Related Experiment Videos
Infection and transformation of mouse cells by human adenovirus type 2
Abstract:
The susceptibilities of C57Bl/6J mouse embryo fibroblasts (MEF) and baby mouse kidney (BMK) cells to infection or transformation by adenovirus type 2 have been compared to those of rat embryo fibroblasts (REF). Both MEF and BMK cells were some 10-fold less permissive to replication of the virus than were REF cells, even though similar fractions of all three cell types, a maximum of 50-60%, produced viral tumor and structural antigens. This observation suggests that a very late step in adenovirus production, such as assembly or maturation, occurs much less efficiently in mouse cells than it does in rat cells. No significant differences in the frequencies of transformation, as assayed by the appearance of foci of morphologically transformed cells, were observed following transfection of adenovirus type 2 or type 5 DNA into the three cell types. However, it proved extremely difficult to establish permanent lines of adenovirus-transformed mouse cells: only 2 of more than 100 attempts were successful, compared to a success rate of close to 100% with adenovirus type 2-transformed REF or SV40-transformed MEF or BMK cells. The two lines of type 2 adenovirus-transformed MEF that were established have been shown to retain and express viral genetic information.
Insights
Mouse cells are less efficient at replicating adenovirus type 2 than rat cells, suggesting a late-stage defect in virus production. Establishing adenovirus-transformed mouse cell lines proved exceptionally difficult.
Area of Science:
- Virology
- Cell Biology
- Molecular Biology
Background:
- Adenovirus type 2 infects various cell types, including rodent fibroblasts.
- Susceptibility to viral infection and transformation can vary significantly between cell lines and species.
Purpose of the Study:
- To compare the permissiveness of mouse embryo fibroblasts (MEF) and baby mouse kidney (BMK) cells to adenovirus type 2 infection and transformation versus rat embryo fibroblasts (REF).
- To investigate potential differences in adenovirus replication, antigen production, and transformation efficiency between mouse and rat cells.
Main Methods:
- Infection of MEF, BMK, and REF cells with adenovirus type 2.
- Assay of viral replication and production of viral tumor and structural antigens.
- Transfection of adenovirus type 2 or 5 DNA into cells to assess transformation frequencies.
- Attempting to establish permanent adenovirus-transformed cell lines.
Main Results:
- MEF and BMK cells were approximately 10-fold less permissive to adenovirus type 2 replication than REF cells.
- Similar percentages of all cell types produced viral antigens, indicating a late-stage defect in mouse cells.
- Transformation frequencies were comparable across cell types, but establishing permanent transformed mouse cell lines was highly challenging (2/100+ attempts).
- Established transformed MEF lines retained and expressed viral genetic information.
Conclusions:
- Mouse cells exhibit a defect in a late stage of adenovirus type 2 replication, likely assembly or maturation.
- While adenovirus can transform mouse cells, establishing permanent transformed lines is inefficient compared to rat cells.
- Further research is needed to elucidate the specific molecular mechanisms underlying these species-specific differences in adenovirus replication and transformation.