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Vet Microbiol ; 86(1-2): 165-74, 2002 Apr 22.
Artículo en Inglés | MEDLINE | ID: mdl-11888699

RESUMEN

Herpes simplex virus-1 (HSV-1) based amplicon vectors are promising gene delivery vehicles because they have a large transgene capacity and can efficiently transduce many different cell types, including non-dividing cells, of various animal species. The Circ protein of bovine herpesvirus-1 (BHV-1) is a myristylated virion component of unknown function. Preliminary experiments with a circ gene deletion mutant indicated that Circ may influence the host's immune response by downregulating MHC-II expression in bovine monocytes. To get more insight into the function of Circ, amplicon vectors were constructed with various open reading frames (ORFs) under the control of the HSV-1 IE4/5 promoter: (i) the Circ ORF alone, (ii) a fusion ORF encoding an N-terminal Circ fused to the enhanced green fluorescent protein (eGFP), (iii) the eGFP ORF alone, and (iv) the Circ ORF in the inverted orientation. Upon helpervirus-free packaging into HSV-1 amplicon particles and transduction of Vero cells, both Circ alone and the Circ-eGFP fusion protein produced a punctate pattern within the cytoplasm, suggesting membrane association of the myristylated protein. In contrast, eGFP alone was evenly distributed over the cytoplasm of transduced cells. Upon infection of bovine buffy-coat cells, it was observed that cells of the monocyte lineage but not lymphocytes were transduced. Transgene expression reached a peak around 20h after transduction and lasted for at least 90h. Transduced monocytes underwent specific morphological changes, which may be attributed to Circ synthesis.


Asunto(s)
Terapia Genética/veterinaria , Herpesvirus Bovino 1/genética , Proteínas Virales/genética , Animales , Bovinos , Chlorocebus aethiops , Efecto Citopatogénico Viral/genética , Citometría de Flujo/veterinaria , Terapia Genética/métodos , Vectores Genéticos , Proteínas Fluorescentes Verdes , Herpesvirus Bovino 1/química , Herpesvirus Humano 1/química , Herpesvirus Humano 1/genética , Proteínas Luminiscentes/química , Monocitos , Plásmidos/genética , Transducción Genética/métodos , Células Vero , Proteínas Virales/química
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