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Multinucleated giant cell formation in BHK-21-528 cell monolayers infected with Japanese encephalitis viruses
Abstract:
Formation of prominent multinucleated giant cells (MGC) was observed in monolayers of a clonal line of BHK-21 cells (BHK-21-528) when infected with Japanese encephalitis virus (JEV). MGC were first observed 3 to 4 days after infection and cytopathic changes proceeded thereafter. Formation of MGC is a typical cytopathic change in this clonal cell line. Virus titer in 50% tissue culture infective dose (TCID50) equaled that in 50% MGC-forming dose. Virus titer in TCID50 was approximate to plaque-forming units (PFU) in the same host cells. An ability of JEV to form MGC was maintained at least for six serial passages in BHK-21-528. It was inactivated by heating at 56 C for 3 min. All JEV strains, except an attenuated live vaccine strain, induced formation of MGC in BHK-21-528 cells. Red blood cells of several animal species were not adsorbed to MGC induced by JEV. The MGC-forming ability of JEV was specifically neutralized by anti-JEV serum. By fluorescence antibody technique, the MGC were specifically stained by anti-JEV antibody conjugated with fluorescein isothiocyanate. Immunization of animals with lysates of the MGC resulted in production of antibodies against JEV, but no antibody against other viruses which have been reported to induce MGC formation. From these evidences, it was concluded that JEV induced formation of MGC in BHK-21-528.
Insights
Japanese encephalitis virus (JEV) infection of BHK-21 cells causes multinucleated giant cell (MGC) formation. This MGC-inducing ability is a characteristic feature of JEV, confirmed through various assays and antibody responses.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Japanese encephalitis virus (JEV) is a significant human pathogen.
- The cytopathic effects of JEV in cell culture are not fully characterized.
- Multinucleated giant cells (MGC) can be induced by various viral infections.
Purpose of the Study:
- To investigate the formation of multinucleated giant cells (MGC) in BHK-21 cells infected with Japanese encephalitis virus (JEV).
- To characterize the MGC-forming ability of JEV and its relationship to viral infectivity and antigenicity.
Main Methods:
- Infection of BHK-21 cell monolayers with JEV.
- Observation and quantification of MGC formation.
- Determination of virus titers (TCID50, PFU).
- Serial passage experiments.
- Heat inactivation assays.
- Neutralization assays with anti-JEV serum.
- Immunofluorescence staining.
- Animal immunization studies.
Main Results:
- JEV infection of BHK-21-528 cells induced prominent MGC formation, typically observed 3-4 days post-infection.
- The MGC-forming dose correlated with the 50% tissue culture infective dose (TCID50) and plaque-forming units (PFU).
- MGC formation ability was maintained through six serial passages and was heat-labile (inactivated at 56°C for 3 min).
- Most JEV strains induced MGC, except for an attenuated vaccine strain.
- MGC did not adsorb red blood cells from various animal species.
- Anti-JEV serum specifically neutralized JEV-induced MGC formation.
- Immunofluorescence confirmed JEV antigens within the MGC.
- Immunization with MGC lysates produced antibodies specific to JEV.
Conclusions:
- JEV induces the formation of multinucleated giant cells (MGC) in the BHK-21-528 cell line.
- MGC formation is a characteristic cytopathic effect of JEV infection.
- The MGC-forming ability is directly linked to JEV infectivity and antigenicity, serving as a potential marker for JEV activity.