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Selection and characterization of an interferon-responsive clonal cell line of HeLa cells
Abstract:
HeLa cells generally do not respond well to interferon (IFN). We have used is-1, an IFN-sensitive mutant of mengovirus, to select a clone of IFN-responsive HeLa cells (F-H12). At moderate levels of human alpha/beta IFN, is-1 yields were fivefold lower in these cells than in similarly protected control cells. In contrast, wild-type mengovirus, vesicular stomatitis virus and a wild-type and thymidine kinase-negative strains of herpes simplex virus type 1 grew equally well in both cell lines. By a cell survival assay, the F-H12 line was up to 100 times more responsive to IFN than the parental line when challenged by is-1. 2'-5'-Oligo(A)-dependent endonuclease activity was the same in both lines. These observations cannot be accounted for by enhanced induction of IFN following infection.
Insights
Researchers developed an interferon-responsive HeLa cell line (F-H12) by using an interferon-sensitive mengovirus mutant. This new cell line shows significantly enhanced antiviral responses to interferon, particularly against the specific mutant virus.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- HeLa cells typically exhibit poor responsiveness to interferon (IFN).
- Interferon is a crucial component of the innate immune system, mediating antiviral states in cells.
- Developing cell models with enhanced IFN responses is vital for studying viral infections and antiviral mechanisms.
Purpose of the Study:
- To select and characterize a clone of HeLa cells with enhanced sensitivity to interferon.
- To investigate the antiviral efficacy of the selected cell line against different viruses.
Main Methods:
- Utilized an interferon-sensitive mengovirus mutant (is-1) to screen and select for interferon-responsive HeLa cells.
- Compared the growth of is-1, wild-type mengovirus, vesicular stomatitis virus, and herpes simplex virus type 1 in parental and selected cell lines.
- Assessed cell survival and 2'-5'-Oligo(A)-dependent endonuclease activity.
Main Results:
- A clone of interferon-responsive HeLa cells (F-H12) was successfully selected.
- F-H12 cells demonstrated a fivefold reduction in is-1 yields compared to control cells at moderate interferon levels.
- F-H12 cells exhibited up to 100-fold greater responsiveness to interferon when challenged with is-1, as measured by cell survival.
- Growth of other tested viruses (wild-type mengovirus, VSV, HSV-1) was not significantly affected in F-H12 cells.
- No difference in 2'-5'-Oligo(A)-dependent endonuclease activity was observed between the cell lines.
Conclusions:
- The F-H12 cell line represents a valuable tool for studying interferon-mediated antiviral responses, particularly against specific viral mutants.
- The enhanced interferon responsiveness in F-H12 cells is specific and not due to general increases in interferon-induced antiviral pathways like endonuclease activity.
- The selection strategy using an IFN-sensitive mutant is effective for generating IFN-responsive cell lines.