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Demonstration of interference between dengue virus types in cultured mosquito cells using monoclonal antibody probes
Abstract:
Cultured Aedes albopictus cells (clone C6/36), persistently infected (PI) with dengue virus type 1 (dengue-1) were found resistant to superinfection with dengue virus type 3 (dengue-3). This was determined by indirect immunofluorescent (IF) staining of cultures using monoclonal antibody against a dengue-3 type-specific antigen. Dengue-1 PI cultures stained with this antibody 3 days after superinfection with dengue-3 virus (m.o.i of 2) had dengue-3 antigen in 0.1 to 1.0% of the cells. Control cultures infected with dengue-3 at the same multiplicity contained dengue-3 antigen in greater than 90% of the cells. The resistance to superinfection was not interferon-mediated, and occurred within 20 h after primary infection. In cultures simultaneously infected with two dengue virus types, one virus type was excluded from replication in most cells. A small population of cells was also found (about 1%) that contained type-specific antigen of both dengue virus types.
Insights
Aedes albopictus cells persistently infected with dengue virus type 1 resist superinfection by dengue virus type 3. This resistance, observed within 20 hours, is not interferon-mediated, impacting viral replication.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Persistent viral infections can alter host cell susceptibility to secondary infections.
- Understanding mechanisms of viral interference is crucial for controlling arbovirus transmission.
Purpose of the Study:
- To investigate the phenomenon of superinfection exclusion in dengue virus-infected Aedes albopictus cells.
- To determine the kinetics and underlying mechanisms of this resistance.
Main Methods:
- Cultured Aedes albopictus (C6/36) cells persistently infected with dengue virus type 1 (DENV-1).
- Superinfection with dengue virus type 3 (DENV-3) at a multiplicity of infection (m.o.i.) of 2.
- Indirect immunofluorescence (IF) staining using DENV-3 type-specific monoclonal antibodies.
- Assessment of viral antigen presence in cells at different time points.
Main Results:
- DENV-1 persistently infected cells showed significant resistance to DENV-3 superinfection, with only 0.1-1.0% of cells positive for DENV-3 antigen compared to >90% in control cells.
- Resistance to superinfection was established within 20 hours post-primary infection.
- The observed resistance was independent of interferon-mediated mechanisms.
- In co-infected cultures, one dengue virus type excluded the replication of the other in the majority of cells.
Conclusions:
- Persistent dengue virus type 1 infection induces a robust superinfection exclusion mechanism in Aedes albopictus cells.
- This phenomenon limits the co-replication of different dengue virus serotypes within individual mosquito cells.
- The rapid onset and non-interferon nature of this resistance suggest intrinsic cellular defense pathways are involved.