 to 55 in pre-S2 gene.

• 

  Fig. 2 Translation of the pre-S2 region open reading frame aligned with the wild type. The heterogeneous amplication products reveal the presence of pre-S2 deletions. An arrow indicates pre-S2 deletions which is consistent with deletion of amino acids 12 to 22 in the pre-S2 region. Small boxes indicate the locations of amino acid substitutions within the 'a' determinant.

• 

  Fig. 3 Amino acid substitution within the 'a' loop structure of HBsAg protein. Sequencing revealed substitutions at position I126S, T131N, M133T, and S136Y in the 'a' determinant of the S gene. Black circles represent residues with mutations while gray circles represent cysteines. White circles represent other residues.

• 

  Fig. 4 Nucleotide and amino acid sequences of the polymerase gene. Methionine on rtM204 is substituted by isoleucine indicating YIDD mutation (rtM204I).

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