Enhanced green fluorescent protein as a marker for localizing murine cytomegalovirus in acute and latent infection.
A recombinant murine cytomegalovirus (mCMV) that expresses enhanced green fluorescent protein (EGFP) under control of the native immediate-early 1/3 promoter was constructed to detect directly sites of viral activity in latent and reactivated infections. The recombinant virus had acute and latent infection characteristics similar to those of wild-type mCMV. Rare green-fluorescing foci were observed in paraffin sections from lungs and spleens infected latently. Positive immunoperoxidase staining for EGFP in sections of the same lung tissues suggests that these cells may be sites of restricted viral gene expression. EGFP was detected easily in tissue explants reactivating from latent infection in vitro. Morphology and adhesion characteristics of fluorescing cells suggest that viral reactivation occurs in tissue macrophages in explant cultures. The observations presented in this study demonstrate the usefulness of EGFP-expressing recombinants as tools for direct tracking of mCMV activity in vivo and in vitro.
Henry
S C
SC
Department of Veterans Affairs, Medical Research Service and the Research Center on AIDS and HIV Infection, 111-H, 508 Fulton Street, Durham, NC 27705, USA.
Schmader
K
K
Brown
T T
TT
Miller
S E
SE
Howell
D N
DN
Daley
G G
GG
Hamilton
J D
JD
D000208:Acute Disease; D000818:Animals; D015415:Biomarkers; D015152:Blotting, Northern; D015139:Blotting, Southern; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D016650:Fluorescein-5-isothiocyanate; D049452:Green Fluorescent Proteins; D006566:Herpesviridae Infections / Q000821:virology*; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D018146:Muromegalovirus / Q000235:genetics / Q000254:growth & development* / Q000648:ultrastructure; D011401:Promoter Regions, Genetic; D014775:Virus Activation; D017735:Virus Latency
D015415:Biomarkers; D004274:DNA, Recombinant; D004279:DNA, Viral; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D016650:Fluorescein-5-isothiocyanate