Functional analysis of a rare HBV deletion mutant in chronically infected children.
Liver damage caused by chronic hepatitis B virus (HBV) infection may be enhanced through the selection of deleted HBV preS mutants by intracellular accumulation of viral proteins and subsequent cell death. However, the prevalence and impact of such mutants on the clinical course of infection have not yet been studied in children. Serum samples from 60 children (mean age 9.8 y) were investigated by means of PCR and direct sequencing of the entire preS region. Only one patient (1.5%) was found with a mixed HBV population of a deletion spanning 183 nucleotides and wild-type sequences. This mutation alters the HBV large-surface protein and removes the small-surface promoter. To clarify the significance of this mutation, we studied 14 serial serum samples of the child within a follow-up of 10 y. After occurrence of the mutation, the liver enzymes increased, despite seroconversion to anti-HBe. Transfection of an HBV expression construct containing this deletion into human hepatoma cells by using an HBV in vitro replication system showed that the mutant lost the ability of nucleocapsid packaging as a result of alteration of the transmembrane topology of the large surface protein. This effect could not be restored by coexpression of wild-type large- or small-surface proteins in trans. In conclusion, the circulation of HBV preS deletion mutants is rare in childhood. However, our functional and clinical follow-up studies in one child suggest that such a mutant may have the potential to aggravate liver inflammation, especially if corroborated with larger numbers of children.
Gerner
Patrick
P
Children's Hospital, Helios Klini Kum, Wuppertal, Heusnerstrasse 40, D-42283 Wuppertal, Germany.
Schäfer
Heike Marie Clara
HM
Prange
Reinhild
R
Pravitt
Dirk
D
Wirth
Stefan
S
D000293:Adolescent; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005260:Female; D006515:Hepatitis B virus / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000235:genetics*; D006801:Humans; D007223:Infant; D008297:Male; D016133:Polymerase Chain Reaction; D017384:Sequence Deletion
D017931:DNA Primers