Immunofluorescence studies on the replication of some arboviruses in nucleated and enucleated cells.
Porcine stable kidney (PS) or Vero cells infected with either flavi-(Japanese encephalitis--JE, West Nile--WN, and Dengue--DEN-2) or alphaviruses (Chikungunya--CHIK and Sindbis--SIN) were stained in indirect fluorescent antibody (FA) assay with anti-JE virus monoclonal (MoAb) Hx-3 (flavivirus cross-reactive) and polyclonal (immune PF) antibodies. By 48 hr post infection (p.i.), 15 to 20% of the three flaviviruses and CHIK virus infected cells, which revealed positive cytoplasmic immunofluorescence (IF), showed intranuclear IF. By 24 hr p.i., the intranuclear IF was not observed or became diminished. The enucleation of cells by cytochalasin B treatment prior to the infection with any of the three flaviviruses resulted in the loss of IF compared with the cells enucleated after the infection (18 hr p.i.) whereas SIN or CHIK virus-infected cells reacted similarly by the either method. These findings indicate an essential role of the nucleus in the replication of the flaviviruses only and while replicating in the infected cells, flaviviruses and CHIK virus might express viral specific proteins in the cell nuclei.
Lad
V J
VJ
National Institute of Virology, Pune, India.
Gupta
A K
AK
Ghosh
S N
SN
Banerjee
K
K
D000524:Alphavirus / Q000254:growth & development* / Q000302:isolation & purification; D000818:Animals; D001103:Arboviruses / Q000254:growth & development* / Q000302:isolation & purification; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000187:drug effects / Q000382:microbiology; D002478:Cells, Cultured / Q000187:drug effects; D003571:Cytochalasin B / Q000494:pharmacology; D003593:Cytoplasm / Q000382:microbiology; D018067:Flaviviridae / Q000254:growth & development* / Q000302:isolation & purification; D005455:Fluorescent Antibody Technique; D007668:Kidney / Q000166:cytology; D013552:Swine; D014709:Vero Cells / Q000187:drug effects
D003571:Cytochalasin B