Superoxide anion production by granulocytes incubated with HeLa 229 cells persistently infected with influenza virus B/Lee/40.
OBJECTIVE
The purpose of this study was to determine whether granulocytes can be primed to produce superoxide anion by incubation with cells persistently infected with influenza viruses.


METHODS
The effect of in vitro incubation with HeLa 229 cells persistently infected with influenza virus B/Lee/40 (He/Le cells) on the generation of superoxide anion by human granulocytes was examined by a cytochrome c reduction assay.


RESULTS
Generation of superoxide anion from granulocytes, preincubated with He/Le cells in the productive phase and stimulated with formyl-methionine-leucine-phenylalanine, was significantly enhanced compared with granulocytes preincubated with uninfected HeLa 229 cells. He/Le cells in the productive phase were positive for viral proteins by immunofluorescence and for viral particles by scanning electron microscopy. The priming effects were concentration-dependently inhibited by polyclonal antibodies to the virus. He/Le cells in the latent phase, negative for viral proteins and viral particles, did not prime granulocytes. Culture supernatants of the productive phase did not prime granulocytes.


CONCLUSIONS
Unreleased influenza viruses on He/Le cells in the productive phase primed granulocytes to produce excessive amounts of superoxide anion, suggesting that cytotoxicity by hyperresponsive granulocytes could take place in persistent influenza infection as well as in acute infection.
Hayase
Yukiharu
Y
Department of Virology, Jichi Medical School, Tochigi, Japan. yuhayase@jichi.ac.jp
Tobita
Kiyotake
K
D000914:Antibodies, Viral; D003574:Cytochrome c Group / Q000378:metabolism; D006098:Granulocytes / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D009981:Influenza B virus / Q000502:physiology*; D010084:Oxidation-Reduction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013481:Superoxides / Q000378:metabolism*
D000914:Antibodies, Viral; D003574:Cytochrome c Group; D013481:Superoxides