PMID- 15078955
OWN - NLM
STAT- MEDLINE
DCOM- 20040601
LR  - 20240411
IS  - 0022-538X (Print)
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Linking)
VI  - 78
IP  - 9
DP  - 2004 May
TI  - Use of amplicon-6 vectors derived from human herpesvirus 6 for efficient 
      expression of membrane-associated and -secreted proteins in T cells.
PG  - 4730-43
AB  - The composite amplicon-6 vectors, which are derived from human herpesvirus 6 
      (HHV-6), can target hematopoietic cells. In the presence of the respective helper 
      viruses, the amplicons are replicated by the rolling circle mechanism, yielding 
      defective genomes of overall size 135 to 150 kb, composed of multiple repeats of 
      units, containing the viral DNA replication origin, packaging signals, and the 
      selected transgene(s). We report the use of amplicon-6 vectors designed for 
      transgene expression in T cells. The selected transgenes included the green 
      fluorescent protein marker, the herpes simplex virus type 1 glycoprotein D (gD), 
      and the gD gene deleted in the transmembrane region (gDsec). The vectors were 
      tested after electroporation and passage in T cells with or without helper HHV-6A 
      superinfections. The results were as follows. (i)The vectors could be passaged 
      both as cell-associated and as cell-free secreted virions infectious to new 
      cells. (ii)The intact gD accumulated at the cell surface, whereas the gDsec was 
      dispersed at internal locations of the cells or was secreted into the medium. 
      (iii)Analyses of amplicon-6-gD expression by flow cytometry have shown 
      significant expression in cultures with reiterated amplicons and helper viruses. 
      The vector has spread to >60% of the cells, and the efficiency of expression per 
      cell increased 15-fold, most likely due to the presence of concatemeric amplicon 
      repeats. Current studies are designed to test whether amplicon-6 vectors can be 
      used for gene therapy in lymphocytes and whether amplicon-6 vectors expressed in 
      T cells and dendritic cells can induce strong cellular and humoral immune 
      responses.
FAU - Borenstein, Ronen
AU  - Borenstein R
AD  - The S. Daniel Abraham Institute of Molecular Virology and Department of Cell 
      Research and Immunology, Tel Aviv University, Tel Aviv 361390, Israel.
FAU - Singer, Oded
AU  - Singer O
FAU - Moseri, Adi
AU  - Moseri A
FAU - Frenkel, Niza
AU  - Frenkel N
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (Luminescent Proteins)
RN  - 0 (Membrane Proteins)
RN  - 0 (Proteins)
RN  - 0 (Viral Envelope Proteins)
RN  - 0 (glycoprotein D, Human herpesvirus 1)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Gene Deletion
MH  - Gene Expression
MH  - *Genetic Vectors
MH  - Green Fluorescent Proteins
MH  - Helper Viruses/*genetics
MH  - Herpesvirus 6, Human/*genetics
MH  - Humans
MH  - Jurkat Cells
MH  - Luminescent Proteins/genetics/metabolism
MH  - Membrane Proteins/genetics/metabolism
MH  - Proteins/genetics/metabolism
MH  - T-Lymphocytes/*metabolism
MH  - Transgenes
MH  - Viral Envelope Proteins/genetics/*metabolism
PMC - PMC387683
EDAT- 2004/04/14 05:00
MHDA- 2004/06/02 05:00
PMCR- 2004/05/01
CRDT- 2004/04/14 05:00
PHST- 2004/04/14 05:00 [pubmed]
PHST- 2004/06/02 05:00 [medline]
PHST- 2004/04/14 05:00 [entrez]
PHST- 2004/05/01 00:00 [pmc-release]
AID - 1856 [pii]
AID - 10.1128/jvi.78.9.4730-4743.2004 [doi]
PST - ppublish
SO  - J Virol. 2004 May;78(9):4730-43. doi: 10.1128/jvi.78.9.4730-4743.2004.