Characteristics of escape mutations from occult hepatitis B virus infected patients with hematological malignancies in South Egypt

Figure 1 Phylogenetic tree constructed from the nucleotide sequences of the full hepatitis B virus genome. The phylogenetic tree is constructed by the neighbor joining method and significant bootstrap values (> 75%) are indicated in the tree roots. HBV sequences isolated from the studied cohort are indicated in bold. Reference sequences retrieved from the GenBank/EMBL/DDBJ are indicted by their accession numbers. The country origin of the reference sequences is indicated in brackets. HBV genotypes A-H are indicated in the cluster roots. HBV: Hepatitis B virus.

Figure 2 Expression of hepatitis B virus DNA and antigens after transfection of Huh 7 cells with wild type hepatitis B virus genotype D clone (WT) and mutants with S gene mutations P120T (120T) and S143L (143L). A and B represent the extracellular expression of HBsAg and HBcrAg, respectively, by WT and S gene mutants (120T and 143L); C represents the intracellular expression of the core gene product (HBcrAg) by WT and S gene mutant clones; D: The density of the single-stranded DNA in Southern blot analysis of cell lysates of Huh7 cells transfected with WT and mutants with S gene mutations.

Figure 3 Western blot analysis of hepatitis B surface antigen (A, C), hepatitis B virus core (B), core related proteins (D) expressed by hepatitis B virus genotype D clones (WT) and hepatitis B surface antigen mutant clones (120T and 143L) in the supernatant of Huh 7 cells. The results for the wild type clones in the three independent transfection experiments were similar and their mean value is set at 1.0. The hepatitis B surface antigen and hepatitis B virus core proteins levels are expressed relative to this value in (C) and (D). HBV: Hepatitis B virus.

Figure 4 Western blot analysis of hepatitis B surface antigen (A) and hepatitis B virus core (B) proteins expressed by hepatitis B virus genotype D clones (WT) and hepatitis B surface antigen mutant clones (120T and 143L) in the cell lysate of Huh 7 cells. HBV: hepatitis B virus.

Figure 5 Genotype distribution of the detected S gene product mutation among the database reference sequences. HBV: Hepatitis B virus.