Cell type-specific expression in brain cell cultures from a short human cytomegalovirus major immediate early promoter depends on whether it is inserted into herpesvirus or adenovirus vectors.

Abstract

Expression from a short human cytomegalovirus (HCMV) major immediate early (IE) promoterenhancer was tested in three different virus vectors : recombinant adenovirus (Ad), recombinant herpes simplex virus type 1 (HSV-1) and HSV-1-derived amplicon vectors. The HCMV major IE promoterenhancer within a replication-deficient recombinant Ad vector was shown to produce cell-specific expression in rat nervous system cell cultures. Recombinant Ad entered all cell types examined but the HCMV major IE promoter was silent in primary cultures of neocortical neurons and Schwann cells, although it drove transgene expression in astrocytes and fibroblasts. Moreover, in neurons and Schwann cells, expression from the HCMV major IE

Effects of ectopic decorin in modulating intracranial glioma progression in vivo, in a rat syngeneic model

“…Construction of the RAd/hCMV/b-gal and RAd/hCMV/ HSV1-TK has been described in detail elsewhere. 21,40 To create the shuttle vector pDE1/GFAP/decorin, which was used to generate RAd/GFAP/decorin, a blunted EcoRI human decorin fragment (1780 bp) was cloned into the blunted BamHI site of the pDE1/GFAP. 36 A blunted EcoRI human decorin fragment (1780 bp) was cloned into the blunted BamHI site of pAL119 (pMV35 in Shering et al 40 ) to create the shuttle vector pAL119/decorin which was used to generate RAd/hCMV/decorin.…”

“…21,40 To create the shuttle vector pDE1/GFAP/decorin, which was used to generate RAd/GFAP/decorin, a blunted EcoRI human decorin fragment (1780 bp) was cloned into the blunted BamHI site of the pDE1/GFAP. 36 A blunted EcoRI human decorin fragment (1780 bp) was cloned into the blunted BamHI site of pAL119 (pMV35 in Shering et al 40 ) to create the shuttle vector pAL119/decorin which was used to generate RAd/hCMV/decorin. Recombinant adenoviruses encoding the decorin gene were then generated by homologous recombination in 293 cells following cotransfection of the shuttle vector and pJM17 plasmid (Microbix biosystems, Inc.).…”

Immunological thresholds in neurological gene therapy: highly efficient elimination of transduced cells might be related to the specific formation of immunological synapses between T cells and virus-infected brain cells

“…This promoter is influenced by several regulatory mechanisms, including pro-inflammatory cytokines such as IFN-γ (Shering et al, 1997). Our experiments indicate that the immune system downregulates expression form several promoters, including viral, housekeeping and cell type-specific promoters.…”

Adenovirus-mediated expression of HSV1-TK or Fas ligand induces cell death in primary human glioma-derived cell cultures that are resistant to the chemotherapeutic agent CCNU

“…The construction of RAds expressing Escherichia coligalactosidase (RAd35 ), 24,25 HSV1 -TK ( RAd128 ) , 15,26 and mFasL ( RAdhCMV-mFasL ), 27,28 all under the control of the major immediate early human cytomegalovirus promoter ( mIE -hCMV ) , has been described in detail previously. The RAds were grown and purified as previously described.…”

“…X -Gal staining of the cultures was performed as previously described. 25 For each well, 10 fields were counted at Â10 magnification for either positive (blue cells ) or negative expression.…”