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Ultrastructural Studies of Fibroblasts Transfected with Hepatitis B Virus DNA
Author(s) -
Aoki Naoto,
Gerber Michael A.,
Thung Swan N.,
Chen MeiLing,
Christman Judith K.,
Price Peter M.,
Flordellis Christos S.,
Acs George
Publication year - 1984
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840040115
Subject(s) - transfection , immunoelectron microscopy , hbsag , endoplasmic reticulum , microbiology and biotechnology , biology , hepatitis b virus , virology , virus , cell culture , antibody , immunology , genetics
Cultured 3T3 mouse fibroblasts transfected with cloned hepatitis B virus genome and DNA coding for methotrexate‐resistant dihydrofolate reductase, produce and secrete significant amounts of hepatitis B surface antigen (HBsAg). Ultrastructural morphometry revealed that fibroblasts transfected with hepatitis B virus DNA contained significantly more lysosomes than did fibroblasts transfected with the gene coding for methotrexate resistance or normal fibroblasts. Although abundant HBsAg was found in the cytoplasm of transfected fibroblasts by immunologic methods, HBsAg particles were not detected by electron microscopy. Immunoelectron microscopy localized HBsAg to the nuclear envelope, rough endoplasmic reticulum, and endoplasmic cisternae. These findings suggest that the transfected cells produce mainly nonparticulate HBsAg or that they have a defect in intracisternal packaging of HBsAg into particles.